A method for treating steel works dust in order to recuperate elements capable of being upgraded. The method comprises attrition in water followed by hydraulic grading of the resulting load. The method is characterized in that is further comprises: a washing step to separate the water soluble saline fractions of the insoluble oxides; hot treatment to eliminate metals in the form of free oxides such as zinc and lead; treatment by heating at a temperature ranging between 240 and 800xc2x0 C.; treatment with sulphuric acid having a concentration between 5 and 8%.

Field of the Invention
The invention relates to the technical field of the treatment of dust from steelworks, notably electric steelworks.
According to a first general object, the invention discloses a method and an installation for treating said dust allowing elimination of the heavy metals it contains.
According to a second general object, the invention discloses a method and an installation for treating said dust allowing maximum economic recovery of the most abundant fraction of this dust.
The term heavy metal, which has come into use in many legislations, relates to metals which are generally:
non-degradable over time;
toxic to living systems at very low concentrations;
inclined to accumulate in living organisms and to become concentrated in the course of transfers of materials in food chains.
Steelworks dust, for example issuing from remelting in the electric furnace of scrap iron for the production of steel, contains such heavy metals such as zinc, cadmium and lead, for example.
Dust production is between 15 and 22 kg per tonne of liquid steel.
As a guide, steel production, in Europe, was of the order of over 500,000 tonnes per year at the beginning of the 1990s.
Consequently, there is a considerable need for effective treatment of this dust, for the sake of protecting the environment notably.
A number of methods have been envisaged for the treatment of this dust, the methods varying depending on the composition of said dust.
Two main classes of dust are distinguished, depending on the nature of the steels produced, carbon steels and stainless alloyed steels.
In the case of carbon steels, zinc (17 to 29%) is present in the dust in two forms: zinc oxide, ZnO, and zinc ferrite, ZnFe2O4, while the lead (4 to 5%) is in oxide form (PbO) The cadmium contents are lower, of the order of around 800 ppm.
Unlike zinc oxide, zinc ferrites are not easily soluble during hydrometallurgical treatments for releasing the zinc.
For alloyed and stainless steels, the quantity of dust generated in Europe was around 50,000 tonnes per year during the 1990s.
Various procedures and processes, many still at the experimental laboratory stage, have been devised for the treatment of dust from steelworks, notably electric steelworks.
The BUS (BERZELIUS UMWELT) procedure proposes a treatment of dust from steelworks for carbon steels combining two processes: the WAELZ process and the ISP (IMPERIAL SMELTING PROCESS). This procedure uses a conventional pyrometallurgical process and condensation of the zinc vapours by means of liquid lead spray condensers.
The FLAME REACTOR process, used for the treatment of dust from alloyed and stainless steels, consists essentially of vertical flame smelting/reduction cyclone separation. The waste products, rich in zinc and lead, are converted into slag to be resmelted and into oxide to be recovered.
The PLASMADUST process has been used since the 1980s for the treatment of electric steelworks dust rich in zinc and lead. The treatment furnace is of the tank furnace type, whose energy is provided by blown arc plasma torches. The energy of the plasma is used to heat the reagents, and to provide the heat necessary for the endothermic reduction reactions. The plasma-producing gas is introduced into the furnace by water-cooled copper nozzles. The powdery charge composed of dust, flux and coal is injected into the nozzles and mixed with the plasma-producing gas. The fumes charged with metallic vapours leave the furnace at a temperature of around 1150xc2x0 C.
When the PLASMADUST process is used for dust from alloyed and stainless steels, the zinc content in the fumes is generally too small for condensation of the zinc to be efficient, the zinc in this case being retrieved from the sludge issuing from the washers.
The PLASMINOX process uses a treatment by means of hot cathode plasma torches operating by transferred arc at D.C. and installed vertically on the furnace. In the furnace, the metals presentxe2x80x94chromium, nickel, molybdenum and ironxe2x80x94are separated from the dust by melting and the ferroalloy which results therefrom is poured into moulds for subsequent recharging at the dust-producing steelworks. The scorias obtained are, in accordance with the standards, inert. The fumes escaping from the furnace are cooled and have their dust removed. The secondary dust obtained contains volatile metals Zn, Pb which are in the form of oxides and are not recovered.
The modified ZINCEX process produces zinc by hydrometallurgical means, this process having the following successive steps:
atmospheric leaching of the dust by a dilute sulphuric acid solution;
purification of the leaching solution by precipitation of the iron and aluminium;
selective extraction of the zinc by D2EHPA;
stripping of the zinc from the organic phase by the highly acid electrolytic solution;
electrodeposition of the zinc on to aluminium cathodes.
The ZINCEX process is relatively complex in its implementation since it requires organic solvents combined with sulphuric acid. Furthermore, this process requires large quantities of wash water to avoid the formation of SO2 from the oxides which are recycled in the furnace. This ZINCEX process is not easily exportable, owing to the above-mentioned problems and since it necessitates too heavy investments.
The GLASSIFICATION process comprises a step of mixing the steelworks dust with other steelwork waste products and constituents of glass. The mixture is melted in a submerged arc furnace. The metals vaporized during the melting condense in the upper part of the furnace before being trapped in the melt. The glass obtained can be put in the form of granules for the manufacture of glass tiles or grit.
In total, three main ways of recycling have been envisaged for recycling dust from electric steelworks.
A first way consists of performing reductions at various temperatures in accordance with the processes for vaporizing the heavy metals and re-injecting the ferrous fraction into the steel production furnaces:
the document EP 336 923 proposes treating steelworks dust in an iron smelting furnace after conversion into pellets;
the document EP 441 052 proposes a thermal treatment between 1200 and 1700xc2x0 C. by the addition of reducing agents;
the document WO 91/12 210 describes a method of treating steelworks dust in an iron reduction furnace, the heavy metals being retrieved by condensation from the hot gases;
the document EP 453 151 describes a method of treating dust in the form of pellets by an agent selectively reducing iron oxide;
the document FR 2 666 592 describes a device for extracting volatile metals acting by oxidation;
the document WO 93/69 619 describes an arc furnace specially designed for the reduction of dust containing oxides;
the document EP 551 992 describes a method of retrieving recoverable metals from steelworks dust by reduction and vaporization of the recoverable metals;
the document FR 2 373 612 describes a method of extracting zinc contained in steelworks dust, by selective oxidation under heat.
A second way consists of directing the dust towards a use where it would be xe2x80x9cmade inertxe2x80x9d by various means with a view to use as construction or filler materials:
the document EP 402 746 proposes recycling by incorporation in a clay for a mine packing material;
the document WO 91/12 210 proposes the use of steelworks dust for the treatment of sewage with the addition of a flocculating agent;
the document FR 2 689 881 describes a method of manufacturing bricks whose properties are improved by the addition of electric steelworks dust first calcined at the vaporization temperature of heavy metals contained in said dust;
the document FR 2 700 161 describes a mixture for a road surfacing comprising 2 to 6% by weight of electric steelworks filter dust.
Pyrometallurgical processes, with or without electricity, allow the recovery of oxides of zinc, of iron or nickel chromium alloy or of liquid zinc most often by condensation.
A third way, hydrometallurgical, has also been envisaged.
Various media such as sulphuric acid, hydrochloric acid, soda and ammonia have been tried out.
The document FR 2 716 895 describes a method of treating electric steelworks dust with or without prior reduction, or electrolytic galvanizing dust. Said method comprises an ammoniacal attack by an ammoniacal buffer solution, then a desorption of the ammonia by air flow cleaning.
Direct attacks in a sulphuric or hydrochloric medium are not economically viable, since the volumes in terms of iron hydroxide or salts involved are enormous.
These residues are moreover still contaminated by metallic impurities such as lead and cadmium.
Alkaline leaching has been proposed.
The document FR 2 535 736 describes a method of treating dust containing zinc, issuing from electric steelwork furnaces, in which the dust is subjected to a basic leaching. The basic rejects resulting from the different washing phases and containing zinc are neutralized by means of the acid rejects coming from galvanization operations and also containing zinc.
The document FR 2 501 141 describes a basic leaching of zinc-bearing dust by means of ammonium chloride, the powder obtained being agglomerated and introduced into a smelting furnace.
By the leaching technique, only the elements in the form of free oxides (mainly zinc and lead) are dissolved. The mixed oxides of non-ferrous metals/ferrous oxides are not affected. Their management is therefore problematic, including re-injection into the steelworks furnace which leads, in the end, to a problem of re-concentration of impurities.
The methods combining liquid ammonia with ammonium salts (notably the ENGITEC process described in European patent application EP-93 00018.5) have drawbacks in the handling of chlorides for the retrieval of minority elements (zinc, lead and cadmium) and the recovery of iron oxides. The high chloride content prevents recycling of these phases in the furnaces owing to the problems related to chlorine. Their washing would require considerable volumes of water.
It can therefore be thought that this type of method has a maximum efficiency between 25 and 30%, proportional to the concentration in the non-ferrous phase. This method therefore generates a waste product whose management remains as problematic as that of the untreated dust.
The document FR 2 737 503 describes a method of preparing mineral pigments, and the mineral pigments thus obtained, and an installation for the implementation of such a method. This document, coming from the applicant, describes a method having the following steps:
separation of the dust into two fractions, one fraction comprising magnetic elements and one fraction comprising non-magnetic elements;
basic leaching of the non-magnetic fraction in order to dissolve the zinc fraction which would not be bound up in spinel form, and the silica, lead, and a fraction of the manganese;
rinsing until neutralized and separation of the charge thus obtained;
calcination at a temperature between 450 and 650xc2x0 C. of the charge thus obtained;
treatment of the calcined charge with sulphuric acid in the presence of a catalyst for solubilizing the iron oxides formed during the calcination step and the heavy metals other than zinc;
collection of the mineral pigments;
use of the solutions issuing from the rinsing and the sulphuric acid treatment for precipitating other pigments.
The present invention proposes a method of neutralizing the free heavy metals contained in the dust from steelworks, notably electric steelworks, said method also allowing the recovery of the most abundant fraction of said dust.
The method according to the invention makes it possible to treat indiscriminately the so-called acid dust and the so-called basic dust.
The method according to the invention makes it possible to geographically dissociate pre-treatment of the dust at the collection site and chemical treatment and recovery from said dust.
The method according to the invention is particularly adapted to the treatment of dust from carbon steels and tool steels.
The invention relates, according to a first aspect, to a method of treating steelworks dust with a view to retrieval of the recoverable elements, said method comprising an attrition in water followed by a water grading of the charge thus obtained, said method being characterised in that it also comprises:
washing of the charge collected at the overflow issuing from the water grading, said charge being charged with dissolved heavy metals and salts, with a view to separating the water-soluble saline fractions from the insoluble oxides,
treatment under heat of the whole of the charge, washed in the previous washing step without prior magnetic separation, with a view to eliminating the metals in the form of free oxides such as zinc and lead,
treatment by thermal means of the charge obtained after treatment under heat at a temperature between 240 and 800xc2x0 C.
The method also has the following aspects, possibly combined.
A step of treatment with sulphuric acid with a concentration of between 5 and 8% is carried out after the treatment by thermal means.
The treatment under heat performed after the washing step is, according to a first variant, carried out in an acid medium. Said acid medium is, in one embodiment, a sulphuric acid solution at a concentration of between 8 and 18% and a temperature of between 40 and 95xc2x0 C.
The treatment under heat is followed by an oxidation and then a separation of the iron in the form of an insoluble oxide precipitate.
The zinc is separated by means of an acid electrolysis.
The treatment under heat performed after the washing step is, according to a second variant, carried out in an alkaline medium.
According to one embodiment, the washing comprises treatment with a sulphur derivative chosen from amongst the group of heterocyclic compounds such as trimethyl triazine or 2,5 dimercapto thiadiazole.
The alkaline treatment is performed in a concentrated soda solution under heat in the presence of an oxidizing agent.
The concentration of the soda solution is between 240 and 400 g/l.
The temperature of the alkaline treatment is between 50 and 110xc2x0 C.
The oxidizing agent is chosen from amongst the compounds which are oxidizing in an alkaline medium such as hydrogen peroxide.
The hydrogen peroxide concentration is between 10 and 40%, and more particularly between 30 and 35%. The lead contained in the alkaline solution is eliminated by means of hydrogen peroxide with a concentration equal to at least 35% by weight.
The treatment by thermal means is performed between 400 and 650xc2x0 C.
The treatment with sulphuric acid is carried out in the presence of an injection of air in order to oxidize the divalent iron into trivalent iron.
The treatment with sulphuric acid is carried out under heat at a temperature between 40 and 80xc2x0 C.
The solution issuing from the acid attack is adjusted to a pH between 5.4 and 10.
The solution issuing from the acid attack is adjusted to a temperature between 25 and 80xc2x0 C.
The solution issuing from the acid treatment is purified by means of iron or zinc powders.
The method also comprises a two-stage electrolysis of the solution issuing from the alkaline treatment.
Said first electrolysis uses a cathode of graphite in grains contained in a basket in contact with a titanium grid.
The basket is made of polypropylene.
The anode is made of titanium containing ruthenium or iridium.
The second electrolysis comprises an anodic re-dissolution of the graphite cathodes in a sodium sulphate solution and a cathodic re-deposition of the zinc on an iron or steel electrode.
The second electrolysis is carried out in an electrolyser whose pH is adjusted to a value close to 5.
Other objects and advantages of the invention will emerge in the course of the following description of embodiments, said description being given with reference to the accompanying figures in which:

Method for treating steel works dust by wet process


There is provided a liquid discharging apparatus including: an original drive signal generation unit which generates an original drive signal; a signal modulation unit which modulates the original drive signal to generate a modulation signal; a first signal amplifying unit which amplifies the modulation signal to generate a first amplified modulation signal; a second signal amplifying unit which amplifies the modulation signal to generate a second amplified modulation signal; an amplification control unit which controls operations of the first signal amplifying unit and the second signal amplifying unit; a signal conversion unit which converts the first amplified modulation signal and the second amplified modulation signal into a drive signal; a plurality of piezoelectric elements which is deformed by the drive signal; a plurality of cavities which expands or contracts in accordance with the deformation of the plurality of piezoelectric elements; a plurality of nozzles which communicates with each of cavities.

1. Technical Field
The present invention relates to a liquid ejecting apparatus which ejects liquid by applying a drive signal to an actuator, a control method thereof, and a program, and is preferable for a liquid ejection type printing apparatus which is designed to print predetermined characters, images, and the like by ejecting minute liquid droplets from nozzles of a liquid ejecting head and forming fine particles (dots) on a print medium.
2. Related Art
As one example of liquid discharging apparatuses, an ink jet printer which discharges ink (liquid) from nozzles provided at a head to a recording medium has been known. Generally, a serial head scheme in which a nozzle array including multiple nozzles aligned in a predetermined direction is formed at a head and an image with a nozzle array width is printed by the head discharging ink while relatively moving in an intersecting direction between a scanning direction of the head and a transport direction of a recording medium, for example, a line head scheme in which nozzles are arranged in an array in a direction intersecting with a transport direction of a print medium and an image is printed when the print medium passes below the nozzles, and the like as disclosed in JP-A-2011-5733, and the like have been known.
Methods of ejecting liquid from nozzles of a liquid ejecting head include an electrostatic scheme, a piezoelectric scheme, and a film boiling liquid ejecting scheme. In a case of the piezoelectric scheme, for example, if a drive signal is applied to a piezoelectric element as an actuator, then a vibration plate in a cavity is displaced, pressure change occurs in the cavity, and liquid is ejected from nozzles by the pressure change. In a case of a serial head scheme high-speed printer in which liquid is ejected at a high speed by causing a liquid ejecting head to perform scanning at a high speed and driving a large number of piezoelectric elements in short time, and in a case of a line head scheme liquid ejection type printing apparatus or the like in which liquid is ejected at the same time from a plurality of nozzles by simultaneously driving a plurality of piezoelectric elements, it is necessary to drive a large number of piezoelectric elements, and burden applied on a drive circuit per unit time is significantly large. Therefore, it is generally difficult to generate the drive signal by using the same configuration as that of a serial head scheme ink jet printer in the related art, which has been provided in the consumer market, without any change.
Thus, a method of using a plurality of Digital-to-Analog Converters (DACs) and a plurality of amplifying circuits (hereinafter, also referred to as amplifiers) to generate a plurality of drive signals and equally dividing the number of nozzles to be supported by one drive signal can be considered. However, in a case of providing the plurality of DACs and amplifiers, errors of the respective DACs and errors of the respective amplifiers increase in a multiplied manner due to the combination thereof. If errors of the driven piezoelectric elements are further taken into consideration, errors as a whole further increase. As a result, it is difficult to perform overall control, and quality of a material produced by the liquid ejection type printing apparatus may deteriorate.
Here, it is preferable to generate the drive signal by using a single DAC and a single amplifier in order to minimize an influence of the errors. However, there is a limitation in power supply of the amplifier (for example, there is a limitation in allowable current of a circuit in an output stage). Therefore, it is not possible to appropriately drive a large number of piezoelectric elements and quality of the produced material deteriorates in the case of the piezoelectric scheme, for example, and therefore, such a configuration is not realistic.

Liquid discharging apparatus, control method thereof, and program


Disclosed is a control system for use with one or more building power circuits, the control system including a control device delimited by a control device housing, the control device being configured to receive power from the building power circuits and provide power to at least one power output, and a receiver configured to receive radio frequency signals, the receiver being disposed remote of the control device housing and configured for removable wired connection with the control device.

In the interest of energy conservation and given the proliferation of portable plug-in electrical devices used in offices, there is a growing need for control of convenience power receptacles and/or receptacle circuits deployed in offices. This especially applies to power receptacles deployed in association with portable and modular office furniture. The need is further elevated by energy conservation legislation that requires the implementation of such control in buildings. However, notwithstanding the need to implement switching control of office power receptacle circuits, it is equally desirable, and at the same time necessary, to provide unswitched convenience power in offices for devices that must remain energized on a 24-hour schedule. These devices include networked personal computers and the like.
The current art includes circuit controllers that are wired in association with building wiring and modular wiring to provide on/off control of circuits that may include one or more plug load or convenience power receptacles controlled in response to signals received at the controller via a digital data link and/or from one or more sensors connected to the controller. However, a disadvantage of such controllers is the lack of a wireless (i.e. radio frequency) input and thus the need for access to a wired data or signal link when interfaced with remote sensors, sensor networks, and/or building energy management systems.
In addition, the current art currently includes circuit controllers that are wired in association with power wiring so as to provide on/off control of circuits inclusive of one or more convenience power receptacles in response to signals received from one or more sensors connected directly to the controller and via radio frequency transmissions from remote sensors, sensor networks and/or building energy management systems received via an imbedded radio receiver. However, one disadvantage of such controllers with embedded radio receivers is their inability to detect or "hear," and reliably respond to, wireless control commands when said controllers and receivers are disposed in the context of modular, plug-and-play multi-circuit power systems such as those deployed in modular furniture and the like.
The current art further includes portable controllers powered by a cord and plug, which provide dimming control of one or more controllable luminaires via plug-and-play connectors and plug-and-play cabling, wherein the diming occurs in response to sensors connected to the controller via plug-and-play connections and via radio frequency transmissions from remote sensors, sensor networks, and/or building energy management systems received via an imbedded radio receiver. However, one disadvantage of such controllers is the lack off an ability to desirably control (i.e. energize and de-energize) circuits that may include one or more convenience power receptacles.
The current art still further includes portable controllers powered by a cord and plug, which provide switching control of one or more portable electrical appliances via plug-and-play connections, the control occurring in response to radio frequency transmissions from remote sensors, sensor networks, and/or building energy management systems received via an imbedded radio receiver. However, such controllers lack the ability to inherently establish a desirable mix of controlled convenience power receptacles and uncontrolled convenience power receptacles associated with portable office furniture systems.

Control system for use with one or more building power circuits


An engine is disclosed having a water cooling system allowing efficient cooling of the exhaust valves to prevent temperature gradients from building in the engine. Water is therefore pumped through the engine though first and second water cooling cores which discharge through the head. A water manifold is positioned over the discharge opening and includes couplings for the radiator supply, radiator return, water pump supply, oil cooler supply and oil cooler return. The engine has separate chambers to isolate the pistons and cylinders, and reed valves cover the chambers and allow the blow-by gases and oil to enter the oil pan during the power stroke of the engine cycle.

The present application relates generally to internal combustion engines, and particularly water cooled engines.
Multiple different engine types are known, for example, multiple fuel types are available, and multiple different sized engines are available, together with different numbers of cylinders. Engines may also be 2 or 4 stroke, and be positioned at multiple different orientations, for example the piston(s) may be oriented vertically, horizontally, or at any other possible orientation. It is also known to cool the engines by either air or water circulation. The subject disclosure is applicable to all types of such engines.


The movement detector having six degrees of freedom comprises a support on which three position sensors are arranged according to three orthogonal axes. Each sensor comprises a rigid body (3), a deformable element (4) which is electrically conductive, and four conductive areas (6b) which are disposed on rigid walls (3). The deformable element (4) is balanced around the centre part thereof and has a rest position and four active positions wherein it is temporarily in contact with two of the conductive areas (6b) of the rigid body (3). The deformable element (4) of a sensor moves from a rest position to one of the active positions in response to a predetermined directional or axial translation or votation.

The invention relates to a movement detector with six degrees of freedom comprising a support on which three position sensors are arranged disposed along three axes, each sensor comprising a rigid body, conducting areas disposed on the rigid body and an electrically conducting deformable element, presenting a rest position wherein it is isolated from the conducting areas, and moving from the rest position to an active position in response to a high-speed movement of predetermined direction and orientation.
The invention also relates to a method for production of a sensor.

Movement detector having six degrees of freedom with three position sensors and method for the production of a sensor


The present application is directed to the processing of a biological sample into its constituent components for use in ART and includes introducing a sample into a first volume disposed adjacent a second volume including buffer solution, wherein the first and second volumes are adapted for fluid communication therebetween, selectively separating the first volume from the second volume with a movable closure member disposed therebetween, wherein the step of selectively separating the first volume from the second volume includes moving the closure member so that a fluid communication aperture is formed by one or a combination of the closure member or the closure member in combination with the first and second volumes to allow fluid communication between the first volume and the second volume such that motile cells migrate from the sample in the first volume to the buffer solution in the second volume.

Throughout this specification the use of the word "inventor" in singular form may be taken as reference to one (singular) inventor or more than one (plural) inventor of the present invention.
It is to be appreciated that any discussion of documents, devices, acts or knowledge in this specification is included to explain the context of the present invention. Further, the discussion throughout this specification comes about due to the realisation of the inventor and/or the identification of certain related art problems by the inventor. Moreover, any discussion of material such as documents, devices, acts or knowledge in this specification is included to explain the context of the invention in terms of the inventor's knowledge and experience and, accordingly, any such discussion should not be taken as an admission that any of the material forms part of the prior art base or the common general knowledge in the relevant art in Australia, or elsewhere, on or before the priority date of the disclosure and claims herein.
In Assisted Reproductive Technologies (ART), including clinical processes such as, for example, in vitro fertilisation (IVF), intracytoplasmic sperm injection (ICSI) and intrauterine insemination (IUI), semen samples donated by the intended father are required to be processed to make them suitable for use. The main purpose of this processing is the removal of seminal plasma constituents that have negative impact on sperm viability and function. In vivo the seminal plasma is ordinarily removed during the sperm passage through the female reproductive tract. In addition to separating the plasma, the processing of semen samples aims for concentration and enrichment of the most viable and functional sperm population from the original semen sample, manifested by their motility.
Currently most widely used sperm processing methods include density gradient centrifugation (DGC) and so-called swim-up methods, although some other methods have also recently entered the markets.
With reference to FIG. 1, in the DGC method, a neat semen sample is placed into a centrifuge tube on top of two layers of colloidal suspension of silica particles in buffer. The upper phase or top layer consisting of lower concentration of particles, for example 45%. The lower phase or bottom layer consisting of higher particle concentration, for example 90%. The tube is then centrifuged, typically for 20 minutes at 1,600 rpm, allowing sperm to move through the 'upper phase' and 'lower phase' layers according to their properties, resulting in viable motile sperm ending up at the bottom of the tube, dead cells and debris within and/or between the silica layers, and seminal plasma remaining at the top of the tube. Several commercial silica compounds are available for this purpose, including for example Gems™ Sperm Wash Gradient Set (Genea Biomedx), SupraSperm™ (Origio), SpermGrad™ (Vitrolife) and many others. Following this step the motile sperm needs to be washed with sperm medium, sperm buffer or even fertilization medium via centrifugation at least once more to remove remnants of the silica compound.
DGC has advantages in that it provides for separation of motile sperm, removal of immotile sperm, debris and seminal plasma and it can also accommodate variable sample volumes for processing. However, DGC also has disadvantages in as much as there can be limited separation of morphologically abnormal sperm. Other disadvantages include a risk of user error in handling, variability of results, exposure to potentially harmful substances in the silica reagent, unwarranted effects of centrifugal forces, long processing times and the requirement for moving sample across several containers during different steps of the process. Nonetheless, DGC is currently the most common industry practice.
With reference to FIG. 2, in the swim-up method, a liquefied semen sample comprising neat semen is carefully placed at the bottom of a centrifuge tube containing sperm medium or buffer and left to sit in an incubator at a temperature of 37° for about an hour, during which time the motile sperm ie viable spermatozoa literally swims up the tube. The top layer of the media containing the motile sperm is then collected, leaving seminal plasma, dead sperm and debris behind. This collected portion is often centrifuged once more to concentrate the preparation, as sperm is usually needed in high concentration for the final processing step.
The swim-up method for sperm processing has advantages of being simple because it does not require complicated consumables or reagents. Further, variable sample volumes may be processed. However, there are disadvantages, for example, in that the sample cannot be left in the tube for extended periods due to its unfettered exposure to seminal plasma, prolonged exposure to which after ejaculation is known to diminish fertilizing capacity of human spermatozoa. Furthermore, it can be prone to the risk of user error, it has long processing time, it requires sample transfer across several containers during the different steps of the process and when the centrifugation step occurs sperm is exposed to the unwarranted effects of centrifugal forces. It is also a labour-intensive manual process. Nonetheless, the swim-up method is also current industry practice.
The problems encountered with both the DGC and swim-up methods relate to complexity and length of the process. Both require movement of the semen sample from the initial sample container through several different containers (and sub-containers for sub-samples) before reaching the IVF/ICSI dish or IUI catheter. Each such movement increases the risk of sample damage, loss of traceability, contamination, and requires a manual handling step as well as an associated double-witnessing/automatic witnessing step to ensure correct ID traceability throughout the process. The duration of the required procedures varies from 40 minutes to over an hour, adding complexity to the laboratory work flows as well as tying up laboratory equipment and space. The nature of the processes alternating between hands-on activities and waiting periods complicates the workflows even further.
Most laboratories adhere to stringent requirements of handling only one sperm sample at a time in order to reduce the risk of mix-up, and because of this, bigger laboratories have to maintain several sperm processing stations, each with their own devoted equipment such as laminar flow hoods, pipette sets and sometimes also a centrifuge. The need for manual labour is also high due to many consecutive steps requiring not only the time resources of the operator, but in many cases also time resources of the second witness to ensure sample safety and traceability. This in turn has flow-on effects to other ART operations by requiring the witness to interrupt their tasks, thus increasing the overall risk of operator error.
The large component of manual handling also makes the process susceptible for human error and variation resulting from different skill levels of the operators. The complexity of the process also calls for a large number of reagents and consumables to be used for the process, which involves further inherent stock managing, consumable and media risks and cost implications.
In addition, it has been shown that the DGC method may cause DNA damage in sperm, which in turn may have impact on not only sperm viability and functionality, but also on embryo development and possibly resulting pregnancy given that sperm DNA represents 50% of the genome of the embryo.
Some recent sperm processing systems rely on standardisation of some aspects of the previous methods, such as the swim-up-based system Seaforia™ with specifically designed consumables and a warming station, or likewise the swim-up based RI MSC™ sperm separation tube. The Seaforia™ system (described at the following Internet resource link http://www.lotusbio.com/index.html?page=37) has the advantages of a controlled environment, ease of use and utilises standardised volumes but disadvantageously, it takes a long time, requires sample movement across several containers and requires a separate final output collection and analysis just like DGC and swim-up method do. As a new market entrant, information about clinical outcomes is limited. RI MSC is a simple procedure like swim-up but has drawbacks of having no active gating, is prone to user error, has a long process time, requires sample movement across several containers and also requires a separate final output collection and analysis. RI MSC has been in the market for several years.
Some systems use microfluidic designs that allow motile sperm to navigate through a microchannel and porous membrane such as; FERTILE® which is described at http://koekbiotech.com/fertile/, FERTILE PLUS® which is described at http://koekbiotech.com/fertile-plus/. There is also the FERTILE Ultimate® system, described at http://koekbiotech.com/fertile-ultimate/. These are all based on sperm travelling through a microfluidic channel from a place of deposition to a place of collection. The FERTILE® technique is simple and does not expose sperm to DGC reagents or centrifugal forces linked with causing DNA damage, but it has disadvantages in that it takes a long time, is impacted by the external conditions such as temperature and is costly. In addition, the FERTILE® system especially processes very low volumes and all systems require a separate final output collection and analysis. As the FERTILE® systems are new entrants to the market, information about clinical outcomes is limited.
Some known systems use filaments such as the Zech Selector™ instrumentation, described in US patent publication No. 2013/0164838. Likewise, Zech Selector™ is simple and does not expose sperm to factors promoting DNA damage but it has disadvantages in that it also takes a long time, requires fixed fluid volumes, and also requires a separate final output collection and analysis. Being a new market entrant, information about clinical outcomes is also limited.
Other known systems use a multitude of microfluidic channels, such as the QSperm system, which however has not reached commercial launch as yet. It is described at http://marsinnovation.koazoa.com/wp/tag/qsperm/.
Further known systems involve electrophoresis-based technologies relying on sperm selection in an electronic current based on their membrane potential. The expected advantages of these methods include that sperm are not exposed to known factors promoting DNA damage, and the technique may be amenable to be undertaken over a short time. However, there are unknown effects of electrophoresis reagents and processes upon sperm and these require complicated instruments, consumables and reagents. Also, none of these systems have reached commercial launch yet and information about clinical outcomes is not available or is very limited, being experimental only.
A number of other specific prior art systems are noted below.
PCT Publication No. WO 2012/162181 in the name of The Brigham And Women's Hospital, Inc. entitled, "Analysis and Sorting of Motile Cells" discloses a method for sorting motile cells including introducing an initial population of motile cells into an inlet port of a microfluidic channel, the initial population of motile cells having a first average motility; incubating the population of motile cells in the microfluidic channel; and collecting a sorted population of motile cells at an outlet port of the microfluidic channel. The sorted population of motile cells has a second average motility higher than the first average motility.
PCT Publication No. WO 2013/129947 in the name of Auckland Uniservices Limited entitled "Method and Apparatus for the Isolation of Motile Sperm" discloses the use of 'edge-trained' motile sperm and is directed to a method and system for separating motile and nonmotile sperm which comprises delivering a fluid containing sperm into a microvolume at least partially defined by a wall which includes a wall termination or a change in angle away from the microvolume that is noted as being a wall termination, and allowing at least some motile sperm to move along said wall and to exit the microvolume by changing direction away from the microvolume at or near the wall termination, to or towards collection of motile sperm. In broad terms in another aspect the invention comprises a method and apparatus for separating motile and non-motile sperm which comprises delivering a fluid containing the sperm into a microchannel comprising a wall which includes a wall termination, allowing motile sperm to move in substantially no-flow conditions in the microchannel along the wall and to exit the microchannel by changing direction away from the microchannel at or near the wall termination, to or towards a collection reservoir, and recovery of fluid containing the sorted sperm from the collection reservoir.
U.S. Pat. No. 5,028,526 in the name of Alice Deutsch entitled "Method for Semen Analysis" discloses a method for the separation of seminal plasma from semen by means of a membrane. The invention also includes a method for the determination of enzymes such as acqrosin and other components of semen.
United States Patent Publication No. US 2006/0270021 to Shuichi Takayama, et. al entitled "Integrated Microfluidic Sperm Isolation and Insemination Device" discloses an integrated microfluidic sperm isolation and oocyte insemination device, which provides the opportunity to perform in vitro insemination with motility enhanced sperm samples and with minimal manipulation of fragile oocytes. Sperm sorting is performed in a common sort channel wherein more mobile sperm swim across the interface between co-laminar flows of semen and media fluid.
United States Patent Publication No. US 2010/0291535 to Da-Jeng Yao, et al. entitled "Method Using Microfluidic Chip to Sort High Motility Sperm" discloses a method using a microfluidic chip to sort high motility sperm. Sperm and a medium are respectively injected into a microchannel of a microfluidic chip via several inlets. Owing to the characteristic of microfluidics, the sperm and the medium form a sperm laminar flow and medium laminar flow in the microchannels; the sperm laminar flow and the medium laminar flow are parallel to each other. The higher motile sperm may pass through at least one laminar flow within a limited time, whereby different motility levels of sperm can be respectively collected from different outlets.
United States Patent Publication No. US 2012/0118740 to The Regents of The University of California entitled "Methods and Devices for Sorting Cells and Other Biological Particulates" discloses an optical pattern-driven light induced dielectrophoresis (DEP) apparatus and separation methods are described which provide for the manipulation of particles or cells and selection based on traits correlated with the DEP response. Embodiments of the apparatus use DEP electric field patterns in combination with microfluidic laminar flows to measure response, separate, segregate and extract particles from heterogeneous mixtures according to the relative response of the particles to one or more DEP fields without damaging living cells. The methods are particularly suited for selecting and extracting the best sperm and embryo candidates based on fitness for use with existing artificial reproduction procedures and excluding defective or non-viable gametes.
United States Patent Publication No US 2008/0187991 to The Regents of The University of Michigan, entitled "Process for Sorting Motile Particles from Less Motile Particles" discloses a technique in which motile particles are sorted from non-motile particles in a microfluidic sorting device wherein a stream of sort fluid containing motile and non-motile particles is caused to flow adjacent a media stream in non-turbulent fashion through a sort channel, during which flow motile particles cross the interface between the adjacent flow streams, entering the media stream, and forming a motile particle-depleted sort stream. The sorting devices are easily and inexpensively fabricated and have numerous uses, in particular, sorting of motile from non-motile sperm.
PCT Publication No. WO 2012/032165 to Josef Zech entitled "Method for Enrichment of DNA Strand Break-Free Spermatozoa and Reduction of Risks for Abnormalities and/or Aneuploidy" discloses a method for producing an enriched DNA strand break-free spermatozoa sample from a poor quality seminal fluid, wherein a selecting device which comprises two chambers and a bridge element is used and the method comprises placing a seminal fluid comprising at least 15% oligo-, at least 32% astheno-, and/or at least 4% teratozoospermia, based on the total amount of the spermatozoa in the seminal fluid, in the first chamber of the selecting device, filling the second chamber of the selecting device with a medium for receiving DNA strand break-free spermatozoa and connecting both chambers by a bridge element such that a fluid bridge between the first and second chamber occurs which allows the DNA strand break-free spermatozoa to move from the first to the second chamber.
United States Patent Publication No US 2009/101507 originally assigned to The University of Newcastle Research Associates Limited, entitled "Sperm Cell Separation By Electrophoresis" discloses a process for separating a sperm type from a sperm population by electrophoresis in a modification of a laboratory electric circuit for electrophoresis, which involves subjecting the sperm population to an electric potential such that a sperm type is separated from a sperm population through an ion-permeable barrier.
PCT Publication No. WO 1994/017742 to Josef Zech entitled "Device for Removing Sperm Cells from Seminal Fluid" discloses a device for in-vitro fertilization with a container for ova and a container for sperm cells, in which a capillary tube provides a liquid bridge between the containers. Preferably the liquid bridge extends between the ova container and a part which passes round its upper edge in a U-shaped fashion.
PCT Publication No. WO 2003/031564 to Josef Zech entitled "Device and Method for Selecting Locomotive Biological Species, Particularly Sperm Cells" discloses a device for selecting locomotive sperm cells, comprising a first chamber provided for accommodating a medium containing the species to be selected and a second chamber, which is separate from the first and provided for accommodating the selected species in another medium. A bridge element, which can be placed on the chambers and which has at least one flat-like channel. Said channel is formed by delimiting walls, extends from the first chamber to the second chamber when the bridge element is placed thereupon, and has an opening in the area of the first chamber and in the area of the second chamber. In addition, the channel can be filled with a medium, in which the locomotive biological species can move, whereby bringing the medium in the first chamber in contact with the medium in the second chamber.
PCT Publication No. WO 2014/006043 to Josef Zech entitled "Device for Spermatozoa Selection" discloses a first chamber configured to receive a first, seminal fluid; a second chamber configured to receive a second fluid, the second chamber being in fluid communication with the first chamber by means of at least one duct having a first opening to the first chamber and a second opening to the second chamber; and a displacement means adapted to displace at least some of the first, seminal fluid towards the first opening.
U.S. Pat. No. 6,391,654 to Genosis Limited entitled "Separation and Detection of Spermatozoa" discloses a kit for testing male fertility comprising a vessel, a base unit, a liquid supply containing liquid and two filters. The first filter is a sample separation filter which forms a hindrance to transmission of spermatozoa. The second filter of the kit is a spermatozoa detection filter comprising a reagent for identifying spermatozoa. Activation of the kit is prevented until a transport medium, such as the liquid, fills a gap allowing spermatozoa to transmit to a detection zone. The kit may be of one-piece construction and utilizes a thin piece of filter material to separate motile from non-motile spermatozoa.
United States Publication No. US 2006/0110821 to David Brickwood entitled "Devices for Motile Sperm Separation" discloses a vessel having an inlet port, an outlet port which is initially closed, a medium into which motile sperm in the sample can migrate into the vessel via the inlet, and an actuator, the operation of which opens the outlet, thereby allowing the medium to flow out of the vessel through the outlet. The medium in the vessel is initially prevented from flowing through the outlet, thus allowing an incubation period which allows motile sperm sufficient time to migrate from the sample into the medium before the medium leaves the vessel. The device preferably comprises a spermatozoa detector in communication with the outlet. In devices for separating sperm, capillary flow takes place through non-fibrous material, such as the space between sheets of closely-juxtaposed material.
United States Publication No. US 2012/0052485 to Lotus Bio (Nymphaea) Ltd. Entitled "Sperm Separation System" discloses a naturally based Sperm Separation System (SSS) for separation of at least a portion of sperm cell populations (SCP) within an original semen sample, such that an enriched SCP sample is obtained, comprising a sperm separation device, comprising a first chamber adapted to contain at least a portion of said original semen sample, said first chamber is characterized by predetermined 3D shape and volume, said first chamber is bounded by a rim such that said original sample is kept below said rim, a second chamber in physical communication with said first chamber and said rim, said second chamber is characterized by predetermined 3D shape and volume, said second chamber is adapted to reside said enriched SCP sample, incubation means, adapted to socket at least one of said cell separation device and to homogeneously thermoregulate the temperature within the same.
U.S. Pat. No. 5,908,380 to Zavos et al. entitled "Compartmentalized Zavos Sperm Swim-Up Column" discloses a device comprising a hollow, vertically supported column, having a closed lower end, and an open upper end. A lowermost or first conical member is arranged at the lowermost end of the column, the lowermost conical member having a lowermost periphery in sealing engagement with the bottom of the column. The first conical member has inclined side walls and a truncated uppermost portion defining a peripheral edge which is open within into the column. A second conical member having a lowermost periphery is also attached to the side walls of the column in a sealing manner therebetween. The second conical member also has tapered walls and a truncated uppermost portion defining a peripheral edge which is open to the inside of the column. The area between the inclined walls of each of the conical members and the inner wall of the column define a periconical area which comprises a compartment for collecting sperm prior to harvesting thereof.
PCT Publication No. WO 2014/043635 to President and Fellows of Harvard College discloses a microfluidic system which includes a microfluidic channel. The microfluidic channel includes a control layer substantially enclosed within a soft polymer, a cell covering element, and a flow channel between the cell-covering element and control layer enclosed within the soft polymer. The control layer is operable to move towards and exert a pressure on the cell-covering element. In the apparatus of WO 2014043635, cells start in one shared cell environment and then get separated. Cell movement can actually be stopped by the exerted pressure forcing a silicone control layer to interfere with the passing of cells in a flow path. The technique described is not based on motility of cells but more directed to flow cytometry where cells in suspension are hydrodynamically driven along a flow path with a focus on fluorescence and staining.
PCT Publication No. WO 2016/063199 to Ecole Polytechnique Federale De Lausanne discloses a microfluidic device for the culture, selection and/or analysis of sample organisms such as nematodes, as well as for other biological entities such as for instance animal embryos. The device features reservoirs, culture chambers and filtering systems allowing for the selection of specific populations/specimens of sample organisms, thus permitting long-term cultures thereof as well as phenotypic/behavioural analyses. This device also requires the use of pressure to operate upon the organisms in a chamber to pass them through a filtering means to a culture chamber.
PCT Publication No. WO 2010/056755 to Craig discloses a robotic microfluidic incubator system which has a thin transparent sidewall and close proximity of the embryo/oocyte/cultured cells to the sidewall allow close approach of a side view microscope with adequate focal length for mid to high power. The arrangement permits microscopic examination of multiple culture wells when arranged in rows (linear or along the circumference of a carousel). Manual or automated side to side movement of the linear well row, or rotation of the carousel, allows rapid inspection of the contents of each well. Automated systems with video capability also allow remote inspection of wells by video connection or Internet connection, and automated video systems can record oft-hours inspections or time lapse development in culture (i.e. embryo cell division progression, or axon growth in neuron cell cultures). As with WO 2014/043635 and WO 2016/063199, above, the device disclosed by Craig requires moving fluid media.
United States Publication No. US 2002/005354 to Spence et al discloses a microfabricated device for sorting cells based on a desired characteristic, for example, reporter-labelled cells can be sorted by the presence or level of reporter on the cells. The device includes a chip having a substrate into which is microfabricated at least one analysis unit. Each analysis unit includes a main channel, having a sample inlet channel, typically at one end, and a detection region along a portion of its length. Adjacent and downstream from the detection region, the main channel has a discrimination region or branch point leading to at least two branch channels. The analysis unit may further include additional inlet channels, detection points, branch points, and branch channels as desired. A stream containing cells is passed through the detection region, such that on average one cell occupies the detection region at a given time. The cells can be sorted into an appropriate branch channel based on the presence or amount of a detectable signal such as an optical signal, with or without stimulation, such as exposure to light in order to promote fluorescence. The apparatus of Spence uses channels with flowing fluids to interact with the microfabricated substrates and relies on reporters of cells and not the cells own movement.
Canadian patent publication No. CA 2752218 to Inguran, LLC., discloses apparatus for sorting stained particles in a fluid stream according to one or more characteristics of the particles. The fluid stream comprises: (A) a fluid delivery system for delivering a fluid stream containing the particles to a first location; (B) an electromagnetic radiation source for delivering a beam of electromagnetic radiation to the first location for exciting particles in the fluid stream to produce fluorescence emissions; (C) an epi-illumination optics system including a focusing lens, where the optics system is operable to direct the beam of electromagnetic radiation through the focusing lens in a forward direction along a beam axis intersecting particles in the fluid stream at said first location so that said particles pass through the beam, resulting in the fluorescence emissions from the particles being directed along said beam axis in a rearward direction; (D) a photodetector for detecting fluorescence emissions from the excited particles; and (E) a processor in communication with the photodetector to classify the particles according to the one or more characteristics based upon the fluorescence emissions of the particles and to sort the particles according to the classification of particles. This system may be applied to staining of sperm cells and subsequent selection under flow cytometry. It also requires moving fluid. In so far as sperm is concerned the system is directed to the characteristics of sperm chromosome content.
European patent No. EP 0958862 to AMS s.r.l. discloses a multiuse reactive-plate for analysis, particularly for direct analysis of whole blood or fluids containing any kind of particles or fragment. The multiuse reactive plate is able to carry out measurements of single analytes. In the system disclosed, fluids are moved by force and involve valves within the flow path.
PCT Publication No. WO 2002/033047 to Nexgenics Bioscience Corp. discloses an embryo support assembly, embryo support system, a method for maintaining the viability of a growing embryo, and a method for shipping a metabolically active embryo. In this apparatus, which uses valves for fluid flow, sperm is only referenced with respect to it being added to the eggs, as is, for fertilisation.
United States Publication No. US2003196714A1 to Gilbert et al discloses a microfluidic system which includes a bubble valve for regulating fluid flow through a microchannel. The bubble valve includes a fluid meniscus interfacing the microchannel interior and an actuator for deflecting the membrane into the microchannel interior to regulate fluid flow. The actuator generates a gas bubble in a liquid in the microchannel when a sufficient pressure is generated on the membrane.
United States Publication No. US 2014/273179 to Inguran, LLC. discloses a device in the form of a microfluidic chip. In particular, various features are incorporated into the microfluidic chip for aligning and orienting sperm in flow channels, as well as for separating selected subpopulations of sperm. Again, this device requires forced flow to move cells. It also is directed to determining sperm chromosome content.
United States Publication No. US 2011/177547A1 to Xia et al discloses a fluidic device which includes an arrangement of channels for introducing a sample containing particles of interest into a processing chamber. The chamber is in fluid communication with collecting channels via low-flow connection channels. Particles in the sample may be observed and diverted from the processing chamber by application of a motive force such as optical trapping into a collection channel. Once in the collection channel, particles can be collected, including by trapping in a porous matrix.
PCT Publication No. WO 2013/129947 to Auckland UniServices Limited discloses a method and apparatus for sorting motile sperm. This described system utilizes the phenomenon of 'edge-trained' motile sperm in which motile sperm with progressive motion that encounter a structure such as a wall will tend to turn and move along the wall in a direction depending on the initial approach angle of the sperm. With the use of this phenomenon the system involves delivering a fluid containing sperm (by way of pumping sperm into channels) into a microvolume at least partially defined by a wall which includes a wall termination or a change in angle away from the microvolume that is noted as being a wall termination, and allowing at least some motile sperm to move along said wall and to exit the microvolume by changing direction away from the microvolume at or near the wall termination, to or towards collection of motile sperm. In another aspect the described system comprises a method for delivering a fluid containing the sperm into a microchannel comprising a wall which includes a wall termination, allowing motile sperm to move in substantially no-flow conditions in the microchannel along the wall and to exit the microchannel by changing direction away from the microchannel at or near the wall termination, to or towards a collection reservoir, and recovery of fluid containing the sorted sperm from the collection reservoir. Specifically, the system as described is based on pumping sperm into channels, stopping, and letting them enter into empty side chambers, then flushing excess sperm away and allowing motile sperm to come back to mid-channels that form the chambers (or conversely; pump sperm to side chambers, flush mid channel and allow motile sperm to come to mid channel and flush again to collect).
PCT Publication No. WO 2003/008931 to Hvichia et al discloses a microscale cell separating apparatus which is able to separate cells on the basis of the size of the cells, interaction of the cells with surfaces of the apparatus, or both. The apparatus comprises a stepped or sloped separation element interposed between an inlet region and an outlet region of a void that can be filled with fluid. The void can be enclosed within a cover, and fluid flow through the void engages cells with the separation element. Only cells which have (or can deform to have) a characteristic dimension smaller than or equal to the distance between a step and the cover or body can pass onto or past a step. Modification of surfaces within the apparatus can also inhibit passage of cells onto or past the step. There is no teaching in this disclosure of its application to sperm or how that may be achieved.
US patent publication No. US 2003/0165812 to Takayama et al discloses a process and apparatus in which motile particles are sorted from non-motile particles in a microfluidic sorting device wherein a stream of sort fluid containing motile and non-motile particles is caused to flow adjacent a media stream in non-turbulent fashion through a sort channel, during which flow motile particles cross the interface between the adjacent flow streams, entering the media stream, and forming a motile particle-depleted sort stream. The sorting devices have use, in particular, for sorting of motile from non-motile sperm. The system described relies on sperm moving in a sheath fluid and traversing across fluids without any apertures or closures.
The following prior art relates to methods involving 'vision tracking' of sperm.
U.S. Pat. No. 8,842,901 to The Regents of The University of California entitles "Compact Automated Semen Analysis Platform Using Lens-Free On-Chip Microscopy" discloses a compact and light-weight lens-free platform to conduct automated semen analysis. The device employs holographic on-chip imaging and does not require any lenses, lasers or other bulky optical components to achieve phase and amplitude imaging of sperm a relatively large field-of-view with an effective numerical aperture of approximately 0.2. A series of digital image frames is obtained of the sample. Digital subtraction of the consecutive lens-free frames, followed by processing of the reconstructed phase images, enables automated quantification of the count, the speed and the dynamic trajectories of motile sperm, while summation of the same frames permits counting of immotile sperm.
U.S. Pat. No. 4,896,967 to Hamilton-Thorn Research entitled "Motility Scanner and Method" discloses an improved motility scanner for characterizing the motion of sperm, bacteria, particles suspended in flowing fluids and the like. The motility scanner includes an improved optical system, a source of illumination for the system and radiation sensing means including electronic image reversal means coupled to the system. A disposable specimen holder allows external loading thereof and its positioning on a heated specimen support. The illumination source functions as a collimator in directing all of the emanating illumination onto the specimen. Both directly transmitted light and light scattered by the specimen are received by an imaging lens and are both focussed thereby onto pixels of a light sensitive device. A movable plate provided with a retarding member is designed to be located at the plane conjugate to the plane of the small source aperture. The directly transmitted light is transmitted through the retarding member and/or the attenuating member. The light scattered by the specimen, however, travels for the most part through the plate in areas not covered by the retarding and/or attenuating members.
U.S. Pat. No. 6,426,213 to Progeny Systems, LLC entitled "Sperm Analysis System" discloses a sperm analysis system which has a sperm sample carrier and "reader" module. The sperm sample carrier includes: a shank defining a chamber with an opening for ingress and egress of a sperm sample; a manually operated pump for aspirating a sample of sperm into the chamber, and a plurality of distinct photon paths intersecting and passing through the chamber. The module includes: a processor responsive to an actuation signal from an operator, a photon source, e.g. a light source, energized by the processor in response to the actuation signal, for sending respective beams of photons through each of the photon paths, a plurality of photosensors, one for each photon path, each for producing a signal indicative of the occurrence and frequency of perturbations in the beam of photons passing through said each's respective photon path and communicating the signal to the processor, and an algorithm run by the processor for processing the plurality of photosensors signals to produce a quantified figure of merit indicative of the motility of sperm within the chamber.
United States Publication No. US 2011/0149287 to M.E.S. Medical Electronic Systems Ltd. entitled "Semen Analysis" discloses a method for measuring the total sperm concentration (TSC) in a sample including: placing the sample in a transparent container between a synchronically pulsed light source and a photodetector; and measuring the optical absorbance of the sample in the range of 800-1000 nm, the TSC of the sample being proportional to the absorbance. Further provided is a sampling device for use in optically analysing a biological fluid, a method for measuring motile sperm concentration (MSC) in a semen sample, a method of determining the average velocity (AV) of sperm cells and a system for analysing semen quality comprising means for measuring TSC, means for measuring MSC; and a video visualization system.
However, the outcome of sperm processing methods comprises a portion of washed, motility-selected sperm, which depending on any given individual clinic's practices requires further processing to be suitable for the needs for IVF, IUI and/or ICSI. This may include further concentration of sperm by centrifugation, counting the sperm concentration and sometimes also determining the motility of the final preparation. It is considered best clinical practice to add a known number of sperm in a known concentration of motile sperm to oocytes. These additional steps add more manual steps to the process, and more sub-samples of the processed sperm to be handled and traced. These concentration counts—and often also the assessments of final motility—are done either manually using hemocytometers or Makler chambers and assessment under microscope, or in laboratories large enough to warrant a separate sperm analysis equipment (computer assisted sperm analysis, CASA), using a CASA instrument such as for example SQA-V Gold by Medical Electronic Systems Global.
Other drawbacks of some known prior art include the following:                Sperm must be removed from the processing vehicle after processing to halt diffusion of components in seminal plasma into buffer solution;        Variability in processing due to multiple human interactions with sample during processing;        No concurrent or linked visual assessment of sperm (concentration, motility, morphology) on current sperm processing platforms;        Separate aliquot of processed sperm for analysis is required        Long processing time exposing sperm unnecessarily long to seminal plasma constituents;        Multiple reagents required;        Multiple consumables required;        Many processing systems subjected to environmental conditions of the laboratory including temperature and ambient atmosphere (either at the bench or in a box incubator).

Method and system for processing a biological sample


A method is for using a skate-grinding device. The method provides an automatic sharpening system (10) that has grip members (14, 16) and a grinding wheel (28). A blade (18) of a skate (20) is inserted into a groove (19). The system is turned on. A motor (44) rotates the grinding wheel (28) and automatically moves the grinding wheel (28) within the groove to grind the blade (18). An electronic unit (86) measures a time the system (10) is used. A user of the system (10) is charged for the time measured by the electronic unit (86).

Ice skates, such as those used in ice-hockey and figure skating, require regular sharpening of the blades to create sharp edges against the ice to prevent slipping when the user skates on the ice. It is very important that the sharpening is performed correctly. Novice users cannot use conventional sharpening devices because they require skills to achieve a satisfactory sharpening of the blade edges. There is a need for an effective an automatic sharpening device that does not require skills to achieve satisfactory sharpening of ice-skate blades.

Automatic sharpening system for ice-skates


The present invention provides moieties that bind to an Ig-like domain, e.g., D4 or D5, of a human receptor tyrosine kinase, e.g., the human Kit RTK or the PDGFR RTK, or the D7 domain of a type V receptor tyrosine kinase wherein the moieties lock the ectodomain of the receptor tyrosine kinase in an inactive state thereby antagonizing the activity of the receptor tyrosine kinase.

Stem cell factor (SCF) is a cytokine that mediates its diverse cellular responses by binding to and activating the receptor tyrosine kinase Kit (also known as SCF-receptor). Kit was initially discovered as an oncogene in a feline retrovirus that captured an activated and truncated form of the surface receptor (Besmer et al. (1986) J Virol 60: 194-203). SCF is encoded by the murine steel (SI) locus while Kit is encoded by the dominant white spotting (W) locus in the mouse (Copeland et al. (1990) Cell 63: 175-183; Huang et al. (1990) Cell 63: 225-233; Flanagan and Leder (1990) Cell 63: 185-194; Tan et al. (1990) Science 247: 209-212; Bernstein et al. (1990) Ciba Found Symp 148: 158-166; discussion 166-172). SCF functions as a non-covalent homodimer and both membrane-anchored and soluble forms of SCF generated by alternative RNA splicing and by proteolytic processing have been described (reviewed in Ashman (1999) Int J Biochem Cell Biol 31:1037-1051). Kit is a member of type-III family of receptor tyrosine kinases (RTK), which also includes PDGF-receptor-α, and β, CSF-1-receptor (also known as M-CSF-receptor or Fms), and the Flt3-receptor (also known as Flk2) (reviewed in Ullrich and Schlessinger (1990) Cell 61: 203-212; Blume-Jensen et al. (2001) Nature 411: 355-365). Kit is composed of a glycosylated extracellular ligand binding domain (ectodomain) that is connected to a cytoplasmic region by means of a single transmembrane (TM) domain (reviewed in Schlessinger (2000) Cell 103: 211-225). The ectodomain of Kit and other members of type-III RTKs all contain five Ig-like domains, in which the second and third membrane distal domains were shown to play a role in ligand recognition (reviewed in Ullrich and Schlessinger (1990) Cell 61: 203-212). Other RTKs whose extracellular ligand binding domains are composed exclusively of multiple Ig-like repeats include members of the VEGF-receptor family (7Ig-like), CCK4-receptor (7Ig-like) and FGF-receptors (3Ig-like). The cytoplasmic region of Kit contains a protein tyrosine kinase (PTK) domain with a large kinase-insert region; another hallmark of type-III RTKs. Binding of SCF to Kit leads to receptor dimerization, intermolecular autophosphorylation and PTK activation. It was proposed that the fourth Ig-like domain of Kit is responsible for Kit dimerization in response to either monovalent or bivalent SCF binding (Lev et al. (1992b) J Biol Chem 267: 15970-15977; Blechman et al. (1995) Cell 80: 103-113). However, other studies have demonstrated that ligand induced dimerization of Kit is driven by bivalent binding of SCF (Philo et al. (1996) J Biol Chem 271: 6895-6902; Lemmon et al. (1997) J Biol Chem 272: 6311-6317).
Characterization of mice mutated at the SCF or Kit loci has shown that SCF and Kit are required for development of hematopoietic cells, melanocytes, germ cells and intestinal pacemaker cells (reviewed in Ashman (1999) Int J Biochem Cell Biol 31:1037-1051). In humans, loss of function mutations in Kit cause the piebald trait that is characterized by de-pigmentation of the ventral chest and abdomen, white fareflock of hair, deafness and constipation (Fleischman et al. (1991) Proc Natl Acad Sci USA 88: 10885-10889). A variety of gain-of-function mutations in Kit were found in different types of human cancers. Activating Kit mutations were found in gastro-intestinal-stromal tumors (GIST), acute myeloid leukemia (AML) and mast cell leukemia (MCL) among other cancers. Mutations were identified in the membrane proximal Ig-like domain (D5) (exon 8 and 9), in the juxtamembrane (JM) domain (exon 11), and in the tyrosine kinase (PTK) domain (exon 17) (see Forbes et al. (2006) COSMIC 2005. BR J. CANCER, 94: 318-22. Somatic mutation database: Catalogue of Somatic Mutations in Cancer http://www.sangerac.uk/genetics/CGP/cosmic/). While there is good evidence that the gain of function mutations in the JM and the PTK domains lead to constitutive activation of Kit, by relieving autoinhibitory constraints (Mol et al., (2004) J Biol. Chem. 279: 31655-31663), the molecular mechanism underlying the gain of function mutations in D5 of the ectodomain is not understood. There is a need to better characterize the structures of RTKs such as Kit and PDGFR, as well as SCF, PDGFα/β, and the bound Kit/SCFcomplex. Such a characterization will lead to the informed identification of regions which may be targeted with drugs, pharmaceuticals, or other biologics.

Inhibitors of receptor tyrosine kinases and methods of use thereof


Apparatus and process for synthesizing a diamond film of high purity at a high rate. Mixture gas of hydrocarbon gas and hydrogen gas is introduced into an arc discharge to produce a gas plasma. This gas plasma is blown against a substrate to deposit diamond. A plasma current power supply is connected with a third electrode disposed above the substrate to place the third electrode at a positive potential. An electrical current is supplied into the gas plasma from the third electrode. Flow of the electrical current through the plasma promotes the decomposition of the hydrogen and hydrocarbon. A diamond film of high purity can be deposited at a high rate. The invention is characterized in that the substrate is placed at a lower potential than the third electrode or the direction of the electrical current intersects the flow of the gas plasma although the electrical current is passed through the plasma. Therefore, it is unlikely that the accelerated electrons reach the substrate, elevating its temperature. Hence, diamond of high quality can be synthesized.

1. Field of the Invention
The present invention relates to an apparatus and process for forming a diamond film and, more particularly, to an apparatus and process for synthesizing a diamond film out of a vapor phase by making use of an arc discharge.
2. Description of the Prior Art
In recent years, various processes have been proposed as methods of synthesizing diamond films out of a vapor phase at low pressures. One process is the hot filament CVD. In particular, a tungsten filament is placed just above a substrate which is heated to 800.degree. to 1000.degree. C. The filament is heated above 2000.degree. C. Hydrogen and a hydrocarbon gas (e.g., CH.sub.4) are blown against the substrate through the filament to grow a diamond film on the substrate.
Another known process is the microwave plasma CVD. In this process, a plasma is produced within a mixture gas of hydrogen gas and hydrocarbon gas by application of microwaves of several hundred watts. A diamond is grown on the substrate placed within the plasma. The substrate is heated to about 700.degree. to 900.degree. C. by the microwaves.
In these two processes for synthesizing diamond, hydrogen atoms act to promote decomposition of CH.sub.4 and to selectively etch synthesized materials other than the diamond such as amorphous carbon. In this way, hydrogen atoms play an important role. However, the hot filament CVD is not practical, because the heated filament often breaks. When the melting point of tungsten is taken into account, the filament cannot be heated above about 2000.degree. C. If the filament is heated above this temperature, the filament wire will break. As a result, the raw material gases cannot be decomposed sufficiently. In the process utilizing a microwave plasma, limitations are imposed on the dimensions of the plasma chamber. Therefore, it is difficult to apply this process to a sample having a large area. In addition, the raw material gases, especially hydrogen, are not decomposed sufficiently.
A further process available uses an ion beam. In this process, a beam of carbon ions is caused to bombard a substrate to grow a diamond film. Unfortunately, the produced diamond contains a large amount of impurities such as amorphous carbon.
A process utilizing an arc discharge has been proposed in U.S. Pat. No. 4,851,254. Specifically, an arc discharge is induced between opposite electrodes. The raw material gases are passed through the arc discharge to create a gas plasma. This gas plasma is narrowed down by a restrictor portion and blown as a plasma jet gas against a substrate. Thus, diamond is deposited on the substrate.
This method of synthesizing diamond by making use of an arc discharge can indeed provide a greatly improved synthesis rate, but the ratio of the hydrocarbon introduced for improving the purity of the diamond film to the hydrogen must be decreased. This lowers the synthesis rate. More specifically, as can be seen from the graph of FIG. 18, as the ratio of the hydrocarbon to hydrogen is decreased, the thermal diffusivity of the synthesized diamond film, or the purity, is improved but the synthesis rate drops.
Another problem with this process using an arc discharge is that the gases are not sufficiently dissociated near the substrate against which the raw material gases are blown. Specifically, the use of the arc discharge greatly improves the gas decomposition rate, but this improvement is accomplished only around the generated arc discharge. The gas decomposition rate is much lower around the substrate against which the plasma jet gas is blown because of recombination and other reactions. In FIG. 19, the spectral intensity of the emission spectrum H.beta. of hydrogen Balmer series representing the amount of the hydrogen radicals, or the decomposed hydrogen, is plotted against the distance from the nozzle, or the plasma ejection port, to the substrate. As can be understood from this graph, the gas decomposition rate drops rapidly away from the plasma ejection port. That is, the decomposition rate is much lower around the substrate.
Bringing the substrate close to the plasma ejection port to synthesize the diamond at positions of high gas decomposition rates may be contemplated. However, this elevates the substrate temperature, rapidly increasing the amount of impurity carbons other than diamond such as graphite. This is because the appropriate temperature of the substrate for the synthesis of a diamond is between 600.degree. C. and 1100.degree. C. and because the optimum temperature for high-purity synthesis is between 800.degree. C. and 1000.degree. C. Therefore, in order to synthesize a diamond at a higher efficiency, it is necessary to enhance the gas decomposition rate around the substrate without elevating the substrate temperature.


A method of making a digital photograph composed of a moving water element and still element by applying a known computer image processing system, and a digital photograph made by this method, involves taking a basic photograph indicating a realistic image of still elements and photographs taken by applying a series of shutter speeds. A series of digital images corresponding to the above-mentioned photographs are indicated on a monitor of the processing system, and an image indicating a realistic image of the moving water element is selected from the series of digital images. A color matching operation is them applied to the selected digital image whereby a final digital image having a still element substantially color matched to that of the basic photograph and a realistic image of a moving water element is indicated on the monitor, and thereafter a realistic digital photograph is printed based upon the final image.

The present invention relates to a method of making a digital photograph including the work of photographing a subject comprised of moving elements and still elements surrounding the same, in particular a subject including flowing water as a moving element, and improving the digital photograph by a computer image processing technique.
In the long practiced art of taking silver film photographs, when photographing a subject comprised of moving elements including water such as a waterfall or rapids and still elements such as the scenery surrounding them, as clear from the numerous photographs (clearly indicating the photographic conditions) described in for example Takao Otsuka, Utsukushiki Nihon no Kawa (Beautiful Rivers of Japan), taking the photograph at the so-called "slow shutter speed" of 1/30 second etc. has been recommended as a condition for photographing such moving elements. The specifications for the recently popularized digital cameras also follow this.
There is however still a doubt as to whether photographs taken by such a traditional photographic technique, leaving aside the aesthetic evaluation, can be said to substantially match with the actual visual perception. The present inventor has long held doubts over photography by this slow shutter speed in view of his experience in taking photographs during studies of the dynamics of high speed rotating objects.
It is known that, in the case of photographing a subject comprising a core formed by a moving water element such as a waterfall or mountain stream and still elements such as trees surrounding the core, if a high shutter speed corresponding to the running speed of the core is applied to take a photograph of such subject, the characteristic feature of the moving element can be captured as a photograph precisely, while the total image of the photograph becomes darker in comparison with the visual recognition by a normal human being. Therefore, the darkness of the photograph of the above-mentioned subject obtained by the high shutter speed cannot generally be accepted, in spite of the effective result indicating a realistic image of the moving element which is obtained by the application of the high shutter speed.
The applicant conceived that, if the above-mentioned problem due to the darkness of photograph can be eliminated under the condition of keeping the realistic image of the moving element, the photograph of the subject comprising the above-mentioned core and the still elements surrounding the core can be accepted in the photographic society. This is the origin of conceiving the present invention.
However, such changes in the darkness of a photograph are also affected by such conditions of the setting diaphragm of lens, and ISO sensitivity of the film used (note: in the case of using a Digital camera, setting value of ISO Sensitivity). Therefore, before arriving at the present invention, the following basic research for confirming the influence of the shutter speed on the darkness of the photograph was carried out in consideration of the influence of the "diaphragm condition", "ISO sensitivity of film used (note: in the case of using a digital camera, setting value of ISO Sensitivity, by four successive experiments.
First Experiment
To achieve the object of the present invention, the present inventor studied what kind of shutter speed was suitable and whether photography by the so-called "slow shutter speed" which had been recommended in the case of photographing rapids, waterfalls, etc. was truly optimal for taking a photograph of a realistic image (first experiment). That is, he took photographs of the scene at the fountain plaza at the Imperial Palace Outer Garden (scene mainly of fountains) using a Nikon F-80 camera and ASA100 film (Kodak E100SW), setting the exposure compensation value to ±0 and the shutter speed divided into nine stages in the range of 1/15 second to 1/4000 second (details shown in Table 1), and using shutter priority AE. He then requested a local photo lab to develop the positive film, read the obtained film by a scanner of a second computer image processing system (owned by the present inventor), stored the digital data in a computer (Macintosh G3), and made a comparative study of the digital images displayed on the monitor. Further, the camera had the function of automatic adjustment of the diaphragm for maintaining suitable exposure. For reference, the diaphragm values corresponding to the shutter speeds are also shown in Table 1.
TABLE 1S: Shutter speed (seconds), V: Diaphragm, *SuitableExposure compensation value limited exceededFIG.SNo.(seconds)V11/15*22(+2)21/30*22(+2)31/6022(+1)4 1/125225 1/250166 1/500117 1/100088 1/20005.69 1/40002.8
TABLE 2EquipmentusedFirst systemSecond systemComputerMacintosh G4Macintosh G3MO driveOlympus Servo MOSame as first640CsystemScannerMicrotec ScanmasterSame as first4systemPrinterEpson 2000 CSame as firstsystemMonitorMitsubishi DiamondSame as firstRD 21Gsystem
Further, for convenience, when printing a digital image displayed on the monitor in the above-mentioned computer image processing system, if using a known color calibration technique, it is possible to easily make a digital photograph of color substantially matching the color condition characterized by brightness, contrast, chroma and color balance of the digital image, so in the following explanation, the "digital image displayed on the monitor corresponding to the digital photograph of FIG. X" is expressed as the "digital image of FIG. X" for simplification of the explanation. The above-mentioned color condition is hereinafter simply expressed as "color."
As clear from FIG. 1 to FIG. 9, when the shutter speed is extremely slow ( 1/30 second or less), the contrast between the moving elements, that is, the fountains, and the still elements (the surrounding trees etc.) falls and the fountains are captured in just the state ofjets of water. As opposed to this, when the shutter speed becomes 1/60 second or more, the contrast between the moving elements and the still elements becomes higher and even the drops of water of the fountains are clearly captured, it is found. Further, it was learned that this change is related nonlinearly with the change of the shutter speed, that is, the change slows at a certain degree of speed or more (in this experiment, 1/500 second). Further, it was confirmed that the slower the shutter speed, the darker the photograph. When viewed visually, not just the state of the jets of water, but the state of the presence of falling drops of water in the jets of water is perceived in reality, so it was confirmed that taking a photograph at the above slow shutter speed is not in line with the object of the present invention. Note that it is self-evident that the same results are obtained even when using a camera not having the function of automatic adjustment of the photographic conditions, if adjusting the diaphragm value and the shutter speed by manual operation based on data clear from this experiment.
Second Experiment
Next, a commercially available digital camera (EOS D-30) was used to take photographs of the scene centered around the fountains in the Imperial Palace Outer Garden in the same way as the above-mentioned photographic experiment at an ISO sensitivity of 100 and shutter speed priority changing the shutter speed to several stages (shutter speeds etc. shown in Table 3). The digital data obtained by this was stored in the computer G4 of the first system shown in Table 2 from a CF card and the digital images (FIG. 10 to FIG. 23) displayed on the monitor.
TABLE 3S: Shutter speed (seconds), V: Diaphragm of lens, *SuitableExposure compensation value limit exceededFIG.SNo.(seconds)V101  *22 111/10*22 121/20*22 131/30*22 141/602215 1/1251916 1/2501617 1/3501318 1/5001119 1/750  9.520 1/1000 821 1/2000  4.522 1/3000  3.523 1/4000  *3.5
In this experiment (second experiment), photographs were taken at a constant of white balance condition exposure compensation value (±0) and a shutter speed set to a shutter speed of 14 stages from 1 second to 1/4000 second. The digital images obtained by this photographic experiment were studied compared with each other by FIG. 10 to FIG. 23. As a result, it was confirmed that the images almost completely matched the results of the above second experiment using a silver film camera. Therefore, a detailed explanation will be omitted. Further, since the camera had an automatic adjustment mechanism for automatically adjusting the diaphragm in accordance with a change in the shutter speed to maintain a suitable exposure, the diaphragm values corresponding to the shutter speeds are also given in Table 3 for reference.
Both when using a silver film camera and when using a digital camera by the above experiment, similar results are obtained, so in consideration of work efficiency, the above-mentioned digital camera was used in the following photographic experiments.
Third Experiment
Further, the third experiment was conducted with the intention of supplementing the above experimental findings.
That is, in this photographic experiment, the ISO sensitivity was fixed to 100, the shutter speed was set to 1/500 second, and the exposure compensation value was adjusted to the five stages of −2, −1, ±0, +1, and +2 to investigate the effects on the photograph quality. In the same way as the above experiments, photographs were taken giving priority to the shutter speed (AE). The changes in the diaphragm value were recorded, whereupon only naturally the diaphragm was set in such condition as 8, 5.6, 4.0, and 3.5. These photographic conditions are shown in Table 4.
TABLE 4S: Shutter speed (1/500 second), E: Exposure compensation valueV: Diaphragm, *Suitable exposure compensation valuelimit exceededFIG.No.EV24−28  25−15.626±04.027+13.5+2*3.5
The results of the comparison of the digital images (monitor display) obtained in this experiment show, as shown in FIG. 24 to FIG. 27, that by moving the exposure compensation value to the minus side, the contrast becomes stronger at the details of the moving elements and the moving elements can be captured realistically as a result, but on the other hand the digital image gradually becomes darker.
Note that with an exposure compensation value of +2, the diaphragm of lens exceeds the minimum limit and the digital image also becomes too white, so this case was omitted from the series of attached drawings.
From the above experimental findings, it was confirmed that in order to photograph in particular a subject comprised of moving elements including flowing water and still elements and realistically capture even details of the moving water element, regardless of whether using a silver film camera or a digital camera, it is necessary to photograph it not by the so-called "slow shutter speed", but by a faster shutter speed and it was confirmed that taking the photograph darker is desirable. Note that it was learned that the suitable shutter speed corresponds to the speed of change over time of the moving water element from the experimental findings of the later explained Embodiments 1 and 2.
According to the results of the above-mentioned experiments, it was confirmed that the dynamic feature of running water, such as a water fall or mountain stream, can be correctly represented as a photograph , if the photograph is taken at a shutter speed corresponding speed of the running water, although the entire photographic image becomes too dark so that the quality of the photograph cannot be accepted. Therefore, it is necessary to solve this problem.
In general, it is well known that the darkness of a photograph (digital image) can be easily corrected by applying a computer image processing technology which is disclosed in software such as Adobe Photoshop 5.5 (Registered Trade Mark), that is, such correction of darkness can be carried out by a simple modification of darkness, or the modification of darkness in relation to contrast, and accordingly by the modification of a tone curve. However, it is quite vague which type of corrections mentioned would be suitable to correct the darkness of the photograph created by applying a high shutter speed, and therefore, the following fourth experiment was carried out to clarify this problem.
Fourth Experiment
The fourth experiment was conducted to throw light on this question. That is, as the original digital image of this experiment and the comparative digital image to be used in the work, the digital images of FIG. 15 and FIG. 22 were selected from the digital images obtained in the above second experiment (for convenience, shown as FIG. 28 and FIG. 29). Further, the darkness of the digital image of FIG. 29, that is, the photograph taken at a shutter speed of 1/3000 second, was compared with that of digital images (FIG. 30 to FIG. 36) obtained by simply adjusting the brightness, adjusting a combination of the brightness and contrast, and adjusting the gradation, including adjusting the contrast for safety's sake, with the objective of correcting them to the brightness of the digital image (FIG. 28). As a result, it was confirmed that while the adjustment of just the brightness or the adjustment of just the contrast is completely unsuitable, adjustment by the latter two can be adopted. Table 5 shows the relationship between the content of the adjustment work and the drawings.
TABLE 5FIG.No.Content of image processing28Original image (same as FIG. 15)29Original image (same as FIG. 22)30Original image 29: Brightness adjustment (+35)31Original image 30: Contrast adjustment (+25)32Original image 32: Brightness adjustment (+30)33Original image 32: Contrast adjustment (+19)34Original image 33: Brightness adjustment (+50)35Original image 34: Contrast adjustment (+22)36OriginalThe brightness of the originalimage 29:digital image (input) and adjustedTone curvebrightness (output) are shown inTable 8 as input values and outputvalues for each of the tone curvesA, B, and C of FIG. 62.
Note that the fact that similar results are obtained by combined adjustment of the brightness and contrast and adjustment of the gradation can be understood from theoretical explanations relating to the parameters for physical evaluation of images, for example, Yoichi Mitake, Dejitaru Karaa Gazo no Kaiseki-Hyoka (Analysis and Evaluation of Digital Color Images) (page 91).
Here, when adopting the method of combined adjustment of the brightness and contrast for adjustment of the brightness of a digital image displayed on a monitor, the problem remains of what extent of adjustment of the brightness to perform first. It was confirmed that this problem is solved by the technique of adjusting the brightness of the original image (FIG. 29) to close to the brightness of FIG. 28 by adjusting the brightness, then adjusting the contrast to make the brightness of the digital image as a whole closely match that of FIG. 28. This fact is proved by the fact that when comparing FIG. 31 and FIG. 33, the brightness and contrast both substantially match.
Further, it was confirmed that it is possible to obtain similar results by adjusting the gradation by a known technique (adjustment of input and output of tone curve dialog box) (FIG. 36). For reference, the data of gradation adjustment is shown in Table 8 with reference to FIG. 62.
Note that FIG. 62 shows a known tone curve added with data display coordinates A, B, and C for the purpose of explaining the detailed content of gradation adjustment.
From this series of experiments, the photographic conditions for preparing a realistic digital photograph starting from a photograph obtained by photographing moving elements including flowing water and still elements and the technique for improving this digital photograph to a realistic digital photograph by computer image processing became clear. This is the crux of the present invention.

Method of making digital photograph and digital photograph made by that method