Nucleic acid able to cause specific cleavage of a bond between two ribonucleotides in an RNA-containing molecule. The RNA-containing molecule has the structure: EQU 5'-XnUHZCUGANGAGYm-3'. wherein each X and Y is independently any nucleotide base; n and m are independently between 5 and 40; H is U, A or C; Z is a hairpin loop, having between 6 and 60 bases, and each U, C, G and A is a uracil, cytosine, guanosine, or adenosine-containing ribonucleotide, respectively, and N is any ribonucleotide. The nucleic acid has the structure: EQU 3'-X'nM.sub.0 Y'm-5'. wherein each X' and Y' are complementary nucleotide bases to each corresponding X and Y, and M.sub.0 is a series of nucleotide bases active to cause the cleavage, and wherein M.sub.0 contains no ribonucleotides.

This invention relates to the cleavage of RNA or DNA by a nucleic acid molecule.
The following is a brief history of the discovery and activity of enzymatic RNA molecules or ribozymes. This history is not meant to be complete but is provided only for understanding of the invention that follows. This summary is not an admission that all of the work described below is prior art to the claimed invention.
Prior to the 1970s it was thought that all genes were direct linear representations of the proteins that they encoded. This simplistic view implied that all genes were like ticker tape messages, with each triplet of DNA "letters" representing one protein "word" in the translation. Protein synthesis occurred by first transcribing a gene from DNA into RNA (letter for letter) and then translating the RNA into protein (three letters at a time). In the mid 1970s it was discovered that some genes were not exact, linear representations of the proteins that they encode. These genes were found to contain interruptions in the coding sequence which were removed from, or "spliced out" of, the RNA before it became translated into protein. These interruptions in the coding sequence were given the name of intervening sequences (or introns) and the process of removing them from the RNA was termed splicing. After the discovery of introns, two questions immediately arose: (i) why are introns present in genes in the first place, and (ii) how do they get removed from the RNA prior to protein synthesis? The first question is still being debated, with no clear answer yet available. The second question, how introns get removed from the RNA, is much better understood after a decade and a half of intensive research on this question. At least three different mechanisms have been discovered for removing introns from RNA. Two of these splicing mechanisms involve the binding of multiple protein factors which then act to correctly cut and join the RNA. A third mechanism involves cutting and joining of the RNA by the intron itself, in what was the first discovery of catalytic RNA molecules.
Cech and colleagues were trying to understand how RNA splicing was accomplished in a single-celled pond organism called Tetrahymena thermophila. They had chosen Tetrahymena thermophila as a matter of convenience, since each individual cell contains over 10,000 copies of one intron-containing gene (the gene for ribosomal RNA). They reasoned that such a large number of intron-containing RNA molecules would require a large amount of (protein) splicing factors to remove the introns quickly. Their goal was to purify these hypothesized splicing factors and to demonstrate that the purified factors could splice the intron-containing RNA in vitro. Cech rapidly succeeded in getting RNA splicing to work in vitro, but something unexpected was occurring. As expected, splicing occurred when the intron-containing RNA was mixed with protein-containing extracts from Tetrahymena, but splicing also occurred when the protein extracts were left out. Cech proved that the intervening sequence RNA was acting as its own splicing factor to snip itself out of the surrounding RNA. They published this startling discovery in 1982. Continuing studies in the early 1980's served to elucidate the complicated structure of the Tetrahymena intron and to decipher the mechanism by which self-splicing occurs. Many research groups helped to demonstrate that the specific folding of the Tetrahymena intron is critical for bringing together the parts of the RNA that will be cut and spliced. Even after splicing is complete, the released intron maintains its catalytic structure. As a consequence, the released intron is capable of carrying out additional cleavage and splicing reactions on itself (to form intron circles). By 1986, Cech was able to show that a shortened form of the Tetrahymena intron could carry out a variety of cutting and joining reactions on other pieces of RNA. The demonstration proved that the Tetrahymena intron can act as a true enzyme: (i) each intron molecule was able to cut many substrate molecules while the intron molecule remained unchanged, and (ii) reactions were specific for RNA molecules that contained a unique sequence (CUCU) which allowed the intron to recognize and bind the RNA. Zaug and Cech coined the term "ribozyme" to describe any ribonucleic acid molecule that has enzyme-like properties. Also in 1986, Cech showed that the RNA substrate sequence recognized by the Tetrahymena ribozyme could be changed by altering a sequence within the ribozyme itself. This property has led to the development of a number of site-specific ribozymes that have been individually designed to cleave at other RNA sequences. The Tetrahymena intron is the most well-studied of what is now recognized as a large class of introns, Group I introns. The overall folded structure, including several sequence elements, is conserved among the Group I introns, as is the general mechanism of splicing. Like the Tetrahymena intron, some members of this class are catalytic, i.e., the intron itself is capable of the self-splicing reaction. Other Group I introns require additional (protein) factors, presumably to help the intron fold into and/or maintain its active structure. While the Tetrahymena intron is relatively large, (413 nucleotides) a shortened form of at least one other catalytic intron (SunY intron of phage T4, 180 nucleotides) may prove advantageous not only because of its smaller size but because it undergoes self-splicing at an even faster rate than the Tetrahymena intron.
Ribonuclease P (RNaseP) is an enzyme comprised of both RNA and protein components which are responsible for converting precursor tRNA molecules into their final form by trimming extra RNA off one of their ends. RNaseP activity has been found in all organisms tested, but the bacterial enzymes have been the most studied. The function of RNaseP has been studied since the mid-1970s by many labs. In the late 1970s, Sidney Altman and his colleagues showed that the RNA component of RNaseP is essential for its processing activity; however, they also showed that the protein component was also required for processing under their experimental conditions. After Cech's discovery of self-splicing by the Tetrahymena intron, the requirement for both protein and RNA components in RNaseP was reexamined. In 1983, Altman and Pace showed that the RNA was the enzymatic component of the RNaseP complex. This demonstrated that an RNA molecule was capable of acting as a true enzyme, processing numerous tRNA molecules without itself undergoing any change. The folded structure of RNaseP RNA has been determined, and while the sequence is not strictly conserved between RNAs from different organisms, this higher order structure is. It is thought that the protein component of the RNaseP complex may serve to stabilize the folded RNA in vivo. At least one RNA position important both to substrate recognition and to determination of the cleavage site has been identified, however little else is known about the active site. Because tRNA sequence recognition is minimal, it is clear that some aspect(s) of the tRNA structure must also be involved in substrate recognition and cleavage activity. The size of RNaseP RNA (>350 nucleotides), and the complexity of the substrate recognition, may limit the potential for the use of an RNaseP-like RNA in therapeutics. However, the size of RNaseP is being trimmed down (a molecule of only 290 nucleotides functions reasonably well). In addition, substrate recognition has been simplified by the recent discovery that RNaseP RNA can cleave small RNAs lacking the natural tRNA secondary structure if an additional RNA (containing a "guide" sequence and a sequence element naturally present at the end of all tRNAs) is present as well.
Symons and colleagues identified two examples of a self-cleaving RNA that differed from other forms of catalytic RNA already reported. Symons was studying the propagation of the avocado sunblotch viroid (ASV), an RNA virus that infects avocado plants. Symons demonstrated that as little as 55 nucleotides of the ASV RNA was capable of folding in such a way as to cut itself into two pieces. It is thought that in vivo self-cleavage of these RNAs is responsible for cutting the RNA into single genome-length pieces during viral propagation. Symons discovered that variations on the minimal catalytic sequence from ASV could be found in a number of other plant pathogenic RNAs as well. Comparison of these sequences revealed a common structural design consisting of three stems and loops connected by central loop containing many conserved (invariant from one RNA to the next) nucleotides. The predicted secondary structure for this catalytic RNA reminded the researchers of the head of a hammer consisting of three double helical domains, stems I, II and III and a catalytic core (FIG. 1a); thus it was named as such. Uhlenbeck was successful in separating the catalytic region of the ribozyme from that of the substrate. Thus, it became possible to assemble a hammerhead ribozyme from 2 (or 3) small synthetic RNAs. A 19-nucleotide catalytic region and a 24-nucleotide substrate, representing division of the hammerhead domain along the axes of stems I and II (FIG. 1b) were sufficient to support specific cleavage. The catalytic domain of numerous hammerhead ribozymes have now been studied by both the Uhlenbeck and Symons groups with regard to defining the nucleotides required for specific assembly and catalytic activity and determining the rates of cleavage under various conditions.
Haseloff and Gerlach showed it was possible to divide the domains of the hammerhead ribozyme in a different manner, division of the hammerhead domain along the axes of stems I and III (see, FIG. 1c). By doing so, they placed most of the required sequences in the strand that didn't get cut (the ribozyme) and only required a UH where H=C, A, U in the strand that did get cut (the substrate). This resulted in a catalytic ribozyme that could cleave a UH-containing RNA sequence embedded within a longer "substrate recognition" sequence. The specific cleavage of a long mRNA, in a predictable manner using several such hammerhead ribozymes, was reported in 1988. A further development was the division of the catalytic hammerhead domain along the axes of stems III and II (FIG. 1d, Jeffries and Symons, Nucl. Acids. Res. 1989, 17:1371.)
Deoxyribonucleotide substitutions in the fragment corresponding to construct 1c have been described by Yang et al., Biochemistry 1990, 29:11156; Perreault et al., Biochemistry 1991, 30:4020; Yang et al., Biochemistry 1992, 31:5005; Usman and Cedergren, Trends in Biochem. Sci. 1992, 17:334; Usman et al, International Publication No. WO 92/07065; Eckstein et al., International Publication No. WO 93/15187; and Rossi et al., U.S. Pat. No. 5,149,796. Jeffries and Symons, supra, indicated that DNA used in construct 1d does not catalyze the cleavage of RNA.
One plant pathogen RNA (from the negative strand of the tobacco ringspot virus) undergoes self-cleavage but cannot be folded into the consensus hammerhead structure described above. Bruening and colleagues have independently identified a 50-nucleotide catalytic domain for this RNA. In 1990, Hampel and Tritz succeeded in dividing the catalytic domain into two parts that could act as substrate and ribozyme in a multiple-turnover, cutting reaction. As with the hammerhead ribozyme, the hairpin catalytic portion contains most of the sequences required for catalytic activity while only a short sequence (GUC in this case) is required in the target. Hampel and Tritz described the folded structure of this RNA as consisting of a single hairpin and coined the term "hairpin" ribozyme (Bruening and colleagues use the term "paper clip" for this ribozyme motif). Continuing experiments indicate an increasing number of similarities between the hairpin and hammerhead ribozymes in respect to both binding of target RNA and mechanism of cleavage. At the same time, the minimal size of the hairpin ribozyme is still 50-60% larger than the minimal hammerhead ribozyme.
Hepatitis Delta Virus (HDV) is a virus whose genome consists of single-stranded RNA. A small region (about 80 nucleotides) in both the genomic RNA, and in the complementary anti-genomic RNA, is sufficient to support self-cleavage. As the most recently discovered ribozyme, HDV's ability to self-cleave has only been studied for a few years, but is interesting because of its connection to a human disease. In 1991, Been and Perrotta proposed a secondary structure for the HDV RNAs that is conserved between the genomic and anti-genomic RNAs and is necessary for catalytic activity. Separation of the HDV RNA into "ribozyme" and "substrate" portions has recently been achieved by Been. Been has also succeeded in reducing the size of the HDV ribozyme to about 60 nucleotides.
The table below lists some of the characteristics of the ribozymes discussed above:
TABLE 1 ______________________________________ Characteristics of Ribozymes ______________________________________ Group I Introns Size: .about.300 to >1000 nucleotides. Requires a U in the target sequence immediately 5' of the cleavage site. Binds 4-6 nucleotides at the 5'-side of the cleavage site. Over 75 known members of this class. Found in Tetrahymena thermophila rRNA, fungal mitochondria, chloroplasts, phage T4, blue-green algae, and others. RNaseP RNA (M1 RNA) Size: .about.290 to 400 nucleotides. RNA portion of a ribonucleoprotein enzyme. Cleaves tRNA precursors to form mature tRNA. Roughly 10 known members of this group all are bacterial in origin. Hammerhead Ribozyme Size: .about.30 to 40 nucleotides. Requires the target sequence UH immediately 5' of the cleavage site. Binds a variable number nucleotides on both sides of the cleavage site. 14 known members of this class. Found in a number of plant pathogens (virusoids) that use RNA as the infectious agent (FIG. 1) Hairpin Ribozyme Size: .about.50 nucleotides. Requires the target sequence GUC immediately 3' of the cleavage site. Binds 4 nucleotides at the 5'-side of the cleavage site and a variable number to the 3'-side of the cleavage site. Only 1 known member of this class. Found in one plant pathogen (satellite RNA of the tobacco ringspot virus) which uses RNA as the infectious agent (FIG. 5). Hepatitis Delta Virus (HDV) Ribozyme Size: .about.60 nucleotides (at present). Cleavage of target RNAs recently demonstrated. Sequence requirements not fully determined. Binding sites and structural requirements not fully determined, although no sequences 5' of the cleavage site are required. Only 1 known member of this class. Found in human HDV (FIG. 6). ______________________________________
The enzymatic nature of a ribozyme is advantageous over other technologies, such as antisense technology (where a nucleic acid molecule simply binds to a nucleic acid target to block its translation) since the effective concentration of ribozyme necessary to effect a therapeutic treatment is lower than that of an antisense oligonucleotide. This advantage reflects the ability of the ribozyme to act enzymatically. Thus, a single ribozyme molecule is able to cleave many molecules of target RNA. In addition, the ribozyme is a highly specific inhibitor, with the specificity of inhibition depending not only on the base pairing mechanism of binding, but also on the mechanism by which the molecule inhibits the expression of the RNA to which it binds. That is, the inhibition is caused by cleavage of the RNA target and so specificity is defined as the ratio of the rate of cleavage of the targeted RNA over the rate of cleavage of non-targeted RNA. This cleavage mechanism is dependent upon factors additional to those involved in base pairing. Thus, it is thought that the specificity of action of a ribozyme is greater than that of antisense oligonucleotide binding the same RNA site.


Methods, compositions, and systems for propagation and fermentation, particularly large scale operations for production of ethanol and dried distiller's grain are provided. Addition of Z. mobilis to propagation and/or fermentation decreases lactic acid bacterial contamination.

The gram negative facultative anaerobic bacterium, Zymomonas mobilis (Z. mobilis) has been considered as a potential alternative to yeast for ethanol production. The use of Z. mobilis has advantages over yeast, including higher specific rate of sugar uptake, lower biomass production, and higher volumetric sugar uptake and ethanol productivity. A commonly known drawback of Z mobilis is that it can tolerate only up to approximately 12 percent weight to volume ethanol (approximately 15.2 percent volume to volume) whereas yeast can tolerate up to approximately 18 percent weight to volume (approximately 23 percent volume to volume).
In a conventional raw starch hydrolysis process, where very high gravity (VHG) fermentation is practiced, a commercially available distiller's yeast is able to produce approximately 15 to 16 percent weight to volume ethanol (approximately 19 to 20.2 percent volume to volume ethanol). In the raw starch hydrolysis process, which has no cooking (liquefaction) step a higher probability exists that the mash may become contaminated by bacteria that produce undesirable end-products.
One substantial source for contaminant bacteria in an ethanol production facility is the feedstock. As much as 10,000 to 1,000,000 bacterial cells per gram enter the facility with the incoming corn. A majority of contaminant bacteria belong to the lactic acid bacterial group since they can adapt and grow well in the ethanol production conditions. Eventually, these contaminant lactic bacteria get established in the production facility if proper measures for control are not taken.
A number of strategies, including the use of low pH (between pH of 4.2 to 4.5) in fermentation, minimal use of antibiotics, and good plant management practices are used to keep the levels of contaminant bacteria to a small quantity. However, natural "non-antibiotic" solutions that eliminate the effects caused by these contaminant bacteria are desirable.
Running antibiotic-free industrial ethanol fermentations is desirable in consideration of global issues surrounding development of antibiotic resistant bacteria.
Wild lactic acid bacteria have a faster growth rate than yeast under ideal conditions. These bacteria, when they are in high numbers in the substrate, can begin to grow rapidly even before yeast is inoculated (added) to the mash. The lactic acid bacteria can easily grow to levels that would produce enough end-products, including lactic and acetic acids, to slow down the efficiency of ethanol production by the yeast, ultimately leading to a loss in ethanol yield.
The present disclosure is directed toward overcoming one or more of the problems discussed above.

Systems, compositions, and methods of fermentation with


A field emission display device is disclosed. Since the size of the cells adjacent to the spacer is set smaller than the size of the other cells, the luminance and aperture rate of the panel can be improved. In addition, the width of the pulse supplied to the cells adjacent to the spacer and the width of the pulse supplied to the cells not adjacent to the spacer are set different, so that the same luminance can be displayed in every cell.

1. Field of the Invention
The present invention relates to a field emission display, and more particularly, to a field emission display device and its driving method that are capable of improving an aperture rate of an overall panel and its luminance.
2. Description of the Background Art
Recently, various flat type display devices are being developed to reduce a weight and a volume of a cathode ray tube (CRT). Such flat type display devices include a liquid crystal display, a field emission display (FED), a plasma display panel, an electro-luminescence, or the like. In order to improve a display quality of the flat type display devices, researches are being actively conducted to heighten a luminance, a contrast and a colorimetric purity.
Among them, the FED is divided into a tip type FED in which electrons are emitted by using a tunnel effect by concentrating a high electric field to an acute emitter, and a flat type FED in which a high electric field is concentrated to a metal with a certain area to emit electrons.
In the tip type FED, electrons are emitted from a conic protrusion portion made of silicon (Si) or molybdenum (Mo) by applying a voltage to a gate electrode to apply an electric field to an electron emission portion.
In the flat type FED, a stacked structure including a metal layer, an insulation layer and a semiconductor layer is formed, wherein electrons are injected into and passes from the metal layer and then emitted outwardly from an electron emission unit.
In the tip type FED, the electron emission amount is determined depending on characteristics of the emitter used for the electron emission. Therefore, every emitter should be fabricated uniform. In this respect, however, it is difficult to fabricate the emitters uniform with the current fabrication process, and in order to fabricate such an emitter, much process time is taken.
In addition, in case of the tip type FED, since the electrons are emitted from the acute emitter, scores of or hundreds of bolt should be applied to a cathode electrode and a gate electrode, causing a problem of much power consumption.
FIG. 1 is a view showing a cell of the flat type FED in accordance with a conventional art.
As shown in FIG. 1, each cell of the flat type FED includes: an upper substrate 101 on which an anode electrode 102 and a fluorescent material 103 are stacked; an electric field emission array 105 formed on a lower substrate 104; and a spacer 109 for supporting the upper substrate 101.
The electric field emission array 105 includes: a scan electrode 108 formed on the lower substrate 104; an insulation layer 107 formed on the scan electrode 108 and a data electrode 106 formed on the insulation layer 107.
The scan electrode 108 supplies current to the insulation layer 107, the insulation layer 107 insulates the scan electrode 108 and the data electrode 106, and the data electrode 106 is used as a fetch electrode for fetching an electron.
The space 109 is installed between the upper substrate 101 and the lower substrate 104. Since a high vacuum state is required between the upper substrate 101 and the lower substrate 104 (to prevent an arcing phenomenon due to an acceleration movement of electrons and a high voltage), the spacer 109 prevents a damage of the panel caused due to a difference between an internal pressure and an external pressure (the difference between an external atmospheric pressure and an internal high vacuum is equivalent to approximately scores of tones).
The flat type field emission display device in accordance with the conventional art constructed as described above will now be explained.
In order to display an image on the display device, first, a negative (−) scan pulse is applied to the scan electrode 108 and a positive (+) data pulse is applied to the data electrode 106. And, a positive (+) anode voltage is applied to the anode electrode 102.
Then, electrons tunnel the insulation layer 107 from the scan electrode 108 to the data electrode 106 and are accelerated toward the anode electrode 102.
The electrons collide with red, green and blue fluorescent materials 103 and excite the fluorescent material 103.
At this time, a visible ray of one of the red, green and blue colors is generated according to the fluorescent material 103.
Compared with the tip type FED, the flat-type FED can be driven at a low voltage since the scan electrode 108 and the data electrode 106 are installed in a facing manner with a certain area.
That is, only a few V to 10V is applied to the scan electrode 108 and the data electrode 106 of the flat type FED, and the scan electrode 108 and the data electrode 106 emitting electrons respectively have a certain area. Thus, compared with the tip-type FED, the scan electrode 108 and the data electrode 106 can be fabricated with a simple fabrication process.
FIG. 2 is a plan view showing a field emission display device in accordance with the conventional art.
As shown in FIG. 2, the FED includes: first and second data connection parts 202a and 202b for receiving a drive voltage from a data driving unit (not shown); a scan connection part 201 for receiving a drive voltage from a scan driving unit (not shown); an anode electrode 102 for receiving a drive voltage from an anode driving unit (not shown); and a connection part 204 for electrically connecting the anode electrode 102 and the upper substrate 101.
The first and second data connection parts 202a and 202b receive the drive voltage from the data driving unit and supply it to the data electrodes, and the scan connection part 201 receives the drive voltage from the scan driving unit and supplies it to the scan electrodes.
The anode electrode 102 is formed within an effective display part 203 of the upper substrate 101, and the anode driving unit applies a few kV high voltage to the anode electrode 102 typically formed as a thin film through the connection part 204.
FIG. 3 is a plan view showing the effective display part.
As shown in FIG. 3, the effective display part 203 includes red cells, green cells and blue cells which are sequentially disposed at regular intervals, and a spacer 109 between cells.
In order to form the spacer 109, a certain space is obtained between the cells. In the region where the space 109 is not formed, the areas between cells are the same each other. Reference numeral 301 denotes an emitter electrode and 302 denotes a fluorescent material 302.
The spacer 109 is divided into a rib type and a cross type. As shown in FIG. 4, there are installed hundreds and thousands of rib type spacers 401 to support a vacuum space between the upper substrate 101 and the lower substrate 104.
Thousands of cross-type spacers 501 as shown in FIG. 5 are installed to support the vacuum space between the upper substrate 101 and the lower substrate 104.
The rib-type and cross-type spacers 401 and 501 are installed between cells (R, G and B). Thus, the cells (R, G and B) are disposed adjacent with a certain space therebetween so that the spacers 501 and 501 can be installed therein.
However, in general, cells are disposed with the certain space (in consideration of formation of the spacer) therebetween, much space loss occurs. That is, since there should be a certain space even between adjacent cells with no spacer formed therebetween, an efficiency of a panel and an aperture rate are reduced.
In addition, since the electron beam is distorted according to the quantity and the position of the spacer 109 within the effective display part 203 (a phenomenon that a proceeding direction is changed as electrons collide with the spacer 109), the brightness of the adjacent cells differs and the angle at which an electron beam spreads is changed to cause a problem that there is a difference in the brightness of a screen.
Moreover, since scores of and hundreds of spacers 109 are formed within the effective display part 203, the aperture rate between the anode electrode 102 and an emitter (not shown) (area occupied by the fluorescent material over an overall area of one cell) is restricted. Therefore, with the disposal of the spacers 109, the aperture rate of the overall panel is degraded, and accordingly, a luminance and efficiency are low.

Field emission display and driving method thereof


An electrostatic clamp or chuck and method uses soft square wave, slew rate limited, A.C. clamping voltages and a balanced, differential drive for clamping flat articles such as semiconductor wafers to pedestals, wafer transfer blades, and the like, with a large ratio of clamping force to clamping voltage, reduced decay of the clamping force and associated nearly constant maximum clamping force, instantaneous elimination of remnant clamping force when the clamping voltage is removed (instant off operation), isolation of the clamped article from the clamping voltage, and substantial elimination of vibration. Preferably the A.C. frequency is 0.1 to 60 Hz. The instant off operation is enhanced by increasing the frequency as the clamping voltage is decreased.

1. Field of the Invention
My present invention relates generally to electrostatic chucks. More particularly, my invention relates to ail electrostatic chuck which uses A.C. clamping voltages and to a method for clamping an article such as a semiconductor wafer to an electrostatic chuck with (1) a large ratio of clamping force to A.C. clamping voltage, (2) suppression of the decay of the clamping forte while the clamping voltage is being applied and (3) substantially instantaneous decay of the remnant clamping force when the clamping voltage is removed; and (4) without substantial vibration of the wafer.
2. Description of the Related Art
Electrostatic clamps or chucks have been used to hold wafers on pedestals, electrodes, etc., for processing in semiconductor wafer fabricating systems, In addition, robots which incorporate electrostatic clamping have been used to load and unload wafers in such systems.
Existing electrostatic clamps or chucks include uni-polar electrode designs which use AC (alternating current) or DC (direct current) excitation. A single electrode, separated from the wafer by a dielectric material, is biased with a grounded AC or DC power supply. The wafer acts as the second electrode and, together with the intervening dielectric and the first electrode, forms a parallel plate capacitor. In a plasma processing reactor, the plasma references the wafer to system ground. Clamping force per volt of applied clamping voltage is high. However, a net charge develops on the wafer and on the chuck, which causes several problems.
First, the clamping force F, cannot be instantaneously turned off. A significant remnant remains, typically for as long as about 30 to 60 seconds after the clamping voltage is removed. As a consequence, once the clamping voltage is removed to permit wafer transfer to or from the pedestal or blade, one must either wait for the remnant clamping force to decay or apply a potentially excessive lifting force to remove the wafer. In short, the choice is between throughput-decreasing delay and potential damage to the wafer.
Also, the net charging of the wafer may damage sensitive integrated circuit devices formed on the wafer.
Alternative electrostatic clamp designs use a pair or multiple pairs of isolated bipolar electrodes which are buried beneath a dielectric layer, thereby isolating the electrodes from the wafer. AC or DC excitation is used. Either AC or DC excitation causes charge separation and/or polarization in the wafer without net charging. However, DC excitation devices suffer from the above-discussed decay of force with time to some low value, as well as the above-discussed turn-off delay when the DC clamping voltage is removed. AC designs experience vibration during the application of the AC clamping voltage. Also, unbalanced AC designs may suffer both clamping force decay and turn-off delay. FIG. 1 depicts the typical decay of the clamping force, F.sub.c, which occurs despite the continued application of the 1000 volt DC clamping voltage that generates F.sub.c. It is not unusual for F.sub.c to decay to about 25% of its original value within 20-30 seconds.
U.S. Pat. No. 4,184,188 illustrates a multiple electrode pair electrostatic wafer clamp and the general principles of electrostatic wafer clamps. The disclosed clamp includes a blade with a multiplicity of parallel interleaved positive and negative paired electrodes arranged on the blade surface. A dielectric layer is disposed over the positive and negative electrodes, forming a multiplicity of capacitors on the blade surface. During electrode energization by a D.C. (direct current) voltage, fringing electrostatic fields are generated between the positive electrodes and the negative electrodes of each capacitor pair. When a wafer is placed over the charged electrodes, the fringing electrostatic forces between the positive and negative electrodes pass through the wafer, creating a clamping force across the wafer and the electrodes by separating charges, so that negative charges in the wafer collect over the positively charged electrodes and positive wafer charges collect over the negating charged electrodes. The clamping force is directly proportional to this electrostatic force acting upon the wafer.
U.S. Pat. No. 4,733,632 discloses a representative prior art wafer transfer apparatus which uses a single electrode pair. The apparatus includes a movable arm, a D.C. electrostatic chuck mechanism having a vertically moving support member attached to the moving arm, and a mechanism for moving the support member. Wafer transport is accomplished by positioning the electrostatic chuck above the wafer to be transferred. The electrostatic chuck or robot has two semi-circular electrodes, which are electrically isolated from one another by an insulating film formed over each electrode and are arranged to fit over the periphery of the top surface of the wafer to be clamped. (The periphery of wafers generally is not processed, and therefore provides a clamping surface.)

Isolated electrostatic chuck and excitation method


The positioning device can position a spherical object. This object is placed and held by gravity on three points of support (P, P′, P″) of the positioning device. The center of gravity of the object in any position is within the triangle formed by the three points of support. The positioning device includes means for driving the object in rotation. The drive means includes a drive member including wheel whose contact with the external surface of the object forms one of the support points (P) of the positioning device. This drive member can impose a rotation on the object, via the wheel, in all directions. The other two points of support (P′, P″) are made using two ball and socket joints, wherein the ball is held on an air cushion in a housing of the ball and socket joint.

The use of positioning devices, particularly for systems measuring the radiation diagram of a transmitting antenna, is known. This positioning device can orient, for example, the transmitting antenna, in all directions relative to a fixed receiving antenna, with well-known features of a receiver device. This receiver device, which is connected to a data processing station, can pick up the electromagnetic field generated by the transmitting antenna some distance away, in every position or orientation of the antenna. In this way, the processing station can determine the radiation diagram of the transmitting antenna so as to determine the features of the antenna.
The transmitting antenna to be measured can be placed in a spherical element, so that the spherical element can be held by gravity, for example, on three support points of a positioning device. This positioning device of the prior art includes three drive wheels. The contact of each wheel on the external surface of the spherical element constitutes one of the three support points. These drive wheels can be small tyres, which are each set in motion by drive wheels, such as compressed air turbines or motors. The driving of the wheels causes the spherical element, in contact with the wheels, to rotate randomly in three directions.
With the use of three drive wheels, the positioning device of the prior art has some drawbacks. Indeed, when the three drive wheels rotate, the movement, which they communicate to the spherical element, involves a movement tangential to the support points, i.e. to the three contact points. This movement is the result of three wheel friction forces. In other words, the wheels skid most of the time on the external surface of the moving spherical element. The effect of this is to animate the spherical element in a chaotic manner.
The various frictions act like rubbers on the external surface of the spherical element, dirtying the external surface of the spherical element. Since, in some cases, the tangential movement of the sphere is exactly orthogonal to the orientation of one of the wheels, this therefore means that the spherical element jumps around its origin. This momentary deviation of the centre of the sphere, leads to an error in the measured position or orientation of the transmitting antenna, which constitutes a drawback. Moreover, during the various jumps, the spherical element, which may be off-balance, has a marked tendency to fall back along its mass centre. This results in a lack of homogeneity in the positions travelled.
U.S. Pat. No. 3,441,936 describes a device for positioning a spherical element in which a transmitting antenna can be placed. This spherical element is held, by gravity, on three drive wheels of the positioning device that are capable of orienting the transmitting antenna in any direction. When one drive wheel is being driven, in order to move the spherical element, the spherical element skids or jumps onto the other two drive wheels. This means that proper positioning of the transmitting antenna cannot be guaranteed, which is a drawback.

Device for positioning an object in all directions

The Swatch Group Research and Development Ltd


Structural support members are surrounded with a cover that insulates the outer envelope of the support member. The cover may be made of an elastomeric material sufficiently flexible to permit the cover to be physically installed about the structural member. The structural member and cover may also be provided with interlocking means that will retain the cover on the support member once it is installed. Disclosed is a skylight framework formed of a plurality of spaced rafters joined by a plurality of crossbars wherein the rafters and crossbars each include an elongated central support member having a cover member that surrounds the outer envelope of the elongated support member except for a central fastener opening.

This invention relates to skylight frameworks, and particularly to a framework of rafters and crossbars that insulates and cushions the framework with respect to the glazing panels that is supports and the structural frame upon which it is mounted.
Frameworks for the glazing panels that form a skylight are typically built up from extruded aluminum rafters and crossbars that are joined together in a variety of ways using threaded fasteners, clips, and the like. The glazing panels are typically pre-assembled in a factory with one or more glazing sheets in a panel and some supporting structure as part of the panel. This allows ease of installation directly to the framework. The framework may provide the necessary structural support for all of the weight and loads on the skylight. However, more typically the framework is itself mounted to a structural frame which may be built up from sections of steel for added strength and durability.
The aluminum framework is subjected to many temperature changes between the exterior and the environment that is enclosed by the skylight. The temperature changes can result in significant condensation of moisture on the surface of the aluminum elements. Heretofore, it has been typical to accommodate condensation by the use of gutters that are extruded into the outer surfaces of the aluminum elements to collect moisture and carry it away. An example of such gutters is shown in U.S. Pat. No. 5,092,087 issued Mar. 3, 1992, and assigned to the assignee of this application.


A video processing system includes a plurality of identical video switch modules (A). Each of the video switch modules has matching inputs and outputs such that received signals can be passed through unchanged, enabling the modules to be connected in a daisy chain or series relationship. Each of the modules further includes inputs and outputs for receiving video signals from and sending video signals to procedure rooms (B). Preferably, each of the procedure rooms includes medical diagnostic apparatus (10-18) which produce one or more unprocessed video signals, a first video monitor (22) for displaying first processed video signals, a second monitor (24) for displaying second processed video signals, and a VCR (20) for recording and playing back second processed video signals. The daisy chain of video switch modules is interconnected at one end with a shared digital image processing system (C). Unprocessed video signals from a selected one of the procedure rooms are connected by its corresponding video switch module and the other daisy chained modules to the digital image processing system. The digital image processing system processes the received video signals and images from memory (34) to provide at least first and second processed video signals that are sent back through the daisy chained switch modules to the selected module.

The present invention relates to the processing and handling of diagnostic video image data. The present invention finds particular application in conjunction with vascular diagnostic imaging systems and will be described with particular reference thereto. However, it is to be appreciated, that the invention may also find application in conjunction with the handling of other diagnostic image data.
In a vascular diagnostic system, an x-ray source is commonly disposed on one side of the patient and an image intensifier is disposed on the other side of the patient. The image intensifier converts the x-rays which are transmitted through the patient and strike the image intensifier into a corresponding optical image. That is, an optical or light image is generated in which the intensity of light at each point of the image varies in accordance with the intensity of radiation which passed through the patient and struck the image intensifier. A beam splitter is mounted to the image intensifier for selectively directing the optical image to one or more cameras. The cameras typically include video or electronic cameras and film cameras for generating single frame images and motion picture or video images.
The video images from the electronic camera are typically acquired and processed by a digital image processing system. Each diagnostic suite includes appropriate video monitors for displaying the digitally processed video images and a VCR for recording video electronic images. The VCR playback images may also be displayed in the diagnostic suite or on a control room reference image monitor. In addition to the current or live data from the video camera, the diagnostician commonly wishes to view one or more reference images. The reference image may be an image taken several minutes earlier, an image from an earlier examination, or the like. One common application of the reference image is in conjunction with radiation opaque dyes. After a reference image is recorded, the patient is injected with the radiation opaque dye. Both the live images and the recorded reference images are then displayed on the live and reference image monitors, respectively. The two images are to be compared for diagnostic purposes.
One of the problems with such systems is that the various electronic image signals do not have the same bandwidth. Typically, the live image is a high resolution image of 1024.times.1024 pixels or better, displayed at 30 or higher fps, in either progressive or interlace format. An S-type VCR records or plays back images with a much lower resolution, e.g. about 512.times.512 pixels per image at 30 fps interlaced. These lower resolution images are conveyed with a lower bandwidth signal. Typically, the VCR playback images are displayed on the reference image monitor which is usually dual rate type, capable of displaying both high resolution and S-type VCR images.
Typically, the output of the electronic camera is digitized and conveyed to a digital processing board which generates a low bandwidth downscan signal for recording on a VCR and a high bandwidth live signal for display on a high resolution live image monitor. Further, the digital processor board converts reference images stored in a video memory into high bandwidth reference signals for display on the reference image monitor.
Typically, a system may include more than one diagnostic imaging room as well as a central control console. The central control console and the digital processor boards are relatively expensive. Rather than provide each room with its own digital processor board, a significant cost savings can be achieved by sharing the digital processor board. This requires a relatively complex switching system for switching the camera outputs and the VCR outputs from each room to the digital processor board and the outputs from the digital processor board back to the appropriate monitors and VCRs in each room and the control console.
In a vascular diagnostic system with a large number of rooms, either a larger number of digital processor boards are required or an increasingly complex switching system. One of the problems with the prior art is that when a hospital adds additional rooms, a new switching system is required which can switch the digital processing board among a larger number of rooms. Analogously, manufacturers must inventory a large number of different size switching systems in order to accommodate the varying number of rooms which hospitals might order.
The present invention provides a new and improved daisy chain video switch system which overcome the above-referenced problems and others.


An interaction system, including an antibody or, antibody fragment having functional capability, which comprises a surfactant-stabilized microheterogeneous dispersion of aqueous phase in a water-immiscible medium, said aqueous phase containing an amount of said antibody or said fragment in a functional state sufficient to effect the interaction; and methods for making and for using said system.

While living systems are often viewed as the reaction of biochemicals in water, considerable research has been devoted to the study of the structure and function of biochemicals in mixed water-organic solvent systems. In 1969, Kaminsky and Davidson (1) (see bibliography concluding this section for citations to referenced publications) examined the effects of organic solvents on the low molecular weight protein cytochrome c (12K daltons). A similar approach was used by A.A. Klyosov et al. (2) and Bettelheim and Lukton (3) to measure the enzymatic activity of small molecular weight proteins.
Gitler and Montal (4) radically departed from these early studies in their stabilization of cytochrome c in a solvent system composed primarily (90%) of a water-immiscible solvent, decane, by complexation with a biological detergent, i.e., phospholipids. Marinek et al. (5) explored the stabilization and subsequent enzymatic activity of another low molecular weight protein, chymotrypsin (22K daltons), in iso-octane. The anionic detergent used by Marinek et al. was a non-biological mimic of phospholipids, namely sodium bis-(2-ethylhexyl)-sulfosuccinate ("AOT"). Similarly, Luisi et al. (6) found that chymotrypsin, and other low molecular weight enzymes, trypsin and pepsin (23K and 35K daltons, respectively), retained their enzymatic activity in the water-immiscible solvent cyclohexane when complexed with the cationic detergent methyltrioctylammonium chloride. Luisi et al. suggested that those enzymes retained their basic natural structure within a small aqueous core surrounded by a sphere of detergent and organic solvent. Such structures were termed "reverse micelles", forming within water-immiscible solvents when detergent content is above the critical micelle concentration ("CRC") and when the molar ratio of water to detergent ("W.sub.o ") is in the range from 5:1 to 50:1, inclusive. The bulk of these solutions (80-90%) is composed of a water-immiscible organic solvent.
Physical evidence tending to confirm the existence of reverse micelles has been gained from quasi-elastic light scattering, Huang and Kim (7), and small-angle neutron scattering measurements, M. Kotlarchyk et al. (8). Structural and mechanistic information concerning proteins within reverse micelles has been obtained through NMR (DeMarco et al. (9)), ultraviolet, circular dichroism and fluorescence (P.L. Luisi, 1979 (6)) spectroscopies, as well as enzymatic kinetic analyses.
A variety of low molecular weight proteins and enzymes have been stabilized in a variety of solvents and with a variety of surfactant and co-surfactant systems (P.L. Luisi, 1985 (10)). Although a diverse set of conditions was used in these experiments, some conclusions can be drawn. First, these studies were largely performed with relatively low molecular weight proteins (e.g., cytochrome c (12K daltons), ribonuclease (13K daltons), lysozyme (14K daltons), chymotrypsin (23K daltons), trypsin (35K daltons), alcohol dehydrogenase (37K daltons), rhodopsin (38K daltons), and lipase (45K daltons)), when compared to the molecular weight of the immunoglobulin molecules (150K daltons). Second, the majority of these proteins displays maximal catalytic activity at a water-to-detergent ratio (W.sub.o) of about 10-15. (We note that lipase of 100K daltons has been reported, and that this enzyme has been said to have an activity which increases up to W.sub.o of about 30 and then levels off with (Luisi et al. (12)); however, the molecular weight of this enzyme is significantly less than that of an antibody as contemplated by the present invention, and further the enzyme is lipophilic rather than hydrophilic, and thus its behavior should not be viewed as indicative of antibody behavior. And finally, as suggested by P.L. Luisi et al. (6), "Another interesting property of the phase transfer process . . . is its selectivity towards certain proteins" (p. 751). Factors such as protein size and charge, solution pH, surfactant composition and concentration as well as ionic strength, determine if a particular type of protein can be successfully stabilized within reverse micelles. Particular combinations of these factors are specific to different types of proteins, and this undercuts the predictability of whether one protein type can be stabilized from the finding that a different protein type can. Accordingly, it would not have logically followed that the teachings of the aforementioned workers could be extrapolated beyond the specific conditions and materials utilized by them.
It is also important to note that while stabilization of enzymes within water/water-immiscible biphasic systems was explored to catalyze reactions with water-insoluble substrates (Larsson (11); P.L. Luisi et al. (12)), or for mimicking physiological conditions (Han and Rhee (13)), there are other applications of reverse micelles such as both liquid and HPLC chromatography (M.A. Herandez, 1986 (14)), with supercritical fluids (R.W. Gale et al., 1987 (15)) and in protein extraction (Goklen and Hatton, 1985 (16)).
However, reports of antibody stabilization in these water-immiscible solvents (both organic liquids and supercritical fluids) are extremely limited.
Karr et al. (17) examined a covalent modification of IgG molecules with polyethylene glycol (PEG) as a method of improving the solubility of antibodies in a dextran phase of a biphasic solution. While a dextran phase is far more water-miscible than iso-octane or benzene, the major drawback of this approach is the extra time-consuming steps required for forming the PEG-antibody covalent complex and the possibility that chemical modification of a protein with PEG could also covalently modify amino acid residues in the antigen binding site, thereby reducing the antibody's antigen-binding capacity. In addition, there is no guarantee that this technique works with water-immiscible solvents.
Eremin et al. (18) examined the effects of antibodies directed against horseradish peroxidase in 50 mM aqueous AOT solutions in heptane. The thrust of this work was to study the loss of peroxidase activity observed due to mixing of enzyme and polyclonal anti-peroxidase antibodies. But there is no indication that the antibody was of a monoclonal nature or had a molecular weight which would have been conducive to solubilization in the reverse micelles at the conditions utilized, that the enzyme was inactivated by the antibody, or even that the system of Eremin et al. contained an effective amount of antibody in a functional state. Significantly, no attempt was made to test the antibody's antigen-binding capability in an ELISA analysis after solubilization in iso-octane. Since the presence of large antibodies in the polyclonal mixture, such as those of the IgM nature (970K daltons), could well have caused the applied conditions to be unfavorable for solubilization of antibodies or horseradish peroxidase, this article would not have disclosed use of an effective amount of functional antibodies in an aqueous/non-aqueous biphasic system to the skilled worker.
The development of information concerning conditions for stabilizing an effective amount of antibody or antibody fragment in a heterogeneous system comprising a dispersed aqueous phase, with retention of an effective amount of antibody or antibody fragment in a functional state, would be a significant step forward in the art.
(1) L.S. Kaminsky and A.J. Davidson, FEBS Letters 3, 338 (1969). PA1 (2) A.A. Klyosov, N. Van Viet, and I.V. Berezin, Eur. J. Biochem. 59, 3 (1975). PA1 (3) F.A. Bettelheim and A. Lukton, Nature 198, 357 (1963). PA1 (4) (a) C. Gitler and M. Montal, Biochem and Biophys Res. Comm. 47, 1486 (1972); (b) FEBS Ietters 28, 329 (1972). PA1 (5) K. Martinek, A.V. Levashoft, N.L. Klyachko, and I.V. Berezin, Doklad Ada Nauk SSSR (Engl. edit), 236, 951 (1978). PA1 (6) P.L. Luisi, F.J. Bonner, A. Pellegrini, P. Wiget, and R. Wolf, Helv. Chim. Acta 62, 740 (1979). PA1 (7) J.S. Huang and M.W. Kim, Phys. Rev. Lett. 47, 1462 (1981). PA1 (8) M. Kotlarchyk, S.-H Chen, J.S. Huang, and M.W. Kim, Phys. Rev. A 29, 2054 (1984). PA1 (9) A. De Marco, E. Menegatti, and P.L. Luisi, J. Biochem and Biphys Meth. 12, 325 (1986). PA1 (10) P.L. Luisi, Angew Chem Int Ed Engl. 24, 439 (1985). PA1 (11) K. Larsson, P. Adlecreutz, and B. Mattiasson, "Biocatalysis in Organic Media," C. Laane, J. Tramper, and M.D. Lilly, Eds., Elsevier Science Publishers B.V., Amsterdam, p. 355 (1986). PA1 (12) P.L. Luisi, P. Luthi, I. Tomka, J. Prenosil, and A. Pande, Annals N.Y. Acad. of Sci. 434, 549 (1984). PA1 (13) D. Han and J.S. Rhee, Biotech and Bioeng 28, 1250 (1986). PA1 (14) M.A. Hernandez-Torres, J.S. Landy, and J.G. Dorsey, Anal. Chem. 58, 744 (1986). PA1 (15) R.W. Gale, J.L. Fulton, and R.D. Smith, Anal Cheml 59, 1977 (1987). PA1 (16) K.E. Goklen and T. Alan Hatton, Biotech Progress 1, 69, (1985). PA1 (17) L.J. Karr, S.G. Shafer, J.M. Harris, J.M. Van Alstine, and R.S. Snyder, J. Chromatography 354, 269 (1986). PA1 (18) A.N. Eryomin, M.I. Savenkova, and D.I. Metelitsa, Bioorg. Khim 12, 606 (1986).

Interaction system comprising a surfactant-stabilized aqueous phase containing an antibody fragment


A vegetation cutter for cutting vegetation circumferent an obstacle. The vegetation cutter comprises a frame comprising a dock for receiving the obstacle; at least one cutter that is moveable in a cutting trajectory whereby vegetation circumferent a docked obstacle is cut; and a driver for moving the at least one cutter in the cutting trajectory. A method for cutting vegetation circumferent an obstacle.

Roadsides in many countries must be lined with delineators (reflectors) at set intervals and traffic management signs (e.g. speed zone signposts) to guide drivers. These delineators and signs must be visible from a distance and hence cannot be covered by vegetation. Regular cutting of vegetation on roadside verges is therefore carried out to maintain appropriate sightlines, as well as to manage bushfire risk and to ensure that roadside vegetation does not impede water runoff after heavy storms.
Currently, vegetation on roadside verges is typically cut using a conventional slasher mounted on the back of a tractor. However, this arrangement does not allow for the cutting of vegetation close to delineator posts etc., with attempts to do so often resulting in damage or destruction of the posts. As such, it is often necessary for people to subsequently walk the route with whipper snippers and/or chemical vegetation spray management units to remove the remaining vegetation.
Mowers for cutting vegetation around posts and the like have been disclosed in the prior art. For example U.S. Pat. No. 3,241,302 discloses such a mower. However, such mowers are actuated by the post itself, as the mower moves past it, which can result in the post being damaged. It would be advantageous to provide alternative vegetation cutters for cutting vegetation around an obstacle.
It is to be understood that, if any prior art information is referred to herein, such reference does not constitute an admission that the information forms part of the common general knowledge in the art, in Australia or any other country.

Vegetation cutter with a dock for receiving obstacles


A method of manufacturing a nonvolatile memory device including forming on a lower insulating layer a first sacrificial pattern having first openings extending in a first direction, forming a second sacrificial pattern having second openings extending in a second direction on the lower insulating layer and the first sacrificial pattern wherein the second openings intersect the first openings, etching the lower insulating layer using the first and second sacrificial patterns to form a lower insulating pattern having contact holes defined by a region where the first and second openings intersect each other, forming a bottom electrode in the contact holes, and forming a variable resistance pattern on the lower insulating pattern so that a portion of the variable resistance pattern connects to a top surface of the bottom electrode.

1. Field of the Invention
The present inventive concept relates to a method of fabricating a nonvolatile memory device, and more particularly, to a method of fabricating a nonvolatile memory device using a resistive element.
2. Description of the Related Art
Examples of nonvolatile memory devices which use a resistance material may include a phase change random access memory (PRAM) device, a resistive random access memory (RRAM) device, and a magnetic random access memory (MRAM) device. Such nonvolatile memory devices which use a resistance material may store data by using variations in the state of a phase change material, for example, a chalcogenide alloy, variations in the resistance of a variable resistor, or variations in the resistance of a magnetic tunnel junction (MTJ) thin film with respect to the magnetization of a ferromagnetic material. Whereas Dynamic random access memory (DRAM) devices or flash memory devices may store data using charge.
For example, PRAM devices define a crystalline phase of a phase change material as a set state or a logic value of "0" and define the amorphous phase of a phase change material as a reset state or a logic value of "1." This is due to the fact that a phase change material has a lower resistance in the crystalline phase and has a higher resistance in the amorphous state. In PRAM devices, a write pulse, for example, a set pulse and/or a reset pulse, may be provided to a phase change material, and data may be written to the phase change material with joule heat generated by the write pulse. For example, PRAM devices may apply a write pulse, for example, a set pulse and/or a reset pulse, to phase change materials and write data to the phase change materials using joule heat generated by the write pulse. PRAM devices may write a logic value of "1" to phase change materials by heating the phase change materials to their melting point or higher using a reset pulse and quickly cooling the phase change materials so that the phase change materials become amorphous. PRAM devices may write a logic value of "0" to phase change materials by heating the phase change materials to a temperature between the crystallization temperature and the melting point using a set pulse, and, then, maintaining the temperature of the phase change materials for a given amount of time so that the phase change materials may become crystalline.

Method of fabricating nonvolatile memory device


An airflow guard for a microphone includes a shell having an airflow diverter and a flow separation edge. The airflow diverter partially surrounds the microphone to redirect a flow of air away from the microphone. The flow of air separates from one of the airflow diverter or the flow separation edge. An acoustic reception system for a mobile vehicle and a hands-free communication device are also disclosed.

Automobile manufacturers and designers have focused on airflow smoothening and efficient sound-insulating methods for reducing noise in the vehicle cabin. Noise sources such as the wind, turbulence, and pressure fluctuations can excite the vehicle body and transmit noise into the car cabin. Other inherent noises of the automotive environment include tire and engine noise, as well as voices of other passengers. Fans and blowers of the heating, ventilation, and air conditioning systems generate noise and also generate local pressure variations in the forced air stream.
Besides being interested in finding ways to reduce the generation of turbulence, automotive manufacturers recognize the need to reduce the influence of the air pressure fluctuations inside a vehicle cabin upon various audio components such as a microphone of an in-vehicle cellular phone or a voice-recognition system.
Some of the newer automobile microphone systems use electronic processing, multiple microphones, or both to reduce the influence of the pressure fluctuations. These microphones can be located on rear-view mirrors, headliners, or steering columns.
In one example, an in-vehicle microphone system located in an overhead console picks up the driver's voice and uses algorithms in its electronic processing to cancel an "echo effect" and reduce background noise. This electronic processing helps improve the transmission quality of the driver's speech.
In another example, a self-contained digital-signal-processing (DSP) microphone system uses a digital microphone array and software algorithms to help reduce voice recognition and audio intelligibility issues common in high noise, automotive environments.
Microphone systems for vehicles would be improved if the influence of airflow within the cabin was reduced and the system did not require multiple microphones or signal-processing software to electronically reduce the influence of pressure fluctuations produced by in-vehicle airflow, thereby increasing the signal-to-noise ratio and improving the fidelity of the microphonic pickups to improve clarity of speech. Therefore, an improved in-vehicle microphone system provides clearer voice recognition, increases speaker intelligibility, enhances other noise reduction techniques, and reduces packaging complexity, circuitry and costs, while minimizing the influence of airflow around the vehicle cabin.


A system providing transitional bracket and transfer jig modules that have been custom designed to address a rotated or partially erupted tooth in advance of applying a computer aided bracketing system to the tooth. The transfer jigs and brackets are used until the problem tooth is moved into a position that will allow positioning of the “ideal” bracket. Once the ideal brackets are position, the treatment plan can proceed as it would with existing systems to reach the final ideal position. The transitional modules are integrated into the customized treatment plan such that the orthodontist just follows the digitized plan, including correction of misaligned teeth. The computer system that assists with defining the treatment plan takes into account the starting position of the teeth and can adjust the plan, as well as use of transitional brackets, to accommodate many more patients that have one or more problem teeth needing correction.

Existing digital orthodontic systems utilize digitized images of the teeth to generate a customized treatment system of brackets, transfer jigs and wires. This system assumes that all of the custom designed brackets and transfer jigs can be placed on the facial or labial surface of the tooth. Unfortunately, for some patients a tooth may be rotated or otherwise malpositioned such that the facial surface is not facing outward or the tooth has not fully erupted through the gums. In these situations, the orthodontist must first recognize the problem condition and then use manual techniques to de-rotate or otherwise move the tooth so that the "customized" system can then be placed. Given the cost of such customized systems, it then becomes less desirable to use a customized orthodontic set because the orthodontist must use traditional techniques in addition to the customized versions.
The devices, systems, and methods disclosed herein overcome one or more of the deficiencies of the prior art.

Sequential modules for computer aided bracketing systems and associated methods and devices


A terminal position assurance system is provided for an electrical connector which includes a dielectric housing having a terminal-receiving passage into which a terminal is inserted in an insertion direction to a fully inserted position. A latch surface is formed in the housing adjacent the passage and facing in the insertion direction. A bendable tab is provided on the terminal and is adapted to be bent against the latch surface. The bendable tab is structured to be bent about an axis generally parallel to the insertion direction to move an edge portion of the tab to a position over the latch surface when the terminal is in its fully inserted position.

Terminal position assurance systems heretofore employed in electrical connectors often require the use of a separate component to provide the terminal positioning assurance. In other words, after terminals are inserted into terminal-receiving passages in a connector housing, a separate component is inserted into an opening or openings in the housing which communicates with the terminal-receiving passages. In essence, this separate component provides a secondary lock on the terminals as well as some indication that all of the terminals are in their proper positions.
Problems are encountered with the use of separate terminal position assurance components in that they are more susceptible to leakage or arc tracking. In addition, because of the additional separate components, a user of such systems must maintain an inventory of not only the connector housings and the terminals, but also the separate terminal position assurance or locking components. The potential of leakage or arc tracking is created because the separate components require an opening in the housing for access to a portion of a terminal.
This invention is directed to solving the above problems and providing an improved terminal position assurance system which does not require separate locking components and, in fact, uses an integral portion of each terminal as a secondary locking feature which provides the terminal position assurance system. The integral portion herein is provided by a bendable tab.
Bendable tabs have been used heretofore for locking terminals in respective terminal-receiving passages within connector housings. However, such prior art bendable tabs have not been capable of providing a terminal positioning assurance system.
More particularly, the bendable tabs of the prior art are generally planar and are bent about axes or fold lines which are generally perpendicular to the insertion direction of the terminals. Such bendable tabs are bent to overlie locking surfaces of the connector housing adjacent the terminal-receiving passages. In other words, the tabs are bent about axes or fold lines generally parallel to such surfaces. Consequently, a particular terminal might not be fully inserted into its respective terminal-receiving passage, but the bendable tab still is capable of being bent to overlie the locking surface, notwithstanding the fact that the terminal is not properly positioned in the housing.
The present invention utilizes a simple bendable tab which not only can be considered a terminal locking feature but also provides a terminal-position assurance means for the terminal.

Terminal position assurance system for electrical connectors


A security feature in a handheld electronic wireless communication device, has two switch buttons A and B that are positioned on the bezel of the device that are for increasing/decreasing functions of the device. The device has a LED light used for flash light in a camera mode of the device. A device-logic, in a non-active state of the device, detects activations of the two switches in the non-active state of the device and uses the switch activations to control the LED light; thereby the logic enables quick response use of the LED light for security purposes.

Wireless mobile communication devices, also known as smart phones, have become very common and have been acquired and are carried in their personal possession by the masses due to their low cost, convenience, and functionality. These wireless devices come equipped with features such as speakerphone, camera and GPS location.
Hence it is the objective of the embodiments herein to add more features in the mobile devices making them even more useful and convenient to the users of these devices.

Apparatus and method for a one touch light security feature in a mobile wireless communication device


To provide an automatic report creation system which can automatically create reports for various different uses. This automatic report creation system included a voice recognition unit 3, a clock unit 7 and a term dictionary 9, wherein the voice recognition unit 3 recognizes the input of a pre-stored start term, the clock unit 7 calculates the start time on the basis of the time when the voice recognition unit 3 recognized input of the start term, the voice recognition unit 3 recognizes that a pre-stored end term has been inputted, the clock unit 7 calculates the end time on the basis of the time when the voice recognition unit recognized input of the end term, the voice recognition unit 3 stores speech from the start time to the end time as recorded speech, analyzes the recorded speech.

JP2015-75792A discloses a report creation system utilizing a mobile terminal that is carried by a worker who performs maintenance/inspection work of a property. In this report creation system, maintenance/inspection results for each work target item of the property are consolidated by information based on speech input by the worker.
This report creation system automatically consolidates the information input by speech for each work target item of the property, and thus its use is limited to maintenance/inspection work of a property. Further, this system cannot convert arbitrary content into a report.


A liner running system for placing a liner in a well bore comprises a latch sleeve connected to the liner and a running tool releasably attached to the latch sleeve. The latch sleeve has a wall thickness substantially the same as the wall thickness of the liner, and the running tool has an outer diameter less than or equal to the inner diameter of the liner. The latch sleeve may be connected into the liner at any location along its length. The running tool is releasable from the latch sleeve via hydraulic actuation or mechanical actuation. Optionally, a swivel may be connected to the liner. The swivel has a wall thickness substantially the same as the wall thickness of the liner. The method for lowering the liner into the well bore via the liner running system comprises rotating the liner, pushing the liner, pulling the liner, or a combination thereof.

When drilling or completing wells that traverse earth formations, an upper portion of the primary well bore is drilled from the earth's surface to a selected depth, and then lined with a first section of pipe, commonly referred to as surface casing. The surface casing is then cemented into place in the well bore. The next succeeding section of the primary well bore is drilled to a selected depth below the surface casing and then lined with a string of pipe, commonly referred to as a liner. For each succeeding section of well bore that is drilled, a liner string is installed into the open borehole, below the surface casing or a previously installed liner string. During this process, each liner string may be cemented into place in the well bore.
Once the primary well bore is drilled and lined, a secondary well bore, such as a lateral well bore, for example, may be drilled and also lined with a liner. To perform a liner installation in either a primary or a secondary well bore, a running tool is releasably attached to a liner string. The running tool is connected to a work string or drill pipe that lowers the liner from the earth's surface into the open borehole below the surface casing or a previously installed liner string. The liner string may be rotated via the running tool to clear any obstructions in the borehole and to reduce friction as the liner string is lowered toward the bottom of the borehole.
Each liner string is connected at its upper end to a tubular liner hanger or another type of connection tubular, such as a lateral tube that extends between a primary well bore and a secondary well bore. The liner is lowered on the running tool via the work string until the liner hanger or connection tubular is adjacent to or near the lower end of the surface casing or a previously installed liner string. Then the liner hanger is set to engage the surrounding pipe wall and support the weight of the liner. Alternatively, the connection tubular is attached to the lower end of a previously installed casing or liner. Once the liner string is set, the liner may be cemented into place. The running tool is subsequently released from the liner and retrieved with the work string as it is withdrawn from the well bore.
Liner running tools conventionally include either hydraulic release means or mechanical release means. However, some liner running tools include both hydraulic and mechanical release means. Incorporating two different types of release means in a running tool is desirable given that trips into a well bore are expensive and time consuming. Thus, if the hydraulic release means fails, or if a liner must be reset, selective use of mechanical or hydraulic release means is desirable.
As described above, concentric liner strings are installed in the borehole as drilling progresses to increasing depths in a primary well bore or increasing lengths in a secondary well bore. Each new liner string must be run through the previously installed surface casing or liner string. Therefore, as successively smaller diameter liner strings are set, the flow area for the production of oil and gas is reduced. To maximize the production flow area, it is desirable to install a liner string with as large a diameter and length as possible so that the bottom of the formation can be reached with a comparatively larger diameter liner, thereby providing more flow area for the production of oil and gas.
However, traditional liner running tools have an outer diameter substantially the same size as the outer diameter of the liner string. Therefore, such running tools can only attach to the top of the liner string, and they act to "push" the liner string into the borehole. The longer the liner string, the more difficult it is for a traditional running tool to "push" the liner string into the borehole, especially in a lateral well bore. Therefore, it may be advantageous for a running tool to be releasably attachable to the liner string at any position along its length. Such a design would allow for the running tool to be connected near the lower end of the liner string, for example, so that a very long liner string may be "pulled" rather than "pushed" into an open borehole.
Further, some liner strings include features, such as slots or windows, which create structural weak points in the liner string. A running tool that could be attached to a liner string below a structural weak point, for example, would prevent stressing the weak point and buckling the liner string as it is being lowered into an open borehole. Therefore, a need exists for a liner running tool that may be releasably attached to a liner string at any location along its length.


An object analysis method performs object segmentation to generate segmentation confidence map and uses the segmentation results to generate robust object features. The robust object features are combined to create robust FOV summary features, robust sample summary features and robust assay summary features.

High content, quantitative analysis of microscopy images is an increasingly important tool for applications in drug discovery, basic research and medical diagnosis. We define image based, high content analysis to mean the measurement of multiple image parameters per cell or subcellular compartment or objects, across multiple cells in an image, or across multiple images. This could be done automatically in a high volume and high throughput manner or in a research setting that involves few cells or images in a semi-automatic fashion. High content analysis of these assays has only become practical in drug discovery and medical diagnosis in recent years, and is currently being adopted in basic research.
Prior to the advent of high content screening systems, prior art approaches in cell based screening only analyzed a single average fluorescent response of many hundreds of cells in a biological sample, usually contained in a microtiter well. A popular assay instrument that uses this approach is the Molecular Devices FLIPR (www.moleculardevices.com). High content screening tools in drug discovery have been deployed since the late 1990s. These individual cell based assays provide researchers with large amounts of biological and chemical information, and they offer important enhancements to information obtained through traditional high throughput screens. High content assays have to date been mostly deployed to screen chemical compounds against biological targets (usually receptors) genetically over-expressed in cell culture. More recently, high content assays have been increasingly adopted in target discovery; an important and popular application is RNA interference (RNAi) assays. The same imaging equipment and image informatics can be used in either case. High content analysis enables the measurement of complex and biologically important phenotypes that could not be measured in HTS, such as morphology changes, cellular differentiation, cytoskeletal changes, cell to cell interactions, chemotaxis and motility, and spatial distribution changes like receptor trafficking or complex formation.
Recently, high content analysis has become vital to cell culture automation, which has been identified as a critical bottleneck in both high content and high throughput screening. Here cell image analysis could be adapted to measure cells in microplates, count the cells, measure the confluence of cells, and the purity of cell culture (single or multiple clones). An example of this is a recent collaboration announced between MAIA Scientific and The Automation Partnership ("TAP Taps MAIA Scientific's Imaging System to Enable Automated Cell Culture for Well Plates" in Inside Bioassays Vol. 1(4) pg 1-5) to add Maia's image analysis software to the Cello automated cell culture system.
Chemical compound screening and RNAi based protein screening are accelerating the adoption of high content image based analysis in academic and basic research settings. Of course, microscopy has long been a benchtop tool for biologists, but until recently acquiring images using camera and analysis of those images has typically been low volume, low throughput, semi automatic with manual Region Of Interest (ROI) drawing and application of simple measurement tools included with standard digital microscopy software packages such as Universal Imaging's Metamorph, NIH Image, and MediaCybernetics' ImagePro. This appears to be changing as the NIH makes a strong push into chemical compound screening for academics. The Molecular Libraries and Molecular Imaging initiative (http://nihroadmap.nih.gov/molecularlibraries/index.asp) is a key component of the new NIH Roadmap (Zerhouni in Science Vol. 302(3) pg. 63-64 and 72, October 2003) and will offer public sector biomedical researchers access to small organic molecules which can be used as chemical probes to study cellular pathways in greater depth. It is intended for these assays to make use of high content and high throughput screening approaches, and NIH funding will likely favor researchers who adopt these types of tools. Probably a guiding case for the MLMI initiative, the NCI funded Harvard Institute for Chemistry and Cell Biology Initiative for Chemical Genetics (Stuart Schreiber: biology from a chemist's perspective in DDT Vol. 9(7) April 2004, pg. 299-303) has been using high content analysis of chemical compound screens for some time. They use chemicals in an analogous way to mutations, to dissect cellular pathways and identify previously unknown pathway components.
Very recently, RNAi has been validated as a platform technology for the analysis of protein function, and these assays benefit immensely from high content analysis to interpret the phenotypic changes of a sample subject to genetic perturbation (Carpenter, Sabatini, SYSTEMATIC GENOME-WIDE SCREENS OF GENE FUNCTION, in Genetics Vol. 5 pg. 11-22, January 2004). In the near future, genome wide screens will be commonplace. Several consortia (Netherlands Cancer Institute/Cancer Research UK, Vienna's Research Institute of Molecular Pathology/EMBL/Sanger Institute, Cold Spring Harbor Laboratories, and the RNAi consortium) have announced plans to make RNAi collections for the entire human genome. The Sloan-Kettering Institute and GE Healthcare have recently begun a collaboration to develop a technology capable of scanning the entire human genome in one day to analyze the function of each of the bodies 35,000 genes in a cellular process (see www.amersham.co.uk/investors/IR03/rep-4.html). This gene scanning technology will depend heavily on high content analysis software disclosed in "Harris et al. US Patent Application no. 2003/0036855 Method and Apparatus for Screening Chemical Compounds". Gene scanning will be made available to the broad academic community via a low-end hardware and optics platform that uses the same high content analysis software, a trend that indicates the growing importance of analytical software relative to hardware and optics platforms that are becoming commoditized.
There are many prior art approaches of cell analysis. "Lee, Shih-Jong J. U.S. Pat. No. 5,867,610 Method for Identifying Objects Using Data Processing Techniques, February 1999" discloses a method for the analysis of images of cervical Pap smear slides that enabled the first fully automated and FDA approved Pap smear screening device. In drug discovery, high content screening systems utilize advanced fluorescence light-microscopy and molecule specific fluorescent-protein tags to directly examine the physiology of fixed and living cells. Leading examples of state of the art devices are disclosed in "Harris et al. US Patent Application no. 2003/0036855 Method and Apparatus for Screening Chemical Compounds" and "Dunlay et al. U.S. Pat. No. 5,989,835 System for Cell Based Screening, November 1999".
The de facto standard for measuring assay quality in high throughput and high content screens is the z factor, disclosed in "Zhang et al, A Simple Statistical Parameter for Use in Evaluation and Validation of High Throughput Screening Assays, in Journal of Biomolecular Screening Vol. 4(2) pg. 67-73, 1999". Recently, it has been proposed that the Z factor also be used as a measure of quality for the new screens of RNAi induced phenotypes as well (Carpenter, Sabatini, SYSTEMATIC GENOME-WIDE SCREENS OF GENE FUNCTION, in Genetics Vol. 5 pg. 11-22, January 2004). It is reasonable to assume that the Z factor will see widespread use in academia as high throughput, high content assays are adopted.
The Z factor measures the assay signal window with a dimensionless parameter. The signal window can be thought of as the separation band between the distribution of test samples and that of control samples. This window is important to reduce false positive and false negative results. The Z factor is defined as:
  Z  =      1    -                  (                              3                                                                                σ              s                                +                      3                                                                                σ              c                                      )                                                  μ            s                    +                      μ            c                                      where σs and σc indicate the standard deviation of the sample and control populations respectively, and μs and μc indicate the mean of the sample and control populations respectively. As discussed in Zhang et al., the Z factor is sensitive to both data variability and the signal dynamic range. For example, as (3σs+3σc) approaches zero (very small standard deviations), or as |μs−μc| approaches infinity (large signal dynamic range), the Z-factor approaches 1, and the HTS assay approaches an ideal assay. Typically, an excellent assay is one that has a Z factor score greater than 0.5.
Assay development can be thought of as an exercise in optimization of many assay inputs to deliver the highest possible Z factor either by increasing signal range or reducing variation. There are many potential sources of variation, though scientists tend to focus on biological variation rather than instrument variation as that is what they can directly control. Sources of biological variation include subtle differences in cells resulting from cell culture variation, differences in DNA transfection across cells, variation in imaging probe titer and probe characteristics (such as rate of dissipation) across cells, errors in liquid handling, and poor cell adhesion. Furthermore, high content measurements can be confounded by compound related artifacts that can cause false positives and false negatives; such as fluorescent compounds, toxic compounds and rare morphological changes that affect the biological signal on which the assay is based.
Indeed, the evaluation of high content assay quality is fundamentally different than that of HTS assay quality because the sample unit is different. In HTS the sample is a single fluorescent measurement corresponding to microtiter well. In a high content assay, the sample is a biological object upon which a measurement or set of measurements, including combined and higher order measurements, are made using high content image analysis. There can be hundreds of objects in a FOV, and many FOVs per well, slide or cell array. Thus, high content analysis introduces a new source of variation into the measurement of assay quality: image analysis.
To date there has been no discussion in the literature or marketplace about how robust methods can be applied to high content analysis to both reduce measurement variation and increase the signal strength. It would be greatly beneficial to the field if robust methods could be deployed that yield a high quality assay while allowing the same or even more variation in assay inputs. This is possible in high volume, high throughput, microscopy image based assays because the high content image analysis plays a direct role in establishing both the signal dynamic range and the population variation.
Fundamentally, high content image analysis techniques can be used to reduce measurement variation at the sample level. Current state of the art approaches have in common the production of a binary mask. A binary mask image is a 1 bit image composed of ones (foreground) and zeros (background). The binary mask image corresponds to an input image of a high content assay wherein image segmentation has been applied. Image segmentation is the association of pixels to biological objects (e.g. cells or subcellular components). In the binary mask image the white areas (filled with ones) correspond to objects, and the black areas (filled with zeros) corresponds to the background. Object based measurements are carried out using the original input image within the region defined by the binary masks or their surrounding regions often subject to adjustments such as a correction for the non-uniform response of the imaging system across the field of view or transformation from intensity value to optical density. Common object based measurements include total intensity, average intensity, and standard deviation of intensity within the object region. Many other morphological features such as shape, texture and color measurements can also be made.
As described in "Harris et al. US Patent Application no. 2003/0036855 Method and Apparatus for Screening Chemical Compounds, the basic cell mask can be used to take measurements of nuclear and cytoplasmic activity. One example is for a two image fluorescent assay wherein one image corresponds to an emission filter channel that displays a Hoechst nuclear marker and a second image corresponding to a fluorescent reporter molecule describing some biological activity located in the cytoplasm. Object masks can be created by a simple threshold based segmentation algorithm applied to the Hoechst image, thus each object corresponds to the a cell nuclear region as the intensity in the Hoechst image displays only intensity located in the cell nucleus. An erosion image processing operation can be applied to these masks to create the nuclear mask. These masks can be used to measure the nuclear intensity in the corresponding regions of the Hoechst image. Next to measure cytoplasmic activity in the second image, a mask to represent the cytoplasm area must be created. To do this a dilation operation using preset parameters is applied to the original binary mask image, and areas that were one (1) in the original mask area are set to zero (0). The result is a donut shaped mask, these masks are used to measure cytoplasmic intensity in the corresponding regions of the second fluorescent image.
A similar method is disclosed in "Dunlay et al. U.S. Pat. No. 5,989,835 System for Cell Based Screening" and two examples of determining nuclear translocation of a DNA transcription factor are discussed. Firstly, an unstimulated cell with its nucleus labeled with a blue fluorophore and a transcription factor in the cytoplasm labeled with a green fluorophore. Secondly, the nuclear binary masks are created by performing cells segmentation on the fluorescent image corresponding to the blue fluorophore. The cytoplasm of the unstimulated cell imaged at a green wavelength. The nuclear mask is eroded (reduced) once to define a nuclear sampling region with minimal cytoplasmic distribution. The nucleus boundary is dilated (expanded) several times to form a ring that is 2-3 pixels wide that is used to define the cytoplasmic sampling region for the same cell. Using the nuclear sampling region and the cytoplasmic sampling region, data on nuclear translocation can be automatically analyzed by high content analysis on a cell by cell basis.
Binary mask based high content measurements introduce error into the assay at an early stage, in addition to instrument error such as focusing errors and variation in illumination. Types of measurement error are shown in FIG. 1A-4H. FIG. 1A-1D show errors in measurement on the nuclear image. The dark regions 104, 106 are the binary masks resulting from segmentation. The true nuclear regions 100, 102 are highlighted in checker patterns. Measurement errors result from segmentation errors that include over-segmentation (FIG. 1A), under-segmentation (FIG. 1B), missed segmentation (FIG. 1C) and overlapped segmentation (FIG. 1D). As described above, the nuclear masks 104, 106, 108 are used to derive cytoplasm rings 112, 114, 116 within which measurements are made on the cytoplasm regions 110, 118. FIGS. 1E-1H show how errors in measurements on the cytoplasm image accumulate from the initial segmentation errors made when creating the nuclear masks 104, 106, 108. The cytoplasmic rings 112, 114, 116 are shown in dark black overlain on the representation of the true nuclear 100, 102 (checker patterns) and cytoplasm regions (dotted patterns). As disclosed above, cytoplasmic region measurements are meant to measure the fluorescent activity of fluorophores in the cytoplasm, however types of common measurement errors include measuring both the true cytoplasm and true background intensities within the cytoplasm ring region 112 (FIG. 1E), measuring intensities corresponding to true cytoplasm, true background and true nuclear regions within the cytoplasm ring region 114 (FIG. 1F), missing the object altogether, and the cytoplasm ring region 116 measuring the cytoplasm intensity of two cells and treating it as one (FIG. 1H). This error is again accumulated and undermines derived measurements such as the standard deviation of intensity, the ratio of cytoplasmic to nuclear intensity, etc.
Similar error is accumulated in time lapse images when objects are not perfectly aligned from frame to frame. Error is introduced when the nuclear object reference mask and the true nuclear object shift over time. As the nucleus shifts from image frame to image frame, the measurement region corresponding to the initial binary mask increasingly includes background fluorescence in its measurement.
These fundamental errors in object segmentation and measurement are propagated throughout the assay's statistics resulting in higher assay variability and reduced signal dynamic range. Additional variation is introduced by instrument and biological variation. It is clear then that there is a need for robust methods of high content analysis that allow for a more accurate segmentation result, and more specific and sensitive measurements with high repeatability. These robust measurements are needed not only at the individual object level, but also at the FOV level, the sample level (usually corresponding but not limited to a microtiter plate well or slide bound tissue specimen or micro tissue array) and the assay level.

Method for robust analysis of biological activity in microscopy images