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Fly fishing for small stream trout in the WV mountains. All dry fly fishing, all C&R. Cameras used were a Canon HG10 and a GoPro Hero HD w/Blurfix flat lens modification. This was all self-filmed.
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McLaren to launch MP4-28 on January 31
McLaren will launch its 2013 car on the last day of January, the famous British team has announced.
A media statement issued by the Woking based outfit offered no further details except to say the MP4-28 unveiling "will take place in the UK".
It is likely most other teams will also reveal their new contenders around the same period, immediately before the three-test winter pre-season period begins in Spain in early February.
We reported Peter Sauber's news recently that the Swiss team's new car, the C32, will be launched on the first day of the first test at Jerez, on February 5.
Force India's car will be publicly shown at Silverstone, the day after McLaren's.
And Italy's La Gazzetta dello Sport said the new Ferrari will probably be launched on either 1 or 2 February.
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KINGSTON, R.I. -- Oct. 29, 2002 -- University of Rhode Island senior Kathleen McPherson has always had an interest in birds, which is why her research project is no surprise. McPherson is studying the nutritional habits of migratory birds as part of the Universitys Coastal Fellows Program.
"I have always had birds as pets," said the North Smithfield resident. "I think that helped to increase my interest in them, especially when my Cockatiels had two clutches of young. It was a great learning experience to see them raise their young."
According to McPherson, migratory birds change their diet with the seasons.
"Migratory birds eat insects, which are high in protein and fat, during the summer. During migration in the fall, the birds eat fruits, which are high in sugars or lipids," said McPherson. "No one really knows why they do this and thats the question Im trying to answer."
Over the last year, she has collected nine species of fruit from Block Island, a common stopover spot for migratory birds. "For this project, I am not studying the birds themselves, I am only studying their diets
the fruit," said McPherson.
In order to determine why the birds change their diets according to season, McPherson said, she had to first determine the energy-to-protein ratio of the fruit. A low energy-to-protein ratio helps the birds satisfy their protein requirements.
"Birds, like most animals, eat to satisfy their energy requirements rather than for particular nutrient requirements. Birds will eat fruit with a relatively high energy-to-protein ratio, and so, ingest less protein compared to a fruit with a relatively low energy-to-protein ratio," said McPherson, who discovered that the fruit of viburnum bushes give birds the most nutritional value due to its high energy and fat content.
McPherson is still analyzing the data she collected from her research with the assistance of URI graduate students Barbara Pierce and David Podlesack and URI Assistant Professor Scott McWilliams.
The toughest part of the project was separating the seeds from the fruits, joked McPherson. "I spent a long time picking the seeds out of the fruit, when I could have just freeze-dried the fruit. That would have been much easier."
Her research was funded by the National Science Foundation through the URI Coastal Fellows Program, a unique program designed to involve undergraduate students in addressing current environmental problems. Now in its seventh year, the Coastal Fellows Program teams students with URI faculty, research staff and graduate students to help undergraduates gain skills that will ensure their future success.
"I learned a lot from the Coastal Fellows Program," said McPherson. "Students get hands-on experience analyzing data, using trial and error and gaining experience in the field. With Coastal Fellows, students get a taste of real research, not just lab work."
After graduation, McPherson wants to travel the country for a year doing fieldwork. "Im interested in studying birds outside of New England, especially in the west. It would be interesting to study the migratory patterns of birds elsewhere in the world."
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text: Bookmarks Testicular self-examPrint-Friendly Previous imageEnlargeCloseNext image / Previous imageEnlargeCloseNext image / NavigatorTesticular self-exam is an examination of the testicles that you do on yourself.TesticlesThe testes are two egg-shaped male reproductive organs located in the scrotum. They produce sperm and the male hormone, testosterone. Related topics...ImageRead Article Now Book Mark Article The testicles (also called the testes) are the male reproductive organs that produce sperm and the hormone testosterone. They are located in the scrotum under the penis. You can do this test during or after a shower. This way, the scrotal skin is warm and relaxed. It is best to do the test while standing.Gently feel your scrotal sac to locate a testicle. Use one hand to stabilize the testicle. Use your fingers and thumb on the other hand to firmly but gently feel the testicle. Feel the entire surface. Check the other testicle in the same way.I Would Like to Learn About:Expand All Collapse All How the Test is Performed The testicles (also called the testes) are the male reproductive organs that produce sperm and the hormone testosterone. They are located in the scrotum under the penis. You can do this test during or after a shower. This way, the scrotal skin is warm and relaxed. It is best to do the test while standing.Gently feel your scrotal sac to locate a testicle. Use one hand to stabilize the testicle. Use your fingers and thumb on the other hand to firmly but gently feel the testicle. Feel the entire surface. Check the other testicle in the same way. Why the Test is Performed A testicular self-exam may be done to check for testicular cancer.Testicular cancerTesticular cancer is cancer that starts in the testicles, the male reproductive glands located in the scrotum.ImageRead Article Now Book Mark Article Testicles have blood vessels and other structures that can make the exam confusing. If you notice any lumps or changes in a testicle, contact your health care provider right away.Doctors may recommend that you do a testicular self-exam every month if you have any of the following risk factors:Family history of testicular cancer Past testicular tumor Undescended testicleUndescended testicleUndescended testicle occurs when one or both testicles fail to move into the scrotum before birth.ImageRead Article Now Book Mark Article Some, but not all expert organizations, recommend self-exam if you are a teenager or young adult (to about 35 years old). It is not clear that performing this exam when you have no symptoms would decrease the risk of dying from testicular cancer. Normal Results Each testicle should feel firm, but not rock hard. One testicle may be lower or slightly larger than the other.Talk to your health care provider if you have questions. What Abnormal Results Mean If you find a small, hard lump (like a pea), have an enlarged testicle, or notice any other differences that do not seem normal, see your health care provider right away.Call your health care provider if:You cannot find one or both testicles -- the testicles may not have descended properly in the scrotum There is a soft collection of thin tubes above the testicle -- this may be a collection of widened veins (varicocele) VaricoceleA varicocele is the swelling of the veins inside the scrotum. These veins are found along the cord that holds up a man's testicles (spermatic cord)....ImageRead Article Now Book Mark Article You have pain or swelling in the scrotum -- this may be an infection or a fluid-filled sac (hydrocele) causing a blockage of blood flow to the areaHydroceleA hydrocele is a fluid-filled sack in the scrotum.ImageRead Article Now Book Mark Article Sudden, severe (acute) pain in the scrotum or testicle that lasts for more than a few minutes is an emergency. If you have this type of pain, seek medical attention right away.A lump in the testicle is often the first sign of testicular cancer. If you find a lump, see a health care provider right away. Most testicular cancers are very treatable. Keep in mind that some cases of testicular cancer do not show symptoms until they reach an advanced stage.Related Information TestesTestosteroneUndescended testic...Testicular cancer...VaricoceleHydrocele Open ReferencesReferencesNational Cancer Institute: Physician Data Inquiry (PDQ®) Testicular Cancer Screening. Bethesda, MD. Date last modified: July 19, 2012. Accessed January 7, 2013. Available at: //www.cancer.gov/cancertopics/pdq/screening/testicular/Patient/page3American Cancer Society. Testicular self-exam. Accessed August 14, 2013. Available at: //www.cancer.org/cancer/testicularcancer/moreinformation/doihavetesticularcancer/do-i-have-testicular-cancer-self-examMontgomery JS, Bloom DA. The diagnosis and management of scrotal masses. Med Clin North Am. 2011;235-244.U.S. Preventive Services Task Force. Screening for testicular cancer. U.S. Preventive Services Task Force reaffirmation recommendation statement. Ann Intern Med. 2011;154:483-486.BACK TO TOP Review Date: 12/27/2013 Reviewed By: Louis S. Liou, MD, PhD, Chief of Urology, Cambridge Health Alliance, Visiting Assistant Professor of Surgery, Harvard Medical School. Also reviewed by David Zieve, MD, MHA, Bethanne Black, and the A.D.A.M. Editorial team. The information provided herein should not be used during any medical emergency or for the diagnosis or treatment of any medical condition. A licensed medical professional should be consulted for diagnosis and treatment of any and all medical conditions. Call 911 for all medical emergencies. Links to other sites are provided for information only -- they do not constitute endorsements of those other sites. © 1997- A.D.A.M. content is best viewed in IE9 or above, Firefox and Google Chrome browser.Content is best viewed in IE9 or above, Firefox and Google Chrome browser.


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The city of Logan has moved another step forward in paving the way for a new walking bridge.
Street Commissioner Kevin Marcum said his department has received 65 tons of asphalt patch to be put down around the town. Marcum said a new drain pipe had been put in on Park Avenue.
"We also put down 230 feet of rubber on the walking bridge in Central City to cover the small holes that had been caused due to rust," Marcum added. "This is a temporary fix until the new bridge construction can begin. The belt we used was donated so we were not out much on the expense of the repair." He said the Logan Street Department had also been kept busy following the storm due to several trees which had fallen.
The new bridge project was a major topic of discussion Tuesday night. The council approved a resolution to repair the Central City Walking Bridge with a $34,000 loan from Logan Bank and Trust and $34,000 each from the Logan Board of Education and the Logan County Commission.
"We need that money to get a $500,000 grant from the Department of Transportation," the Mayor said, noting the terms of loan are 5.9 percent. "If we can get the approval for this, we hope to get the ball rolling and that by August we can sign the actual contract so the work can begin. Sometimes with these grants there are a lot of things that have to be done and approved first that are very tiring and time consuming but we feel this will be worth the effort."
• In other Logan news:
*Fire Chief Scott Beckett said his department had been busy in June and July with 82 calls, including 30 calls that came in during the extreme summer storm. Beckett said there were two house fires, a train hitting a car and other events that made it "an eventful week."
• *John Allen spoke with the council about the proposal to lease a postage and mailing machine for $330 a month. He demonstrated the device which weighs mail up to 30 pounds and packages items of all sizes. It also prints postage which can be affixed to the box and thanks to an internet link up changes in postage from the Post Office are made instantly.
"It is the same machine they use in Boone County's Courthouse," Allen said, noting that the machines can save both time and money thanks to a 5 percent discount on Priority Mail. Councilman Jay Mullins said it might also save time and papercuts.
• Council approved a rotating line of credit for $25,000 from Logan Bank and Trust. Mayor Nolletti noted the town does not have to use the offered loan, but added it was nice to know the money would be there if needed. One person at the meeting said it was a better idea to do business with LB&T than to take a fire truck to the Pawn Stars for a quick loan.


text:
A bio-separation system using an efficient, compact, portable, interchangeable, reusable, recyclable, multi-channel cartridge, has integrated pre-aligned optics and an integrated reagent reservoir. The cartridge supports, for example, multiple capillaries for CE separation. An integrated reservoir containing a separation support medium (e.g., a gel buffer) is common to all capillaries. The chemistry of the medium and the characteristics of the capillaries (e.g., capillary size, coating and length) are defined for each cartridge. Different cartridges can be easily interchanged in the bio-separation system to suit the particular sample based separation. The reservoir is coupled to an air pressure pump that pressurizes the gel reservoir to purge and fill the capillaries with buffer as the separation support medium. In another aspect of the present invention, optics requiring fine alignment with respect to the detection zones (such as fiber optics for directing incident radiation and/or radiation emissions) are integrated into the cartridge.
1. A multi-channel cartridge for bio-separation, comprising:
a body;
a plurality of capillary separation channels for analytes, the capillary separation channels are defined and isolated from one another by physical walls in the body, wherein the analytes are confined to migrate within boundary of the physical walls;
a chamber in the body defining a reservoir in fluid flow communication common with the capillary separation channels, said chamber containing a separation support medium which is sealed from leakage when the cartridge is handled in any orientation; and
built-in optics integrally supported within the body, the built-in optics being aligned with respect to a section of each capillary separation channel to define a detection zone for at least one of incident radiation and radiation output.
2. The multi-channel cartridge as in
3. The multi-channel cartridge as in
4. The multi-channel cartridge as in
5. The multi-channel cartridge as in
6. The multi-channel cartridge as in
7. The multi-channel cartridge as in
8. The multi-channel cartridge as in
9. The multi-channel cartridge as in
10. The multi-channel cartridge as in
11. The multi-channel cartridge as in
12. The multi-channel cartridge as in
13. The multi-channel cartridge as in
14. A bio-separation system, comprising:
a base;
a multi-channel cartridge for bio-separation supported on the base, comprising:
a body;
a plurality of separation channels, the capillary separation channels are defined and isolated from one another by physical walls in the body, each separation channel defining a detection zone, wherein analytes are confined to migrate within boundary of the physical walls;
a chamber in the body defining a reservoir in fluid flow communication common with the separation channels, said chamber containing a separation support medium which is sealed from leakage when the cartridge is handled in any orientation;
built-in optics integrally supported within the body, the built-in optics being aligned with respect to a section of each capillary separation channel to define a detection zone for at least one of incident radiation and radiation output;
a positioning mechanism positioning a container of samples supported in a horizontal plane with respect to the multi-channel cartridge such that the separation uliariueb arc in fluid communication with the container;
a separation mechanism effecting bio-separation of the samples along the separation channels; and
a controller controlling operations of the positioning mechanism and separation mechanism.
15. The bio-separation system as in
a radiation source directing radiation at the detection zone; and
a detector detecting radiation from the detection zone.
16. The bio-separation system as in
17. The bio-separation system as in
18. The bio-separation system as in
19. A bio-separation system, comprising:
a container supporting samples in a horizontal plane;
a multi-channel cartridge for bio-separation, comprising:
a body;
a plurality of separation channels, the capillary separation channels are defined and isolated from one another by physical walls in the body, each separation channel having a detection tone, wherein analytes are confined to migrate within boundary of the physical wails;
a chamber in the body defining a reservoir in fluid flow communicating common with the separation channels, said chamber containing a separation support medium which is sealed from leakage when the cartridge is handled in any orientation; and
a positioning mechanism, positioning the container of samples with respect to the multi-channel cartridge such that the separation channels are in fluid communication with the container; and
wherein the multi-channel cartridge further comprises built-in optics integrally supported within the body, which is aligned with respect to the detection zone for at least one of incident radiation and radiation output, and an electrode electrically coupled to the reservoir,
a controller controlling operations of the positioning mechanism.
20. The bio-separation system as in
a radiation source directing radiation at the detection zone; and
a detector detecting radiation from the detection zone.
21. The bio-separation system as in
22. The bio-separation system as in
This application claims the priority of U.S. Provisional Patent Application No. 60/264,605, filed on Jan. 26, 2001.
Reference is made to U.S. patent application Ser. No. 10/060,052 entitled "Optical Detection In A Multi-Channel Bio-Separation System", concurrently filed on Jan. 28, 2002, now U.S. Pat. No. 6,828,567, which is commonly assigned to BioCal Technology, Inc., the assignee of the present invention, and which is fully incorporated by reference herein.
1. Field of the Invention
The present invention relates to bio-separation, and more particularly a portable cartridge for supporting multi-separation columns with integrated detection optics and reagent reservoir and a bio-separation system incorporating the cartridge.
2. Description of Related Art
Bioanalysis, such as DNA analysis, is rapidly making the transition from a purely scientific quest for accuracy to a routine procedure with increased, proven dependability. Medical researchers, pharmacologists, and forensic investigators all use DNA analysis in the pursuit of their tasks. Yet due to the complexity of the equipment that detects and measures DNA samples and the difficulty in preparing the samples, the existing DNA analysis procedures are often time-consuming and expensive. It is therefore desirable to reduce the size, number of parts, and cost of equipment, to ease sample handling during the process, and in general, to have a simplified, low cost, high sensitivity detector.
One type of DNA analysis instrument separates DNA molecules by relying on electrophoresis. Electrophoresis techniques could be used to separate fragments of DNA for genotyping applications, including human identity testing, expression analysis, pathogen detection, mutation detection, and pharmacogenetics studies. The term electrophoresis refers to the movement of a charged molecule under the influence of an electric field. Electrophoresis can be used to separate molecules that have equivalent charge-to-mass ratios but different masses. DNA fragments are one example of such molecules.
There are a variety of commercially available instruments applying electrophoresis to analyze DNA samples. One such type is a multi-lane slab gel electrophoresis instrument, which as the name suggests, uses a slab of gel on which DNA samples are placed. Electric charges are applied across the gel slab, which cause the DNA sample to be separated into DNA fragments of different masses.
Another type of electrophoresis instrument is the capillary electrophoresis (CE) instrument. By applying electrophoresis in a fused silica capillary column carrying a buffer solution, the sample size requirement is significantly smaller and the speed of separation and resolution can be increased multiple times compared to the slab gel-electrophoreses method. These DNA fragments in CE are often detected by directing light through the capillary wall, at the components separating from the sample that has been tagged with a fluorescence material, and detecting the fluorescence emissions induced by the incident light. The intensities of the emission are representative of the concentration, amount and/or size of the components of the sample. In the past, Laser-induced fluorescence (LIF) detection methods had been developed for CE instruments. Fluorescence detection is often the detection method of choice in the fields of genomics and proteomics because of its outstanding sensitivity compared to other detection methods.
Some of the challenges in designing CE-based instruments and CE analysis protocols relates to sample detection techniques. In the case of fluorescence detection, considerable design considerations had been given to, for example, radiation source, optical detection, sensitivity and reliability of the detection, cost and reliability of the structure of the detection optics. In the past, a relatively high power light source is required, such as a Laser. When light is directed through the capillary wall at the separated sample components in the capillary bore, light scatters at the outside capillary wall/air interface and the inside capillary wall/buffer interface (Raman scattering), which obscures or corrupts the fluorescence emission intensity. Similarly, fluorescence emissions scatter at the wall interfaces. In the past, various techniques were developed for more completely collecting the fluorescence emissions to improve signal intensity and hence detection sensitivity. These techniques involve additional moving and non-moving components that add to the relative complexity and cost of the detection setup.
The design limitations of prior art electrophoresis instruments are exacerbated in the development of multi-capillary CE-based instruments. For example, confocal scanning laser induced fluorescence (LIF) detection has been adopted in multi-capillary electrophoresis systems. The scanning confocal detection relies on a scanning optical system. The use of moving parts is not ideal when taking simplicity, robustness, and lower cost of the instrument into consideration. Also, the shallow focal depth of the microscope objective for the confocal detector puts severe demands on the mechanical and optical component tolerances. Further, the optical scanning method generally involves a longer duty cycle per capillary. Thus, should the instrument be scaled up in order to generate higher throughput, the sensitivity of the system may be compromised. Also, another detection method is Sheath Flow detection. The main drawback of the sheath flow detector is the highly sophisticated flow system needed to ensure a reliable sheath flow with minimum optical cross talk between the channels. Extreme demands are put on the optical and mechanical component tolerances in order to meet the robustness demands of end-users. The sensitivity of the device is very good, but it is not obvious that this principle of fluorescence detection is suited for a high-throughput, yet low cost, DNA analysis.
Additional challenges in designing multi-capillary CE-based instruments related to the support of the capillaries. U.S. Pat. No. 5,198,091 to Burolla et al. describes a capillary cartridge for electrophoresis that employs a long length of capillary arrays. This patent may include a hollow space defined about the capillary for circulating coolant fluid but it does not include a reservoir as an integrated part of the cartridge. U.S. Pat. No. 5,413,686 to Klein et al. describes an automated multi-channel capillary electrophoresis analyzer including a plurality of capillaries. Reservoirs are shown in the analyzing apparatus, but they are multiple reservoirs and they are separated from the capillaries, not integrated into a capillary support. Detection optics are also shown in the apparatus, but they are not integrated into a compact capillary support. U.S. Pat. No. 5,338,427 to Shartle et al. describes a single use separation cartridge for a capillary electrophoresis instrument, in which capillary tubes are horizontally disposed in a coplanar array. The single use separation cartridge replaces large reagent reservoirs with hemispherical drops of reagent.
Also, current systems for gel buffer chemistry do not allow use of the CE instrument that is specific with applications. In other words, current CE instruments require matching the capillary (with different coatings and column sizes) with the buffer reagent for different separation applications (different types, speeds, resolutions).
The present invention provides a bio-separation system that uses an efficient, compact, simplified, portable, interchangeable, reusable, low cost, recyclable, easy to assemble multi-channel cartridge with no moving parts for bio-separation, which has integrated pre-aligned optics and an integrated reagent reservoir. The cartridge supports, for example, multiple capillaries for CE separation. The integrated reservoir containing a separation support medium (e.g., a gel buffer) is common to all capillaries. The chemistry of the medium and the characteristics of the capillaries (e.g., capillary size, coating and length) are defined for each cartridge. Different cartridges can be easily interchanged in the bio-separation system to suit the particular sample based separation. The reservoir is coupled to an air pressure pump that pressurizes the gel reservoir to purge and fill the capillaries with buffer as the separation support medium. In another aspect of the present invention, optics requiring fine alignment with respect to the detection zones (such as fiber optics for directing incident radiation or radiation emissions) are integrated into the cartridge.
In one aspect of the present invention, the cartridge supports multiple capillaries for CE separation. The cartridge includes assembled body parts, excitation fibers, capillaries, electrodes, a buffer/gel reservoir, and integrated optics for external radiation input. The reservoir is equipped with a single electrode common to all capillaries.
In another aspect of the present invention, optics are integrated into the cartridge. According to an embodiment of the present invention, the optical excitation system is integrated with the cartridge. The excitation system includes directing excitation light by excitation fibers to detection zone by coupling LEDs with micro-ball lenses. The excitation fibers are routed to a V-groove assembly adjacent to each capillary. According to another embodiment of the present invention, the optical detection system is engaged with the cartridge by a shutter mechanism. The detection optics for each of the capillaries, or the detection array, is coupled to a single photo-multiplier tube. The detection array includes collimating the emission light from the detection zone by using micro-ball lenses and detection fibers.
In a further aspect of the present invention, the present invention provides a bio-separation instrument that incorporates the multi-channel bio-separation cartridge of the present invention.
For a fuller understanding of the nature and advantages of the invention, as well as the preferred mode of use, reference should be made to the following detailed description read in conjunction with the accompanying drawings. In the following drawings, like reference numerals designate like or similar parts throughout the drawings.
This invention is described below in reference to various embodiments with reference to the figures. While this invention is described in terms of the best mode for achieving this invention's objectives, it will be appreciated by those skilled in the art that variations may be accomplished in view of these teachings without deviating from the spirit or scope of the invention.
The present invention is directed to a novel CE system and novel cartridge configuration in which incident radiation (e.g., from a laser or LED source) for detection of separated analytes is directed through the boundary walls of the detection zone or the separation column. For purpose of illustrating the principles of the present invention and not limitation, the present invention is described by reference to embodiments directed to capillary electrophoresis and radiation induced fluorescence.
Referring to
One end of the capillary column 22 is submerged in a reservoir 28 of running buffer/gel 34. The other end of the capillary column 22 is coupled to the sample vial 26. It is understood that the detection configurations shown in the other embodiments can be equally implemented in a system similar to the CE system 20. Also, the separation channel 36 may be one straight capillary or micro-channel with a section of the detection window closest to the gel-reservoir at the exit end being the detection zone, which is the current preferred mode of our invention. A radiation detector 24 is positioned outside a transparent section of the capillary walls at detection zone 30. An excitation fiber 16 extends from a radiation source 18 (e.g., LED or laser) and is directed at the detection zone 30 outside the walls of the column. Electrodes 12 and 14, that are part of the cartridge assembly are coupled to the buffer reservoirs 26 and gel reservoir 28 to complete the electrophoresis path.
For the sake of completeness, it is sufficient to briefly mention the operation of the CE system 200. In operation, a prepared biological sample (e.g., a DNA sample), direct from Polymerase Chain Reaction (PCR) machine is introduced into the far end of the capillary column away from the detection zone by any of a number of ways that is not part of the present invention (e.g., electrokinetic injection from a sample reservoir or physical pressure injection using a syringe pump). The sample binds to the fluorophore.
When a DC potential (e.g., 1-30 KV) is applied between electrodes 12 and 14, the sample migrates under the applied electric potential along the separation channel 36 (e.g. DNA that is negatively charged travels through the sieving gel with an integrated dye matrix/fluorophore toward a positive electrode as shown in
When the sample reaches the detection zone, excitation radiation is directed via the excitation fiber 16 at the detection zone. The sample components fluoresce with intensities proportional to the concentrations of the respective sample components (proportional to the amount of fluorescent tag material). The detector 24 detects the intensities of the emitted fluorescence at a wavelength different from that of the incident radiation. The detected emitted radiation may be analyzed by known methods. For an automated system, a controller 32 controls the operations of the CE system 200.
The multi-channel capillary array includes twelve detection zones 30 defined by micro-channels 36 in cartridge body (also see
The cartridge 100 described in greater details below. Briefly, the cartridge 100 includes a twelve-channel fused silica capillary array that is used for separation and detection of the samples as part of a disposable and/or portable, interchangeable cartridge assembly 100. The multi-channel capillary array includes twelve detection zones defined by micro-channels in the cartridge 100. The multi-channel cartridge 100 shown in
In accordance with one embodiment of the present invention, the block diagram of the controller 32 for the CE system 200 is shown in
The components of the controller 32, with the exception of the PC 218, may be packaged as an electronic board 64 (
Injection of the samples is achieved by electrokinetic methods. The high voltage power supply 76 is used to deliver 0-to-20 KV of electrical field to the gelfilled capillaries for the electrokinetic injection and separations of DNA fragments. Each of the 12-LED's broad band light energy (FVHM=47 nm) is relayed by individual light transmitting optical fibers (multi-mode silica or plastic 200 micron Core fibers, 0.22 N.A.) to each of the capillary's detection zone inside the cartridge 100 for the excitation of the separated DNA fragments.
In operation, the sample handling tray transport mechanism 80, with a 96-well plate (8�12), is used to introduce the amplified DNA samples (or analytes) to each microbore channel 36. Inside the micro-channels 36 are Polyimide coated or glass capillary tubings 22 of smaller inner diameter (25-100 μm) used as separation columns. The X-Z transport mechanism 80 indexes a row of sample carrying wells under the row of capillary tips and dip the tips into the well. By applying a voltage, electrokinetic injection moves a known amount of the DNA sample to the beginning of the separation column 140. After injection, the DNA samples from sample tray 72 maybe replaced with a running buffer from tray 70. Alternatively, after injection, the transport mechanism 80 may index to move a row of 12 wells containing buffer solution into position under the cartridge to replace the twelve wells containing DNA samples. By applying high voltage across the total length of the capillary separation channel and the micro-channel 36, separation of the DNA sample into DNA fragments is achieved. Up to 1000 V/cm (typically 300 V/cm) of high voltage is applied, which provides fast separations of less than 10 minutes along the entire length of the separation channel. The total separation length is about 12.5 cm up to the detection zone. The separation capillary length inserted inside the micro-channel is about 6.5 cm. High voltage is applied to a total active length of 16-17 cm, which could be the length from the bottom to the top of one single capillary with 75 micron I.D. inside the gel-reservoir as a single separation and detection capillary. During electrophoresis, the rate at which the DNA fragments move through the sieving gel is inversely proportional to their mass; i.e., lighter (or smaller) DNA fragments move more quickly than heavier (or larger) ones. As the fragments approach the end of the separation column 22 and enter into the detection zone 30, the excitation light energy from each of the twelve LEDs (not shown) is delivered by individual light transmitting optical fibers from outside the detection window, illuminating the migrating DNA fragments from sample tray 72. As the DNA fragments move through the sieving gel, or linear polymer solution (e.g., 25 mM Mops-Tris pH 7.55, as referenced in "Pace Setter", Vol. 3, Issue 1, Apr. 1999), a DNA intercalating dye (Ethidium Bromide) within the sieving gel allows the migrating DNA fragments to be detected. Experiments have shown that detection sensitivities of 100 ng/ml (0.02 ng of the Haelli digest φX174 DNA test mix) are achievable, which is several orders of magnitude better than conventional slab gel electrophoresis devices using the same intercalating dye. As the twelve LEDs are time-multiplexed (with sampling frequency of 10-100 Hz), twelve emission signals coupled to twelve emission detection fibers will reach the single PMT in a time-staggered manner by a single fiber-bundle assembly.
To prepare for the next run with a different sample, the old gel from the previous run is purged from the capillaries by pressuring the reservoir to refill the capillaries with fresh gel. The trays 70 and 72 carries cleaning solutions, waste collection, and samples. The purged gel is collected by one of the trays 70 and 72 by positioning the tips of the capillaries at a row of waste collecting wells in one of the trays. The tips of the capillaries may be cleaned with water or a cleaning solution by positioning and dipping the tips of the capillaries in such solution in the appropriate tray wells. When the capillaries are refilled and ready for the next run, the tips of the capillary are dipped into the samples by repositioning the trays 70 and 72. The above mentioned sequence of process may be programmed as one of the automated functions of the controller 32.
It is noted that because the sample analytes that flowed to the gel reservoir at the exits of the capillaries are in such small amount and volume concentration compared to the volume of the reservoir, and that the analytes are expected to be mixed within the gel reservoir, there will only be a negligible trace of analytes from past runs in the reservoir, and that will be evenly distributed in the gel that refills the capillaries for the next run. Any noise from this negligible trace would be relatively small background noise that can be easily removed from the detected signal in the data analysis.
After the mid-section body 120 of the cartridge is mounted onto the lower-section body 110, as shown in
The reservoir 130 containing the gel is sealed, such as hermetically sealed at the body of the cartridge, which allows the cartridge to be handjed by holding it in any orientation without leakage of the gel. (There is negligible leakage or exposure at the capillary tips because of surface tension and high viscosity within the inicrobore of the capillaries.) The cartridge 100 has a rubber septum (not seen) that is pierced by an instrument-mounted needle (or any sharp object) that provides air pressure from the pump 78 into the cartridge. This allows air pressure to fill the capillaries with the gel/buffer solution after each separation run, and to purge the old gel from the previous run in the process. This approach assures the proper containment of the gel inside the cartridge reservoir; it also provides a simple and reliable means of accessing the gel reservoir and of providing enough air pressure for the gel to fill up the capillaries prior to applying high voltage to effect CE separation.
The cartridge 100 also has detection optic ports 161 through which detector probes 170 (
The last stage of assembling the cartridge is shown in
The cartridge has alignment features to be easily aligned to the micro-optical detection module inside the instrument 200. The optical detection array 170 and LED array 184 are all spring loaded, which provides independently compliant forces to each lens barrel assembly 188 (i.e. LED or fiber ferrule) for a reliable and repeatable alignment to the cartridge. The cartridge has all the proper conical type features (i.e., conical lens seating 186) to accept the spring and the spring loaded arrays from the instrument, as will be described in greater detail below.
A closer look at section A in
Two excitation fibers 116 for two wavelengths (for each capillary) are integrated inside the cartridge 100, with fixed alignment, at close proximity to the capillary detection zone 155. These two excitation fibers 116 are coupled to two LEDs 184 (e.g., two different colors: 526 nm and 473 nm) when the cartridge is installed inside the CE instrument 200 (i.e., DNA Analyzer). Two colors can be separated and detected by two-color emission filters at the detection module (PMT module 178). The cartridge 100 can have single color capabilities for DNA fragment analysis applications and also can be upgraded to have two-color detecting capabilities for other applications. Reference is made to U.S. Provisional Application No. 60/348,034 entitled "A Portable Multi-color Multiplexed Analysis Electrophoretic Device," filed on Oct. 19, 2001, which is commonly assigned to BioCal Technology, Inc., the assignee of the present invention, and which is fully incorporated by reference herein.
U.S. patent application Ser. No. 10/060,052 entitled Optical Detection in A Multi-Channel Bio-Separation System, concurrently filed on Jan. 28, 2002, which is assigned to BioCal Technology, Inc., the assignee of the present invention, and which had been fully incorporated by reference herein, is more specifically directed to the time staggered/multiplexed detection scheme that can be adopted in the CE system 200 in which the cartridge 100 is designed to be used.
A closer look at section B in
When the excitation light is directed at the detection zone 155 (also see
It is further noted that the detection zone is not necessarily a well-defined zone with well-defined boundaries, due to the nature of the substance, the incident radiation, and the fluorescence emissions. It is generally a zone in which light from the excitation fiber is directed to cause fluorescence emissions and the detection optics is aimed to capture part of such fluorescence emissions. Light from the excitation fiber may cause fluorescence emissions outside the detection zone, and some of the emissions from within the zone may not be detected by the detection optics. The closer the excitation fiber is to the detection zone or the higher the power density of excitation light, the stronger the collected emission signals are.
In the multi-capillary CE device of the present invention, the fluorescence excitation light sources may be super bright blue or green LEDs. The attractive features of LEDs as light sources are their low cost, small size, long lifetime, good intensity and stability resulting in low noise, and the possibility of direct electronic modulation of the intensity. The LEDs contemplated in this invention are based on InGaN material technology (e.g., HLMP-CB 15 and HLMP-CM15 from Agilent) with an average light output power of 2.5-3 mW. The spectral characteristics with its peak wavelength and halfwidth (nm) of the InGaN LEDs indicate that these LEDs can be used for excitation of fluorescence with excitation spectra in the range of 440 to 570 nm (e.g., fluorescin, rhodamine, Etidium Bromide, thiazol orange) and for frequency in the range of 1 Hz to 100 MHz. Since the response time of these LEDs are very high (at a few hundred nanoseconds), they can be pulsed at greater forward currents, up to 100 mA in pulsed mode operation, to obtain high radiant peaks. Pulsed operation of LEDs can typically be achieved by the transistor drive circuits. Significantly higher peak LED light output can be realized from large drive current pulses at low duty cycles (i.e., 5%, 10%, 25% or 50%) than DC operation.
Different color LEDs (i.e., blue or green LEDs) could be used as excitation sources for excitation of different fluorophores (different applications). The preferred embodiment uses LEDs in wavelength ranges of 500-600 nm, and specifically at 524 nm. A second LED module, or a second color LED, could be added to the current design for a dual-wavelength detection device either bringing two wavelengths to the micro-channel using one or two fibers. The current detection/separation platform could be expanded with dual LED modules by having excitation and collection optics with a second PMT to provide a multi-wavelength fluorescence detection DNA fragment detector.
The excitation light sources could be changed from LEDs to Laser Diodes (semiconductor solid-state lasers). Alternatively, they could be pulsed lasers (e.g., solid state lasers, gas lasers, dye lasers, fiber lasers). The main reason for using LEDs (i.e., Green, 524 nm) is their low cost, super brightness, and small package. Surface Mount (SMT) type LEDs could also be used, using either fiber coupled or direct butt-to-butt coupled scheme to capillaries to deliver excitation light to the separating analytes. An alternate light source for this instrument would be laser diodes in the range of 400-800 nm.
A person skilled in the art will recognize that the instrument incorporating the essence of this invention can also be used for other biomoleculer analysis. For example, by altering the separation gel or buffer, the system can also be modified to analyze biomolecules like proteins, carbohydrates, and lipids. Using a number of multi-channel cartridges of the present invention having different buffer/gel chemistries, capillaries, etc., particular buffer/gel chemistry, with matching capillary (e.g., with particular internal wall coatings and column sizes), may be easily interchanged to suit the particular sample based separation applications and run conditions, to achieve different separations, types, speeds, resolutions, etc. The same cartridge may be set aside, and later reused for conducting future separation runs. Compared to the prior art CE systems, the set up time to prepare the present CE system 200 using the cartridge 100 to run different test can be reduced significantly, since the separation column, the separation medium, and at least the detection optics requiring fine alignment with respect to the capillaries are all self contained within the cartridges. The reusability of the cartridge significantly reduces the material cost for the CE system. Also since the gel matrix with intercalated dye is hermatically sealed inside cartridge it provides a good solution for an environmentally safe/"Green" product. The fluorophore and/or gel matrix may contain carcinogens and other materials harmful to health and environment. By packaging the gel inside the cartridge, it significantly ease handling and improve safety. The cartridge may be collected and disposed of accordingly in an environmentally safe manner, or it can be recyclable, with spent parts replaced or refurbished by trained technicians to avoid harm to the environment.
With this automated and modular with integrated optics and self-aligning (non-moving micro-optical parts) multi-channel approach the operation of the instrument becomes simpler, more reliable yet provides high throughput. The cartridge 100 with self-contained, pre-aligned optics with respect to the separation channels, can be easily snapped into the CE system 200. Further, this multi-channel detection scheme could be expanded or scaled up to more than 12 or even Nth number of detection channels (e.g. 96-channels) without impairing the detection sensitivity. The other advantage of this simple time-multiplexed type detection method is that there is negligible or no cross talk between the channels compared with any other high-throughput LIF detection schemes.
While in the embodiments described above, the multiple radiation sources are at the same wavelength, it is within the scope and spirit of the present invention to configure the multiple radiation sources at different wavelengths, to complement the specific samples, sample based detection applications or gel chemistries in the different capillaries.
Incident radiation for the detection may be directed at the detection zone and/or radiation emissions from the detection zone may be output axially along the separation medium. A widened detection zone may be adopted. References are made to U.S. patent application Ser. No. 09/887,871 entitled Optical Detection in Bio-Separation Device Using Axial Radiation Input, U.S. patent application Ser. No. 09/887,953 entitled Optical Detection in Bio-Separation Device Using Axial Radiation Output, and U.S. patent application Ser. No. 09/887,872, now U.S. Pat. No. 6,529,275, entitled Optical Detection in Bio-Separation Device Using a Widened Detection Zone, all filed on Jun. 22, 2001, which are commonly assigned to BioCal Technology, Inc., the assignee of the present invention, and which are fully incorporated by reference herein.
The low cost instrument of the present invention has a disposable/recyclable multi-channel cartridge design (since, most of the cartridge body parts could be retrieved and then repackaged or reused. The only part that would be replaced are the capillaries and the gel), a fluorescence detection system, and a built-in sample handling tray (96-well plate) mechanism. Experiments have demonstrated the analyses of samples are completed in just 4 to 10 minutes per twelve-channel (twelve parallel results for twelve test samples). The DNA analyzing system is an all-in-one high throughput workstation that handles complete DNA fragment analysis from injection to detection to fragment data collection. Detection sensitivity for a single capillary using the described detection mode of the present invention is in the order of 0.02 ng of the DNA fragment in less than 10 minutes of separations (using HaeIII digest φX174 bacteriophage DNA test mix). This kind of approach for having twelve micro-channels/capillaries running in parallel produces results within 10 minutes for all twelve electrophoresed samples. This kind of separation speed and detection sensitivity is several orders of magnitude better than conventional slab gel-electrophoresis techniques.
While the invention has been particularly shown and described with reference to the preferred embodiments, it will be understood by those skilled in the art that various changes in form and detail may be made without departing from the spirit, scope, and teaching of the invention. For example, the excitation radiation source could be, for example, LEDs, Laser Diodes (semiconductor solid-state lasers), pulsed lasers (e.g., solid state lasers, gas lasers, dye lasers, fiber lasers), or other sources of radiation. LEDs (e.g., Green, 524 nm) are associated with low cost, super brightness, and small package. Alternate relative inexpensive light source for the present invention could be laser diodes in the visible, UV and/or infrared range. For example, laser diodes in the range of 400-900 nm, and more specifically in the range of 400-600 nm may be used, for example.
A person skilled in the art will recognize that the instrument incorporating the essence of this invention can also be used for biomoleculer analysis other than DNA analysis. For example, by altering the separation gel or buffer, the system can also be modified to analyze biomolecules like proteins, carbohydrates, and lipids.
By way of example and not limitation, the detection scheme of the present invention is described in connection with capillary electrophoresis and radiation induced fluorescence detection. It is understood that the present invention is also applicable to detection of analytes separated based on bio-separation phenomenon other than electrophoresis, and detection of radiation emissions other than fluorescence emissions, including other types of emissive radiation, such as phosphorescence, luminescence and chemiluminescence, as well as absorbance based detection.
Furthermore, while the separation channels in the described embodiments are defined by cylindrical columns or tubes, it is understood that the concepts of the present invention is equally applicable to separation channels defined by open channels, for example micro-channels defined by etching in a substrate (micro-fluidics type devices or bio-chips).
The transport mechanism can be configured to move the trays in a horizontal plane, and an additional transport mechanism may be provided to move the cartridge vertically to access the trays.
Accordingly, the disclosed invention is to be considered merely as illustrative and limited in scope only as specified in the appended claims.
Patent Citations
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Click Here to Watch the Whole Video in HD at GreatCanadianMale
Where Canadian Straight Boys Go Gay!
This Is Izak's Last Video Before He Heads Off to Europe. This Time I Convinced Him to Take the Plunge and Fuck Some Guy Ass. After the Shoot I Asked Him How It Was Fucking Ass and He Said "it Was Pretty Good"!
Pixels: 320X240 | Bitrate: 350K
Pixels: 640X480 | Bitrate: 700K
Izak is such a doll but we see practically nothing of him. I love the soft hair that grows on his arm above the elbow. Light, soft hair there always turns me on. Bye Izak, come back soon.


text:
In recognition of April as Child Abuse Prevention Month, Voices For Children – CASA of Brazos Valley and Scotty's House Child Advocacy Center are hosting the first ever Dog Tired of Child Abuse event:
What: Dog Tired of Child Abuse – a fun, family-friendly event that allows our community to be a voice for children who need one. Includes a hot dog eating contest, with prizes given for first, second and third place in each age category (5-8 years, 9-12 years, 13-16 years, 17 and up). Registration fee is $25 per person. The deadline to register is April 15, 2013.
When: 10 a.m. – Noon, Saturday, April 20, 2013
Where: Sue Haswell Park, 1142 E. William J. Bryan Parkway (Pavilion B), Bryan, TX
Benefit: Proceeds from the hot dog eating contest will benefit Voices For Children, Scotty's House and the abused and neglected children they serve every day.
For more information and to register for the Dog Tired of Child Abuse 1st Annual Hot Dog Eating Contest, log on to www.vfcbrazos.org.
To comment, the following rules must be followed:
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text:
PROVIDENCE, R.I.-- November 30, 1998 -- Are you an adult learner who
has always had a hankering to attend college? Like helping your community?
Here's an opportunity to do both without putting a dent in the family budget.
Through the support of Alan Shawn Feinstein, URI's Feinstein College
of Continuing Education is offering scholarships to new and returning students
in the name of his mother, Lillian Feinstein and in memory of individuals
whose lives have been tragically cut short.
In honor of the passing of Lillian Feinstein on November 15, eighteen
of these scholarships are now being offered for the coming spring semester.
Any person may apply for consideration of a scholarship. Those selected
for scholarships must agree to perform 30 hours of community service that
is required of all Feinstein scholarship recipients. New students will be
required to take a four-credit undergraduate starter course.
A committee will review the applications and award the scholarships.
The application deadline is December 18. For an application, call
Zena Fernandes at 277-5162.
For More Information: Jan Sawyer, 874-2116
Copyright © 1999
University of Rhode Island
more information about this site, contact firstname.lastname@example.com
File last updated: Saturday, February 24, 2001
The University is an affirmative action/equal opportunity
All rights reserved. URL: http://www.uri.edu/news/


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Register now for free, or sign in with any of these services:
Hey CROOKS in Washington, the hell with AMERICANS give more FOREIGN AID TO COUNTRIES THAT HATE US..........
Just left Chic Fil A at Paramus Park Mall, line 300-400 for a Chic Fil A
Just left Chic Fil A at Paramus Park Mall, line 300-400 for a Chic Fil A
Never been to a Chick Fil A as of yet, but will be there this WED. Freedom of Speach, only for special interests, think Ill buy 2 and send 1 to Barney Frank///////
Would be nice if in all this talk they would publish the salaries and benefits of "Our: BCPD Officers, just a thought
Why do Lawyers always get a bad rap, hey they went to school LOL
This a typical political "Dog and Pony" show the decesion has been made, if not why would they go out to bpnd $6.2MILLION DOLLARS to rehab 327 Ridgewood Ave Paramus, for the new "Headquarters" for the BCPD? now they are throwing the number of $47MILLION being saved by not building the BCPD Headquarters in Hackensack? Hey Kath what happened to the $8MILLION estimate a while ago, another Homeless Shelter the Taxpayers of Bergen County mean nothing.....
I dont think the voters/taxpayers of Bergen County are smart enough to vote on this issue, in todays Recors Kath is "Tooting" her horn that by spending $6+ Million to renovate 327 E Ridgewood Ave Paramus, for the new home of The BCPD, she is saving the taxpayers $47 Million instead of building a new BCPD Headquarters, I thought the Record reported that cost as $8Million these people are OUT OF CONTROL,if the decesion has not yet been made, why did the Freeholders go out to Bond last week?????????
This is a done deal, the freeholders already securing a $6 MILLION PLUS bond for the renovation of 327 Ridgewood Ave, "for the BCPD" the taxpayer means NOTHING like the $650,000.00+ Study meant.....
A Licenesed Hand Gun, here coupled with training, would have been the proper end to this story
Here again a Licensed Handgun, would have solved a lot of problems,
OMG This Park must be closed down............Just think one less Goose dumping all over the place, how sad..... has a Lawyer stepped up yet to represent the "Goose"????
About time, Go Gov.....
We need more Troopers plain and simple, we were told that the sur charge on either registration or licensse would fund more troopers, wana bet that money disappeared too>>>>>>>
Most likle a mouse ran in front of the car
Former Acting Gov. Richard "Dick" Cody shared with New Jersey Taxpayers that they were on The "Hook" for $$$$$50MILLION, concerning The Encap SCAM, does anyone know if Jersy Taxpayers are on the "HooK' for "The American Dream" and if so for how much???????
The other name for this vioctim and her "Lawyer" PARASIUTE
Gee only 60,000 illegals ( designated group) in New Jersey, dont blame "the illegals" blame the "rat" politicans that permitted it. they are so busy stealing our money they dont have time for anything else. and we love it, lets see I am not a lawyer(scum bag) I dont have a"degree" U.S. Southern Border about 1,800-2,000 miles long. U.S. Military secures Border????? P.S. was at a High School Graduation native born people talking and laughing during Pledge, three I guess "Illegals" standing at attention looking at Flag, Oh well......
Had the victim had a legal licenesed firearm he most likley would be alive, all of those involved should be in prison for life, this was no accident, they wanted to kill this man, away for life, notice how American Cities are ouit of control, letting these animals rob rape beat and kill innocent people......
What a joke, imagine if he was Italian with a garbage truck, or in the construction busisness these birds do what ever they want, rob the American Taxpayer, but Bill Clinton "Pardoned the Orange Copunty NY bunch" while they were doing time, if they voted for Hillary its all a joke Dwek for President, Bernie Madoff Sec of Treasury


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Social Security a Ponzi scheme!
I never realized it until Governor Perry showed me the light during the GOP debate but it's true: the entire Social Security system is a giant Ponzi scheme! Let's look at the evidence . . . I "invest" in Social Security and then get paid when I retire with other people's "investments". Yup. That's the ... Read More


text:
Cisco Systems has hired Barclays to find a buyer for Linksys, a unit which makes wireless routers for the consumer market, according to Bloomberg, which cites "people with knowledge of the situation."
Cisco had acquired Linksys in 2003 for $500 million.
Bloomberg contends the business could attract TV makers seeking a recognized brand and technology; the story asserts that the sale price could be a lot less than the price Cisco originally paid.
Disclosure: My wife is a Cisco employee.


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updated: 10/11/2012 1:14:24 PM
Three natural gas providers in the state are predicting winter heating bills will remain unchanged or be lower than last year. Evansville-based Vectren Corp. (NYSE: VVC), Indianapolis-based Citizens Energy Group and Merrillville-based NIPSCO all say natural gas markets should be steady and keep customers' prices stable.
October 10, 2012
News Release
INDIANAPOLIS, Ind. � Natural gas prices that for the past three years have remained stable and relatively low will again translate to manageable bills for Vectren Energy Delivery of Indiana - North (Vectren) customers. Today, the utility announced that assuming normal winter weather year-over-year, customers could see bills that are as much as 5 percent lower. Abundant gas supply in the U.S. has resulted in natural gas costs, which are passed through to customers on a dollar-for-dollar basis, remaining at levels not seen since 2001. As such, bill amounts have decreased by nearly 40 percent since the winter of 2008-2009.
�Given gas costs represent about 60 percent of winter bills, these lower prices are helping keep bill amounts down,� said Brad Ellsworth, president of Vectren Energy Delivery of Indiana - North. �As such, natural gas has continued to be the most cost-effective fuel source for home heating, especially when customers choose high efficiency furnaces. Moreover, the price of gas is expected to remain relatively stable for years to come.�
This year, based upon expected natural gas commodity costs and normal winter weather, customers can expect to pay around $515 over the five-month heating season of November through March based on the average household consumption of approximately 630 therms. Bills will vary by customer depending on the size and age of the home, number of gas appliances, number in the household, thermostat settings and levels of insulation. Keep in mind, last winter was very mild, and the average customer�s actual usage was only 510 therms. As such, bill amounts for November 2011 through March 2012 were much lower than originally forecasted.
�Despite the very positive outlook for natural gas bills, customers should still implement energy efficiency measures and find ways to use less natural gas to lower bills even further,� Ellsworth added. �Investing in high-efficiency appliances, implementing low-cost energy efficiency measures and upgrading insulation are still the right steps to undertake.�
Vectren has in place a number of tools to help customers manage their energy bills.
-Conservation Connection � Offers appliance rebates and new energy-saving tools to help customers lower their natural gas bills. All Indiana residential and small commercial natural gas customers are eligible. A conservation call center 1-866-240-8476 is also available to help customers focus on conservation. Visit Vectren.com for a list of rebates and qualifying appliances.
-Budget Bill: Under this billing plan, a customer's estimated costs for a year of gas service are spread out in equal monthly bill amounts for the year. This leveling of monthly bill amounts reduces the need to pay the full amount in the winter and spreads some of those higher bill charges into the non-heating months. Amounts are adjusted each summer for actual costs, and the customer�s credit or amount due rolls into the next Budget Bill payment for the next 12-month period. Customers can enroll for free at www.vectren.com or by calling 1-800-227-1376.
-Energy efficiency tips: Low-cost home improvement and energy conservation tips are available at Vectren.com or by calling Vectren�s Conservation Connection at 1-866-240-8476. Customers can also access how-to videos on home improvement projects such as caulking, weather-stripping or installing a low-flow showerhead at www.VectrenLiveSmart.com.
-Energy Assistance Program (EAP): State and federal utility assistance dollars are available for income-eligible customers. Those that fall within 150 percent of federal poverty guidelines should visit their local community action agency to sign up for the EAP. To apply for the EAP, customers should call 1-800-872-0371 to locate their nearest community action agency.
-Universal Service Program (USP): All eligible natural gas customers who apply and qualify for EAP will automatically be enrolled in the Universal Service Program (USP), which provides additional gas bill reductions that range from 15 to 32 percent of the total bill (not including EAP benefits) in the months of December through May. The USP, which has been in place since 2005 is the result of a collaborative effort by Vectren, Citizens Energy Group, the Indiana Office of Utility Consumer Counselor, Citizens Action Coalition and a group representing industrial customers. The program provides about $5 million in assistance each year to roughly 25,000 Vectren Indiana customers, including crisis assistance dollars, which target qualified low-income and working-poor households that need additional help to get reconnected and/or maintain heat throughout the upcoming winter.
-Share the Warmth: Those wanting to help the less fortunate with their energy bills can contribute to Share the Warmth, a nonprofit organization that assists income-eligible customers with home weatherization services. Each year, Vectren matches all public donations, up to an annual total of $200,000. Donations may be tax-deductible and can be sent to Share the Warmth, P.O. Box 931, Evansville, IN 47706-0931 or visit www.sharethewarmthinc.com.
�Regardless of lower bill amounts, we know many Hoosiers still find themselves in need of financial assistance, and we encourage them to reach out to us before they get behind on payments� said Ellsworth. �Programs like the Universal Service Program are critical in helping Hoosiers manage costs and stay warm. We encourage customers to contact their local community action agency today and apply for financial and/or home weatherization assistance.�
Natural gas is a commodity bought and sold in a national deregulated market and prices fluctuate daily due to supply and demand pressures. Vectren purchases natural gas on behalf of its customers and then passes those costs on to customers through the gas cost adjustment, which is listed on the Vectren bill as �Gas Cost Charge.� Vectren utilities are not allowed to mark up nor profit from the purchase and sale of natural gas. Only gas costs approved by the Indiana Utility Regulatory Commission are recovered from customers.
Vectren North delivers natural gas to approximately 570,000 natural gas customers in Adams, Allen, Bartholomew, Blackford, Boone, Clark, Clay, Clinton, Decatur, Delaware, Fayette, Floyd, Fountain, Grant, Greene, Hamilton, Hancock, Hendricks, Henry, Huntington, Jackson, Jay, Jefferson, Johnson, Lawrence, Madison, Marion, Martin, Miami, Monroe, Montgomery, Morgan, Orange, Owen, Parke, Putnam, Randolph, Rush, Shelby, Tippecanoe, Tipton, Vermillion, Vigo, Wabash, Warren, Wayne, Wells and White counties.
About Vectren
Vectren Corporation (NYSE: VVC) is an energy holding company headquartered in Evansville, Ind. Vectren's energy delivery subsidiaries provide gas and/or electricity to more than 1 million customers in adjoining service territories that cover nearly two-thirds of Indiana and west central Ohio. Vectren�s nonutility subsidiaries and affiliates currently offer energy-related products and services to customers throughout the U.S. These include infrastructure services, energy services, coal mining and energy marketing. To learn more about Vectren, visit www.vectren.com.
Source: Vectren Corp.
October 10, 2012
News Release
INDIANAPOLIS, Ind. - Citizens Energy Group advised today that the typical gas bill for residential customers in Marion County will decrease about $20 this winter compared to last winter, assuming normal temperatures. The typical residential customer in Marion County will likely use 629 therms of natural gas from November through March, for a total bill of about $555. This compares to about $576 the typical customer would have spent last year.
"Thanks to lower natural gas prices, we are pleased customer bills will be slightly less this winter," said Carey Lykins, President and CEO of Citizens Energy Group. "Even with the decrease in natural gas prices, we know a large number of customers will need help paying their utility bills this year. That's why we are working with community partners to direct customers to available sources of assistance."
Natural gas prices are not set by local utilities like Citizens, but rather by the market forces of supply and demand. Thanks to falling natural gas prices due to increased U.S. production, gas bills in Indianapolis are expected to decline about 24 percent since 2007. Natural gas costs are passed on to customers on a dollar for dollar basis.
Citizens expects to fund more than $1.5 million in energy assistance for low-income customers this winter. The utility is also working with elected officials and regulators to find additional financial assistance for customers, while keeping payment programs and policies as flexible as possible.
Marion County residents in need of energy assistance should call Community Action of Greater Indianapolis at 524-6966. People outside Marion County should call their local Community Action agency. Utility customers can also call Connect to Help at 211 for a variety of household assistance programs. Customers are also urged to contact Citizens, at 924-3311, if they get behind on their gas bills.
Citizens recommends customers enroll in the budget plan to help manage their utility costs. Now that most Marion County customers are enjoying the convenience of a combined gas, water and sewer bill, the budget plan can spread the costs out equally over 12 months. To be eligible for the budget plan, customers need to be current on their bill and have a good payment history. To sign up for budget billing, customers should call 924-3311.
To provide information about energy conservation and safety, Citizens will be hosting its annual Be WinterWise event this Saturday from 9 a.m. to noon outside its Customer Service Center at 2020 N. Meridian. Customers will be able to receive information about how to reduce their utility bills and free smoke and carbon monoxide alarms.
Among the energy savings tips Citizens recommends are:
-Replace your furnace filter - Changing your furnace filter once a month helps your furnace work more efficiently.
-Dial down your thermostat - A 10-degree reduction for an 8-hour period each day will reduce your heating bill 10 percent.
-Dial down the water heater - For each 10-degree reduction in water temperature, you can save up to five percent in energy costs.
About Citizens Energy Group
Citizens Energy Group is a Public Charitable Trust providing safe and reliable utility services to about 1 million customers in and around Marion County. The Public Charitable Trust means the utility is managed only for the benefit of customers and the community. Additional information is available online at www.CitizensEnergyGroup.com.
Source: Citizens Energy Group
October 10, 2012
News Release
MERRILLVILLE, Ind. � Each year, NIPSCO looks at weather predictions, market forecasts, supply trends and storage opportunities to calculate a winter bill projection to help prepare customers for the season ahead. Assuming normal winter temperatures, this year's projections indicate that bills will be among the lowest in a decade and only slightly below (0.56 percent) last winter's bills.
Over the course of the five-month winter heating season � Nov. 1 to March 31 � NIPSCO's average residential customer using a total of 624 therms could expect to pay approximately $438. This compares to $440 for a customer using the same amount of gas during last winter.
For the 11th consecutive month, NIPSCO has the lowest residential gas bills in Indiana, according to bill comparisons released in October by the Indiana Utility Regulatory Commission (IURC).
Note: Actual bills vary by customer depending on the home�s age and size, number in the household, number and age of gas appliances, thermostat settings and insulation levels.
�Market prices for natural gas continue to hold at historically low levels thanks in large part to plentiful domestic resources,� said NIPSCO CEO Jim Stanley. �This is great for our customers, especially as we head into the winter heating season, when utility bills typically reach their highest point of the year.�
While winter bill amounts are projected to be relatively flat compared to last winter, the company expects usage amounts to be slightly higher. Last winter�s usage amounts were lower than normal due to unseasonably warmer temperatures.
The cost of natural gas represents the largest portion of customer bills � on average, two-thirds of an overall bill. NIPSCO has no mark-up and makes no profit on the cost of natural gas billed to its customers. Before billing, natural gas commodity costs must be reviewed by the IURC.
While NIPSCO cannot control market conditions affecting the price of natural gas, the utility does negotiate the best possible price for the natural gas it purchases for its customers.
NIPSCO�s purchasing strategy is a multi-faceted approach that consists of buying gas in the market from a variety of supply sources at different times throughout the year. The ability to utilize gas storage helps to offset market price volatility.
NIPSCO�s gas distribution system consists of three on-system storage facilities and connects to seven interstate pipelines providing access to eight major North American supply basins.
Billing and Payment Options
Customers who are experiencing financial difficulties are encouraged to call NIPSCO�s 24-hour Customer Care Center as soon as possible to determine what options might be available to offer help. Some of those solutions include:
-Payment Assistance Programs: Based on income levels, customers may qualify to receive state and federal utility assistance dollars as well as support funds from separate NIPSCO programs by visiting their local community action agency.
-BudgetPlan: A free service to all NIPSCO customers to help manage their monthly energy bills by spreading out gas costs over an entire year.
-NIPSCO Choice Program: Gives customers an opportunity to choose an alternative natural gas supplier for their home or business. Customers can purchase gas from an alternative supplier, but NIPSCO is responsible for maintaining the pipeline that delivers the natural gas.
-DependaBill and Price Protection Service (PPS): DependaBill provides a fixed monthly bill with no annual reconciliation, while the PPS option allows customers to fix or cap their per-unit cost of natural gas.
For more information on billing options and payment assistance, visit NIPSCO.com/BillingPayment.
Energy and Money Saving Programs
Through NIPSCO�s energy efficiency programs, customers can pinpoint ways to manage their energy usage, and in turn their bills. A full list of programs available to NIPSCO customers to help manage energy use can be found at NIPSCO.com/SaveEnergy.
NIPSCO, with headquarters in Merrillville, Ind., is one of the seven energy distribution companies of NiSource Inc. (NYSE: NI). With more than 786,000 natural gas customers and 457,000 electric customers across the northern third of Indiana, NIPSCO is the largest natural gas distribution company, and the second largest electric distribution company, in the state. NiSource distribution companies serve 3.8 million natural gas and electric customers primarily in seven states. More information about NIPSCO is available at www.nipsco.com.
Source: NIPSCO


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- Accounting Software Integration
- Apple Pay
- Batch Credit Card Processing
- Credit Card Machines
- eCheck (ACH) Payments
- Electronic Billing & Invoicing
- EMV Credit Card Processing
- Fraud Management Tools
- Interactive Voice Response
- Merchant Accounts
- Merchant Lending Program
- Mobile Processing
- Payment Gateway
- PayPal
- PCI Compliance
- Point-to-Point Encryption (P2PE)
- POS Systems
- QuickBooks™ Integration
- Hosted Payment Form
- Tablet Point of Sale
- Tokenization
- Virtual Terminal
A Sampling of Satisfied Customers:
iPad Point of Sale System: Retail Goes Mobile
Imagine one of your sales associates comes across a customer in a quandary. The customer was looking for an item to buy, and had even found it in an ad or online. But when seen in person, the item just wasn't right or didn't live up to expectations.
Instead of losing a sale, your sales person instead can whip out an iPad filled with all of your store's merchandise – including pictures, descriptions and prices of each and every item – and help the customer find something else to purchase.
The customer can buy the item – and pay for it – right there or have it shipped to his or her home.
BluePay's iPad Retail POS System: Show and Sell
- An interactive point-of-sale retail system
- Provides customized selling to clients
- Wireless payments
- Inventory maintenance
- Retail workflows
- Track gift card sales
- Look up inventory
- Accept on-screen signatures
- Print or e-mail receipts
- And more
Cross sell other items. Upsell to better items. All is possible when your sales people show your additional inventory as they speak to customers.
Our iPad POS System Is Like Having an Additional Salesperson on the Floor….
…One with Encyclopedic Knowledge of Your Inventory!
BluePay's iPad POS System: Retail Never Had it So Good
- The Apple iOS application is highly mobile (delivered via iPads) and lets your salespeople work – and make sales – from anywhere in your store.
- Allows you to get customer payment from anywhere in your business. In the aisle. At the counter. By the dressing room or closeout bin.
- Keeps all SKUs and other information about your merchandise in one place. Your sales people can even access inventory in your firm's warehouse or in other stores (if applicable).
- Add new merchandise to your offerings list and it will be uploaded to all of your devices at the same time.
- Want to know what sold last week, last September, last night? Our iPad retail POS solution lets you access all sales and orders that took place at any time you designate. Reports generation becomes exceptionally easy.
- Manage your sales reports by sorting them for date, price, and even by salesperson. Find out what's moving and what's not. See which sales team members are outselling others.
- Transfers all sales data into a spreadsheet. Move that spreadsheet into your store's bookkeeping system for quick and easy reconciliation.
BluePay's iPad Point of Sale System Features and Benefits
- Places your top sellers and favorite items where you can access them quickly for viewing.
- Our "My Favorites" tab allows you – or your salespeople – to keep your company's top/favorite/best items right at your sales team's fingertips.
- Cash is still king.
- Customers don't have to pay with a credit/debit card; they can still pay with cash. Or with a check. They can even pay for their purchase at your cash register. All are synced to your iPad POS retail system.
- Add new merchandise/SKUs at any time from any device.
- All new items will upload to all devices at the same time.
- Swipe customer credit cards – or input the information manually.
- You also can capture digital signatures from anywhere in the store.
- Manage your staff.
- See who's selling the most. When are they selling it? Are they upselling? How many returns do they receive?
- Update your sales staff members' profiles.
- Change at will who can and cannot access the iPad retail POS system.
- Are your customers from north of the border?
- Our point-of-sale iPad system for retailers allows you to toggle back and forth between U.S. and Canadian currencies, giving your customers the ability to pay with Canadian dollars.
- Print customer receipts wirelessly.
- Or e-mail customers their receipts.
- Works easily with your other POS hardware.
- BluePay's mobile POS system works seamlessly with your store's other terminals, cash registers and printers.


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Kymco MXU 300 2013
2013 Kymco MXU 300 specifications, pictures, reviews and rating
Picture credits - Kymco. Click to submit more pictures.
|Kymco's profilation of this bike
|Designed primarily for the utility-minded construction, farm, hunting, fishing and ranch markets, the new KYMCO MXU 300 follows in the footsteps of the popular MXU 250. Plus - it boasts a more powerful 270cc, 4-stroke, liquid cooled, SOHC engine with rock-solid shaft drive.
|General information
|Kymco MXU 300
|Do you know this bike?
Click here to rate it. We miss 2 votes to show the rating.
|Price as new (MSRP):
|US$ 3899. Prices depend on country, taxes, accessories, etc.
|Engine and transmission
|270.00 ccm (16.48 cubic inches)
|Engine type:
|Single cylinder, four-stroke
|18.30 HP (13.4 kW)) @ 7000 RPM
|Fuel system:
|Fuel control:
|Electric/Pull back up
|Lubrication system:
|Wet sump
|Cooling system:
|Transmission type,
final drive:
|Shaft drive (cardan)
|Automatic CVT w/forward, neutral and reverse
|Chassis, suspension, brakes and wheels
|Front suspension:
|Dual A-arm
|Rear suspension:
|Swing arm
|Front tyre:
|Rear tyre:
|Front brakes:
|Single disc
|Rear brakes:
|Single disc
|Physical measures and capacities
|Dry weight:
|216.8 kg (478.0 pounds)
|Power/weight ratio:
|0.0844 HP/kg
|Seat height:
|808 mm (31.8 inches) If adjustable, lowest setting.
|Overall height:
|1,110 mm (43.7 inches)
|Overall length:
|1,808 mm (71.2 inches)
|Overall width:
|1,049 mm (41.3 inches)
|Ground clearance:
|210 mm (8.3 inches)
|1,184 mm (46.6 inches)
|Fuel capacity:
|12.49 litres (3.30 gallons)
|Other specifications
|Carrying capacity:
|Heavy-Duty Front and Rear Equipment Racks
|Factory warranty:
|2 Year Warranty
|Color options:
|Green, Red, Black
|C.A.R.B. Certified with USFS-Approved Spark Arrestor.
|Further information
|Parts and accessories
|Check out parts and accessories from our other partners.
|Ask questions
|Join the 13 Kymco MXU 300 discussion group.
|Insurance, loans, tests
|Check out approximate insurance costs here. Search the web for dealers, loan costs, tests, customizing, etc.
|Related bikes
|List related bikes for comparison of specs
Rate motorcycles you knowRate motorcycles you know and help your fellow riders to find the bike that is right for them. Bikez has the most users and the most extensive motorcycle rating on the net. It includes reliability, design, engine performance, accident risk, repair cost, offroad capabilities, racing track capabilities and touring capabilities.
The ratings are also used to rank the 100 most popular MC models each year since 1990. The lists are adjusted weekly as new ratings are added. You can check out the rating of the 2013 Kymco MXU 300 and compare it to other bikes here.
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The culmination of any good outdoor party has to be roasting marshmallows around a bonfire. A slightly charred, gooey marshmallow sandwiched between chocolate and graham crackers is a timeless combination that will remind you of your childhood. This s'mores brownie recipe is based on all of those classic flavors, but is a much more manageable version, perfect for parties, picnics, or off-season cravings—no fire pit required!–Karen Covey
LC Indoor, Not Outdoor, Brownies Note
These s'mores brownies work swell around a campfire, but rest assured, no outdoor pyrotechnics are required.
S'Mores Brownies Recipe
- Quick Glance
- Makes 12 to 16
- For the crust
- 6 tablespoons (3 ounces) unsalted butter, plus more for the pan
- 1 1/3 cups graham cracker crumbs, finely ground (from about 12 big rectangles)
- 2 tablespoons granulated sugar
- Pinch kosher salt
- For the brownies
- 12 tablespoons (6 ounces) unsalted butter, cut into pieces, plus more for the pan
- 6 ounces semisweet chocolate, roughly chopped
- 1 1/4 cups plus 1 tablespoon granulated sugar
- 3 large eggs, at room temperature
- 1 1/2 teaspoons vanilla extract
- 3/4 cup all-purpose flour
- 1/2 teaspoon kosher salt
- 12 to 16 large marshmallows
- Make the crust
- 1. Preheat the oven to 350°F (177°C).
- 2. Butter the bottom and sides of an 8-by-8-inch baking pan and line it with parchment paper cut to fit the bottom of the pan. Coat the parchment paper with butter.
- 3. In a small saucepan over low heat or in a bowl in the microwave, melt the butter. In a medium bowl, combine the graham cracker crumbs, sugar, and salt. Add the melted butter and mix well. Press the crumb mixture evenly over the bottom of the prepared pan with a spatula and bake just until the crumbs are golden brown around the edges, about 10 minutes.
- Make the brownies
- 4. Place a medium heatproof bowl over a saucepan filled with 1 to 2 inches of water (make sure the water doesn't touch the bottom of the bowl). Add the butter and chocolate to the bowl and place the saucepan over very low heat, stirring occasionally, until the chocolate melts. Remove the pan from the heat and let cool for 5 minutes.
- 5. Add the sugar to the melted chocolate mixture and stir to combine. Slowly whisk in the eggs, 1 at a time, and mix until incorporated. Add the vanilla and stir until incorporated. Slowly add the flour in 2 batches and stir just until incorporated after each addition. Stir in the salt just until combined. Do not overmix.
- 6. Scrape the batter onto the graham cracker crust and bake until the brownies are set and the center is still a bit soft, 30 to 40 minutes. They should be soft but still a bit gooey when tested with a toothpick—you want moist crumbles on the toothpick. Start checking them after 25 minutes. Do not overbake the brownies, but don't underbake them, either. Remove the pan from the oven, adjust the oven rack to the top position, and turn on the broiler.
- 7. Place the marshmallows evenly over the brownies and slide the pan back in the oven. Broil until the marshmallows are softened and lightly toasted, 45 seconds to 1 minute. (Do not turn your back on the marshmallow-topped brownies.) Carefully pull the pan from the oven and, using the back of a spoon, gently press down on the marshmallows to flatten them slightly. (You want to do this slowly so that they don't stick to your spoon.) Return the pan to the oven and broil until the marshmallows are nicely toasted, another 15 to 30 seconds. Remove the pan and allow the brownies to cool for at least 1 hour. [Editor's Note: We know, we know, this is nearly impossible. But it allows the brownies—in all their magnificent layers—to fully set. Trust us.]
- 8. Run a sharp knife under warm water and then run the knife along the edges of the pan. Cut the s'mores brownies into squares, ensuring each square has some ooey, gooey, toasted marshmallow on top. After each cut, run the knife back under warm water, wipe clean, and repeat.
Hungry for more? Chow down on these:
- Brownies in a Cast Iron Skillet from Amanda's Cookin'
- Campfire Bars from Cookies and Cups
- S'mores Brownies from Leite's Culinaria
- Gianduia "Nutella" Brownies from Leite's Culinaria
S'Mores Brownies Recipe © 2013 Karen Covey. Photo © 2013 Cassandra Birocco. All rights reserved.