Modulus Single Tube Fluorometer

TBS-380 Mini-Fluorometer

Picofluor Fluorometer

Modulus Single Tube Fluorometer
Richard Lasker, Director
Brabant Research, Inc.
Bent Mountain, Virginia
www.brabantresearch.com

 
Our work centers about analyzing the nutrient content of raw fruits & vegetables. This helps us determine what is, and is not, in the food we all consume for our nutritional needs. The accumulation of this data, and the comparison of the USDA "label" nutritional levels that are used throughout the food industry, blanket without any confirmation, will help us design better growing programs coupled with better varieties to improve human nutrition and therefore decrease human illness issues. This research will be done through Extraction of vitamins, amino acids and other nutritional compounds via wet chemistry to quantify the levels of nutritional compounds in raw fruits & vegetables. We entirely self-fund the work we do in human nutrition. We have, to this date, received no monies from any source. Every dollar we make on contracts, royalties, co-operative projects or consulting we spend doing the nutritional research. This includes growing varieties in order to be able to determine how much of an increase we can attain in the selected fruits & vegetables in order to qualify selections and delineation. Every dollar we do not have to spend on necessary equipment would help speed the end-result: identifying those foods that can help prevent/cure diseases. Our work is unique in that no one, anywhere, has absolutely no agenda other than the identification and improvement of the foods consumed by us. All groups we have met all have their own agenda which often conflicts with the data and results of their work. No one is doing complete nutritional analysis of commercially grown produce. Even cancer and medical research trials utilize the USDA/FDA "label" nutritional values in their calculations. Hence why
the whole foods trials are always unsuccessful. Our preliminary work, in bone cancer and lymphoma has a 100% success rate preventingrelapse after initial chemo/radiation therapy.

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Modulus Single Tube Fluorometer
Sujey  Carro, Thesis Student (graduate-research)
UMET School of Environmental Affairs
San Juan, Puerto Rico
www.umet.edu

My project is to determine if:
1) Apoptosis is the mode of cell death using the mitochondrial transmembrane potential as an indicator.
2) ROS generation, ATP determination to see if there is mitochondrial dysfunction, impairment of cellular respiration and if the generated ROS are contributors of cellular oxidative stress.
3) Adduct formation in mitochondrial DNA. Since we already know there is adduct formation with Calf thymus DNA and nuclear DNA we would expect to see adducts in the mitochondrial DNA.

All of this will be assessed after exposing cancer cell line A431 to three different concentrations at three different times of exposure to potential anticancer drug NBQ.

I plan to publish my results to gain experience in the professional scientific community.

1)  For apoptosis we would we are currently using mitochondrial membrane permeabilization as an indicator of apoptosis.
2)   Our NBQ drug agents are autoflourescent and we propose to measure how much the adherent A431 cells uptake the drug. We can estimate the amount of drug uptake by cells measuring the absorbance change in the RPMI residual medium that will be removed every 24 hours over a 72 hour exposure period. Subtracting the amount of absorbance of the initial drug dose with RPMI from the RPMI residual will result in the change in absorbance of drug and RPMI over the 72 hours. The absorbance readings can be made with a UV-Spectrophotometer.
3)   For ROS we would like to measure hydrogen peroxide production measuring fluorescence with horseradish peroxidase.
4)   We will determine ATP using luciferase luciferin bioluminescent kit.
5)   Adduct detection would be performed by isolating mitochondria, isolating mtDNA, digest mtDNA and then use the HPLC-ESI MS/MS for detection.

As part of a Risk Management we are trying to discover the mechanism of action of these synthesized potential anticancer drugs. We are using the cancer cell line A431 in order to complete the pre-clinical phase of this study (in vitro). This equipment is an essential tool that will help us to discover the mechanism of action of the drug using the mitochondrion as a target organelle. My budget is small but I am much exited about completing it to acquire my Masters Degree.

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Modulus Single Tube Fluorometer
Chin-Chuan Wei, Assistant Professor
Southern Illinois University Edwardsville
Chemistry Department
Edwardsville, Illinois
http://www.siue.edu/~cwei

The main focus of Dr. Wei's laboratory is to study the mechanism of reactive oxygen species (ROS) generated from non-phagocytic NADPH oxidases (NOXs) that play important roles in cancer and disease development. Using the recombinant DNA/protein technology, we express and/or purify the desired proteins fused with fluorescent proteins or purification tags for structural and functional studies.
List of Assays = nucleic acid and protein quantitation , gene expression.
We request a Modulus Fluorometer that allows us to perform routine molecular biology techniques, such as nucleic acid and protein quantitation as well as gene expression. We intend to use this instrument for the measurement of ROS levels that are related to the function of NOXs both in vitro and in vivo in order to understand their molecular mechanism.

The instrument will facilitate our research on the mechanism of ROS generated from NOXs at Southern Illinois University Edwardsville (SIUE), a primarily undergraduate institute, and it will be also used in biochemistry laboratory and advance biochemistry course for gene expression experiments using green and red fluorescent proteins.

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Modulus Single Tube Fluorometer
Betsy Martinez-Vaz, Assistant Professor of Biology
Hamline University
Biology Department
Saint Paul, Minnesota
http://www.hamline.edu/cla/acad/

The primary focus of my research is to investigate the effects of sub-inhibitory concentration of antibiotics in bacterial gene expression and metabolism. My laboratory uses Escherichia coli K12 strain MG1655, a model laboratory organism, to study how gene regulation is affected by sub-inhibitory concentrations of antibiotics. Most research on antimicrobial drugs has focused on microorganisms isolated due to their high level of resistance to one or to multiple antibiotics. Bacteria are frequently exposed to low concentrations of antibiotics in laboratory environments and in their natural surroundings; the cellular effects of sub-inhibitory concentrations of antibiotics in bacteria have not been studied in detail. The significance of this research is twofold. First, elucidating how bacteria respond to sub-inhibitory concentrations of antibiotics is essential to understand the ways in which microbes overcome the effects of antimicrobial drugs. Secondly, understanding the transcriptional responses triggered by antibiotics is crucial to elucidate the natural function of antibiotics. Antibiotics were originally discovered because of their ability to adversely affect microbial species. However, many antibiotics are naturally occurring molecules (produced by microbes) which cellular function is not fully understood. My research laboratory is located at Hamline University, a primarily undergraduate institution in Saint Paul, Minnesota. Accordingly, my research program focuses on using the effects and the modes of action of antibiotics as a framework to introduce undergraduate students to scientific research while teaching the principles of molecular biology and microbiology. In order to study how antibiotics affect gene expression and trigger cell-signaling responses, my students are in the process of constructing a collection of GFP transcriptional fusions, which includes the promoters for the main transcriptional regulators present in Escherichia coli. The next step of their research will consist of measuring the activity of the GFP fusions in the presence of different classes of antibiotics at several concentrations.
By identifying E.coli promoters that are responsive to antibiotics, we will get insights into the following questions:
1) Does exposure to low concentration of antibiotics triggers specific gene expression patterns in the cell?
2) Can antibiotics act as signaling molecules?
This will allow students to learn about the role of antibiotics and the regulatory networks that control antimicrobial resistance in microbial cells.
List of Assays = 1) Measurement and quatification of fluorescence from GFP-transcriptional fusions.
2) Measurement and quantification of nucleic acids
3) measurement of microbial growth by monitoring absorbance at 600 nm.
The main application of the Modulus laboratory Fluorometer will be to quantify gene expression by measuring the activity of GFP-promoter fusions. In addition, the fluorometer will be useful to quantify nucleic acids prior to PCR and other molecular biology experiments.

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Modulus Single Tube Fluorometer
Timothy Mattes, Assistant Professor
The University of Iowa
Civil and Environmental Engineering Department
Iowa City, Iowa
www.cee.engineering.uiowa.edu

In general, I am interested in the mechanisms that govern evolution of xenobiotic biodegradation pathways.  I am also interested developing molecular tools for the detection and quantification of aerobic bacteria that degrade vinyl chloride, a known
human carcinogen, in the environment.
List of Assays = Molecular biology techniques such as cloning, sequencing, real-time PCR, real-time reverse transcription PCR, in vitro transcription
We are starting to develop techniques for real-time PCR and real-time RT-PCR of specific catabolic genes using an internal mRNA reference technique for quantification.  This requires that we accurately quantify DNA and RNA using fluorescence techniques (e.g. Picogreen, ribogreen)

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TBS-380
Nick Quirke, Professor
Imperial College
Chemistry Department
London
www.ch.ic.ac.uk/quirke

Theoretical and experimental investigation of nanoscale fluid flow.  Please see website for extended description. Initially measurement of the transport of fluorescein-labelled nanoparticles through carbon nanopipes. The development of diagnistic medical devices and also drug delivery applications. The instrument will allow us to greatly speed the gathering of data by providing a dedicated resource to the suite of nanodiffusion experiments we are conducting.

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TBS-380
Craig Simmons, Assistant Professor
University of Toronto
Institute of Biomaterials and Biomedical Engineering
Toronto, Ontario Canada
www.ibbme.utoronto.ca

We are a group of three new principal investigators (Radisic, You, and Simmons) in the Institute of Biomaterials and Biomedical Engineering at the University of Toronto. Our common research interest is tissue engineering. In each of our labs, we are combining bioengineering, biomaterials, microdevices, and cell and molecular biology with the goal of investigating and developing new ways to regenerate tissue. The specific goals of our three labs are:
Dr. Radisic’s Laboratory for Functional Tissue Engineering: To engineer functional myocardial tissue by developing novel tissue engineering systems. Focus is on the development of novel biomaterials systems that deliver multiple cell types and of bioreactors that provide appropriate biophysical stimuli (electrical, mechanical, oxygen tension) to improve the functionality of regenerated heart muscle.
Dr. You’s Laboratory: To discover new strategies for bone regeneration by studying the fundamental mechanisms by which osteocytes respond to mechanical forces and regulate osteogenesis. Microfabricated systems are used to study these mechanisms in vitro.
Dr. Simmons’ Cellular Mechanobiology Laboratory: To determine how mechanical forces regulate cell function, and to use this knowledge to develop improved therapies to treat or replace diseased tissues. Specific foci are heart valve disease and regeneration, and stem cell-based skeletal tissue regeneration.

List of Assays = Hoechst 33258, RiboGreen, PicoGreen

We have at least three immediate applications for the TBS-380:
1) As post-doctoral fellows, we each used the Hoechst 33258 assay to quantify DNA content in tissue engineering scaffolds. As new independent investigators, we intend to use this assay for the same application in our own labs;
2) The Simmons lab does transcriptional profiling of small numbers of cells isolated from heart valve tissue by laser capture microdissection (microgenomics). Because of the limited sample, we use RiboGreen to quantify the amount of RNA obtained prior to RNA amplification and transcriptional profiling by RT-PCR or microarray;
3) All three of our labs use microfabricated devices and MEMS to study cell biology. The small size of these devices limits the number of cells that are available for analysis. We intend to use the PicoGreen assay as an indirect method to quantify the number of cells in our microscale devices.

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TBS-380
Wanda Reygaert, Assistant Professor
Oakland University
School of Health Sciences
Rochester, Michigan
http://www2.oakland.edu/shs/mls/

I am finishing some previous  research so that the results can be written up and submitted for publication.
List of Assays = 4-MU
Studying transcription regulation in E. coli.  Without the grant I cannot afford to do fluorescent assays, which are a huge part of my research. I have very little in start up funding for my research. Without  enough data I can't even apply for external funding. This instrument would enable me to finish my research and get published.

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TBS-380
Aparna Palmer, Associate Professor
Mesa State College
Biological Sciences Department
www.mesastate.edu

The Biological Sciences Department at Mesa State College is committed to offering our students the highest quality of undergraduate education.  With that goal in mind, the research that is conducted by our faculty members is always done in collaboration with our students.  The Molecular Genetics Research Laboratory at Mesa State provides the space and equipment for a diversity of studies including those on the population genetics of mistletoe (Dr. Kristy Duran), the molecular basis for cell motility (Dr. Kyle McQuade), the molecular phylogenetics of polychaetous worms (Dr. Aparna Palmer), the molecular phylogenetics of rodents (Dr. Aparna Palmer), the population genetics of tropical plants (Dr. Aparna Palmer), the molecular phylogenetics of snakes (Dr. Steve Werman), the population genetics of salamanders (Dr. Steve Werman) and the characterization of cave microbes (Dr. Denise McKenney).  Each of these projects allows faculty members to advance their research goals while training undergraduates in techniques such as DNA/RNA extraction, DNA amplification, DNA/RNA purification, cycle sequencing, and gel electrophoresis.  Students also learn how to use the automated sequencer and how to perform data analysis.  Our undergraduates also learn many of these skills in formal classroom settings; courses such as Genetics, Molecular Genetics, Forensic Molecular Biology, Microbiology, Cellular Biology, Immunology, and Attributes of Living Systems rely upon the equipment in the Molecular Genetics Laboratory for hands-on training of students.  In sum, the research goals of the Biological Sciences Department at Mesa State are intertwined with our focus on offering a superb undergraduate education; as each faculty member gains a foothold on his/her research area, our students learn important skills that will allow them to compete effectively for jobs and positions in graduate school. We are primarily interested in using the PicoGreen and/or the Hoechst 33258 assays; however, we are also considering the use of the RiboGreen assay for the future.
The main application in which we are interested is DNA quantification/quantitation.  In the future, we will also need to use the instrument for RNA quantification/quantitation.  The TBS-380 fluorometer would greatly benefit our research because it would allow us to determine the concentration of our DNA samples accurately.  This is a critical issue when working with samples that may originate from such diverse sources as microscopic worms, cave bacteria, salamander tail clips, and leaves.  Each DNA extraction is unique in its yield because of the nature of tissue sample, the type of extraction procedure used, and the skill of the investigator—thus, each DNA sample needs to be quantified.   Accurate determination of DNA concentration is the key to successful PCR and cycle sequencing.  Currently, faculty members and students “eyeball” the relative concentration of each sample based on gel electrophoresis. Depending upon the experience of the investigator, this may or may not yield successful results when it comes to using that DNA for PCR or cycle sequencing.  Having a fluorometer in the research lab would allow for quantitation to be more uniform and reliable, especially in the case of minute quantities of DNA.  This, in turn, would increase our effectiveness in using the DNA samples for other procedures.  The same benefits would be seen when it comes to formal classroom activities that involve the use of DNA samples.  A fluorometer would allow learning to become more effective and progress to occur at a higher rate, both in the research lab and in the classroom.

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TBS-380
Antonio Izzo, Assistant Professor
Elon University
Biology Department
Elon, North Carolina
http://facstaff.elon.edu/aizzo/

My research involves a wide range of basic and applied questions focused on soil and root-associated fungi. Most recently I have been involved with collaborative research (with Mark Mazzola at the USDA-ARS) examining how the changes in fungal community composition relates to plant health, and how we might best be able to manage these fungi to promote plant health. As the diversity of fungi in the environment is very high, and many are resiliant to culture-based study, I use molecular approaches (DNA) to track community composition and diversity.
List of Assays = DNA Quantitation using Hoechst 33258 Dye
During my recent post-doctoral work with the USDA I developed a macroarray approach based on reverse dot blot analysis that allows the tracking and rapid identification of fungi in complex environmental samples. With this technique, PCR product from known samples is used to construct the array while PCR product from environmental samples (e.g. soil, roots, etc.) is used as the probe. Two aspects of DNA quantitation are critical for proper use of this approach. First, PCR products that are spotted on membranes to construct the array must be in equal concentrations. Second, PCR product from different environmental samples that are to be compared must be at equal concentration. Having access to accurate DNA quantitation will allow me to utilize the macroarray approach to its highest power, and give me practical means to develop customized arrays for a wide range of microbial ecology questions. I have worked with this instrument in my previous position and know that it is reliable and accurate at the levels necessary for these projects.

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TBS-380
Dr. Christian Happi, Malaria Research Laboratories
Institute for Advance Medical Research and Training
University of Ibadan
Ibadan

My research over the years has focus in understanding the in vitro and molecular basis of resistance of Plasmodium falciparum (the agent of malaria) to antimalarials and developing new molecules against the malaria parasites in Nigeria and Africa. Thus, my research depends primarily on screening the susceptibility of malaria parasites to antimalarials in vitro.  However, basic microscopic technology used for quantification of parasites for in vitro screening for drug development for malaria is tedious, time consuming and often subjective. The development and use of radioisotopes for in vitro screening of antimalarial drugs provided a more sensitive alternative to microscopy.  Unfortunately this technique is less attractive because of practical problems in malaria endemic regions where access to radioisotopes is limited or hindered and disposal constitutes a major public health hazard and policy problems.  Therefore, fluorescence-based assays are alternative ways to perform antimalarial drugs screening without radioactive usage.  We intent to use TBS-380 as an alternative to a 96-well plate reader in order to adapt the SYBR green assay to determine the susceptibility of patients isolates of Plasmodium falciparum to antimalarial drugs in Nigeria, Africa.   This will enable us provide early warning signals for the emergence and spread of parasites resistant to new antimalarial drugs, especially artemisinin derivatives and partner drugs currently used for treatment of acute uncomplicated malaria in Nigeria and many other Africa countries.
List of Assays = SYBR Green assay
Microscopic assay based on the malaria parasites morphology after exposure to antimalarial drugs.We intent to use TBS-380 as an alternative to a 96-well plate reader in order to adapt the SYBR green assay to determine the susceptibility of patients isolates of Plasmodium falciparum to antimalarial drugs in Nigeria, Africa.   This will enable us provide early warning signals for the emergence and spread of parasites resistant to new antimalarial drugs, especially artemisinin derivatives and partner drugs currently used for treatment of acute uncomplicated malaria in Nigeria and many other Africa countries. TBS-380 provides us a unique opportunity to adapt in collaboration with Dr. Martin Smilstein at the Portland VA Medical Center, the fluorescence based SYBR green assay as an alternative way to perform antimalarial drugs screening without microscope and radioactive usage (a major public hazard in African countries because of lack facilities for adequate disposal).

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TBS-380
Dr. Sabine Rech, San Jose State University. Using the fluorometer for DNA and protein quantification. First working on the isolation of halogenation enzymes from acorn worm associated bacteria. The acorn worm is an invertebrate marine worm which lives in burrows beneath beaches. Second working on elucidating the microbial diversity in salt marsh soil under restoration using restriction fragment length polymorphism.

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TBS-380
Dr. Martin Smilkstein, Portland VA Medical Center. The primary focus of the Riscoe lab is antimalarial drug discovery, including ongoing synthesis and testing of numerous novel compounds, and investigation of novel combinations of existing drugs. The lab has developed and published a simple and inexpensive fluorescence-based assay. The drug-development research capacity of nations most afflicted with malaria and the ability to monitor drug resistance in the clinical setting could be transformed by the availability affordable methods. For this reason, Dr. Smilkstein has done preliminary work showing that the TBS-380 can serve as an alternative to a 96-well plate reader in the laboratory setting using culture-adapted parasites, and plans to investigate whether the same method will succeed in on-site drug resistance testing of clinical isolates.

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TBS-380
Prof. Bruce Benz, Texas Wesleyan Univeristy. All four faculty supervise undergraduate student research with collective research goals including: Quantification of DNA from archaeological and forensic samples; Quantitate RNA for use in microarray analyses for gene expression in Drosophila; Using molecular methods to characterize the carbon dioxide mutation (DLY) in Drosophila; and quantification of proteins in signal transduction and cell adhesion in immune cells.

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TBS-380
Assistant Prof. Matthew Lorincz, University of British Columbia. By “targeting” specific HMEs to an integrated transgene, I propose to directly test whether specific histone modifications protect against or promote targeting of the de novo DNA MTases in cis. The fluorometer will be used for quantifying DNA in chromatin immunoprecipitation experiments as well as general laboratory DNA preparations. Accurate measurement of DNA in ChIP experiemnts is critical for subsequent real-time PCR analyses.

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Picofluor Fluorometer
Dr. Janet Bayleran, Eastern Maine Medical Center, Molecular Research Laboratory. PicoGreen dsDNA Quantitation Assay for Array CGH analysis of breast cancer patients. Goal is to identify genomic profiles associated with an increased risk of recurrence, thus improving prognosis and therapeutic intervention.

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Picofluor Fluorometer
Noorjehan Joonus at the Central Health Laboratory at the University of Mauritius has been granted a Picofluor Handheld Fluorometer. She is working in collaboration with Netria Laboratories in the UK to develop a fluorescence quenching method for the measurement of glycated protein and glycated hemoglobin in blood. This method will provide an inexpensive test for monitoring glycemic control in diabetics.

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Picofluor Fluorometer
Brian Rude, is an Associate Professor of Animal & Dairy Sciences at Mississippi State. He was granted a Picofluor Handheld Fluorometer to use for research on the diets of free ranging animals to determine the amount of forage being consumed. Fluorescent microspheres will be used with the forage species to determine forage intake. This is done by calculating differences in forage microspheres and fecal microspheres. This simple and cost-effective technique would revolutionize nutritional research and management of grazing animals.

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Picofluor Fluorometer
Professor Susan Jackels of Seattle University's Chemistry Department was granted a Picofluor Fluorometer for research into developing a simple, affordable and appropriate Ochratoxin A test that can be applied to green coffee samples at coffee processing sites in Nicaragua and other Central American countries. The facilities in Nicaragua and other countries may not have electricity or lab space, so the battery operated Picofluor is ideally suited to their needs.

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Picofluor Fluorometer
Dr. Jiang Zeng of the University of Toronto at Mississauga received a Picofluor Handheld Dual Channel Fluorometer for his work with sensor developments involving oligonucleotides or genomic DNA.

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Picofluor Fluorometer
Tina Davis of South Cobb High School's Academy of Mathematics and Medical Sciences has been awarded a Picofluor Handheld Fluorometer for assaying protease activity using Molecular Probe's EnzCheck Assay. Advanced students in their magnet program will be using this fluorometer.

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TD-700 Grantee - Obsolete Instrument
Cornell University
University of Texas Southwestern Medical Center
University of Colorado at Boulder
Murray State University
University of Missouri

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