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The
Biomolecular Research Facility has a Molecular Devices SPECTRAmax
Gemini EM fluorescent plate reader. The instrument is located in
Jordan Hall room 1074, in the Biomolecular Research Facility.
Users need to contact Dr. John Shannon of the Biomolecular Research
Facility at 434.243.9399 or by email at jds1c@virginia.edu to establish an
account on the computer which controls the instrument. Dr. Shannon will
offer basic training for the operation of the instrument. Instructions to get
started are available.
Some applications for the instrument are:
- measuring live and dead cells in the same well, using Promega
MultiTox reagents to measure proteinases specific for live and dead
cells. This assay takes advantage of the instrument's ability to
measure two fluorophores in the same assay and to scan wells to look
for clumps of cells.
- measuring glycogen content of cultured cells
- quantitation of DNA and RNA with PicoGreen or other dyes
- cathepsin assay
- measuring relative cell numbers with Alamar Blue
- intracellular calcium
- sensitive protein estimation
- cholesterol assay
One user is quantitating DNA at 5 ng/mL.
Features of the SPECTRAmax instrument are:
- dual monochromators (bandwidth 9 nm) allowing any fluorophore to be
read within the range of 250 to 850 nm
- fluorescent spectra capability
- top or bottom reading of plates for adherent cells
- well scanning
- kinetic measurements
- 384, 96, 48, 24, 12, 6 well plates
- heating of plates
- shaking of plate
- SoftMax Pro 4.7 software for data analysis.
- claimed sensitivity of 3 fmol/well of FITC with top
reading
This instrument does not measure absorbance.
It can perform some time resolved fluorescence measurements but not
polarized fluorescence.
The instrument can be used as a luminometer for some applications.
It does not have the sensitivity of a dedicated luminometer, nor does
it have injectors required for flash luminescence assays e.g. Promega
Dual Glo Luciferase assays.
Detection limit: 10 amol/well alkaline phosphatase 200 μL/well
(obtained with Emerald IITM reagent from Applied
Biosystems).
Users supply their own plates. For top read fluorescence, all black
plates are recommended. For luminescence, white plates which should be
kept out of light to reduce light emission from the plastic.
The instrument is user operated. The current charges for use are $12
for the first 10 minutes, $16/hr for longer periods.
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