Multi-mode Plate Reader

iD3 multi-mode plate readerThe Biomolecular Analysis Facility has a Molecular Devices iD3 multi-mode (Fluorescence, luminescence, absorbance) plate reader. The instrument is located in  Pinn Hall room 1074, in the Biomolecular Analysis Facility.  Users need to contact Dr. JJ Park of the Biomolecular Analysis Facility at 434.924.0070 or by email at jp2ht@virginia.edu to establish an account on the computer which controls the instrument. Dr. Park will offer basic training for the operation of the instrument. Instructions to get started are available on the computer desktop. Most users need only a short time to take readings, so it is unusual to experience a wait to use the instrument, hence reservations are not required for standard use.

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 iD3 instrument are

  • dual monochromators (bandwidth 9 nm) allowing any fluorophore to be read within the range of 250 to 850 nm
  • luminescence range 300-850 nm
  • spectral scanning
  • top or bottom reading of plates for adherent cells
  • well scanning
  • kinetic measurements
  • 384, 96, 48, 24, 12, 6 well plates
  • heating of plates, 5° above ambient to 65°C
  • shaking of plate
  • SoftMax Pro 7.03 software for data analysis.
  • detection limit: for fluorescence; 1 pM fluorescein (top read)
  • luminescence dynamic range >7 decades, optimized sensitivity of 2 pM for ATP-Glow

This new instrument reads absorbance

It can perform some time resolved fluorescence measurements but not polarized fluorescence.

The new instrument  has injectors required for flash luminescence assays e.g. Promega Dual Glo Luciferase assays.

The instrument is user operated. The current charges for use are $15 for the first 10 minutes, $15/hr after the first 10 minutes.

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.