Promega Corporation ·
2800 Woods Hollow Road
·
Madison, WI 53711-5399 USA
Toll Free in USA 800-356-9526
·
Phone 608-274-4330
·
Fax 608-277-2516
· www.promega.com
Printed in USA.
Part# TM311
Revised 5/10
Page 43
13. Fluorometer Operation
13.A. General Information
Microplate Recommendations
•
Black plates are highly recommended.
•
Clear plates can be used as a low-cost alternative if a high level of signal
intensity is expected.
•
White plates are
not
recommended. They will increase noise by 20–50
times.
Notes:
1. Samples with bubbles or high surface tension will affect fluorescence
readings.
2. Ensure the correct Optical Kit is used for the assay fluorophore.
3. If the instrument includes a Luminescence Module, it has a Microplate
Sample Tray Cover installed. The Microplate Sample Tray Cover is not
required for using fluorescence mode. To remove the Microplate Sample
Tray Cover, refer to Section 20.D for instructions.
4. All values recorded are in raw RFUs (relative fluorescence units). To
normalize, use the Curve-Fitting Data Analysis Software.
13.B. Optical Wavelengths and Commonly Used Fluorophores
Table 3. Commonly Used Fluorophores.
*Optical kits are named based on the color of the excitation wavelength.
Optical
Kits*
Excitation Peak
Wavelength
Emission
Wavelength
Typical Fluorophores
UV
365nm
410–460nm
Hoechst dye, 4-MU
Blue
490nm
510–570nm
EGFP, or hMGFP, PicoGreen
®
,
RiboGreen
®
, Fluorescein, Quant-
iT™ Protein, Quant-iT™
dsDNA, Rhodamine-110
Green
525nm
580–640nm
Rhodamine, Cy
®
3, resorufin
Red
625nm
660–720nm
Cy
®
5, Quant-iT™ RNA
