Fluorometer Operation - Promega E8051 Technical Manual

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 Excitation Peak
Kits* Wavelength
UV 365nm
Blue 490nm
Green 525nm
Red 625nm
*Optical kits are named based on the color of the excitation wavelength.
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.
Revised 5/10
Emission
Wavelength Typical Fluorophores
410–460nm Hoechst dye, 4-MU
510–570nm EGFP, or hMGFP, PicoGreen
®
RiboGreen
iT™ Protein, Quant-iT™
dsDNA, Rhodamine-110
580–640nm Rhodamine, Cy
660–720nm Cy ® 5, Quant-iT™ RNA
® ,
, Fluorescein, Quant-
® 3, resorufin
Part# TM311
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