Fluorescent Dye Compatability Groups

In flow cytometry, there are a large number of dyes with excitation and emission profiles that are so similar that two dyes from the same group cannot be used together in the same experiment. The table below indicates such groups

Group Laser Dyes
Violet A 407 nm Cascade Blue, Alexa405, Pacific Blue
Violet B 407 nm

Alexa430, AmCyan, Qdot525

Blue A 488 nm

FITC, Fluorescein-NHS, Alexa488, GFP

Blue/Green B 488 / 532 nm PE
Blue/Green B 488 / 532 nm PE-TexasRed, PE-Alexa595, PE-Alexa610
Blue/Green C 488 / 532 nm

PE-Cy5, PE-Alexa647

Blue/Green D 488 / 532 nm PE-Cy5.5, PE-Alexa680, PE-Alexa700
Blue/Green E 488 / 532 nm PE-Cy7, PE-Alexa750
Red A 633 nm APC, Cy5, Alexa647
Red B 633 nm APC-Cy5.5, APC-Alexa680, APC-Alexa700, Cy5.5, Alexa680, Alexa700
Red C 633 nm APC-Cy7, APC-Alexa750

Quantum dots are an emerging class of reagents for flow cytometry and custom conjugation kits are now available from Quantum Dot Corporation. Unlike the dyes above, they have very broad excitation specta, and are usually used off of the Violet laser. Many labs are equipping their LSR-II cytometers with octagon detection blocks off the violet laser, and spacing out appropriate quantum dots accordingly. For facilities with standard trigon detectors off the violet laser, quantum dots could be used in place of the Violet B group, and a Violet C group could be added.