« Transformation Differences | Main | Comp Matrix file tips »

September 10, 2011

Comments

Shan Yu

Thanks so much for the tips on compensation. I have a question here. How do I know if a kind of compensation particle has the same auto-florescence as my samples before purchasing it?

vincent

Dear Nick,

you wrote:"Finally, compensation controls must be treated in the same manner as experimental samples. This is because exposure to light and treatments like fixation/permeabilization may alter the fluorochrome, particularly the tandem conjugation ratio, i.e. lose some Cy7 on each PE molecule. "

If we are fixing (formaldehyde) samples post-staining, and acquiring the following day, why not prepare "fresh" compensation beads (and without fixative solution treatment)? I thought that what was essential was to get a positive fluo peak that follows rule#1. if the fixative treatment is altering (lowering) the level of fluo, why couldn't we use comp beads freshly prepared giving higher fluo?

Many thanks,
Vincent

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