CyTRAK Orange™ is an excellent counterstain for your HCS experiments.
It's live-cell permeant which means it can be used in fixed or live end-point experiments.
Peak absorbance is at 510 nm meaning it can be excited with 488 nm, blue light.
It is spectrally separated from GFP and it is NOT excited by UV or 633/647 nm red light.
AND, it gives a nice nuclear:cyto segmentation from one channel for additional morphometrics.
Not to mention water-soluble and ready-to-use direct from the fridge!
Here's the evidence:
CYTRAK Orange™ as nucl:cyto counterstain was combined with AlexaFluor 350, AlexaFluor 633 labelled antibodies and GFP on the PE Operetta. This shows its compatibility with UV, blue and red excited fluors, while providing the nucl:cyto segmentation. Poster presentation: Sawada, J. et al., Univ. Shizuoka, Japan. Phenotype-based Screening of Chemical Library for Mitotic Inhibitors. High Content Analysis, Cambridge Healthtech, Inc.. San Francisco, January 2011.
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