My study, FACS and imaging of surface marker of Breast cancer cell lines, i used Ab conjugate with FITC. I used 4% PFA for 10 min for fixing . I have fitc bleaching most of the time. can I use 2% PFA/antibleaching solution to avoid FITC bleaching?. request suggestion. thanx in advance. Mahesh
Joseph Christakiran Moses
Ice cold methanol fixation could be tried! Add Ice cold methanol, incubate at -20 deg Celsius for 15 minutes.
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Benjamin Samuel Goldschmidt
I would suggest using quantum dots instead of FITC. Quantum dots are highly resistant to photobleaching compared to FITC.
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Sherif Suleiman
Its also best to avoid direct light. And maybe alexa conjugated abs give a brighter signal than fitc
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