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Hi I have 2 questions1. For how many hours do I incubate...
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Parul Rai Apr 2, 2015
Hi I have 2 questions
1. For how many hours do I incubate hematopoietic stem cells after staining them?
2. I sort my stem cells directly onto a slide and wait for about 45 min to let them adhere to the poly-lysine slide prior to fixing them. Unfortunately I have lost the cells wh Read more
1. For how many hours do I incubate hematopoietic stem cells after staining them?
2. I sort my stem cells directly onto a slide and wait for about 45 min to let them adhere to the poly-lysine slide prior to fixing them. Unfortunately I have lost the cells wh Read more
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Caiyun Grace Li Author Answered Apr 2, 2015
Department of Radiation Oncology, Stanford University, USA
Hi Paul,
Here is the answer -
Q1: We do not have experience with stem cells. I would suggest you to monitor the color development anytime from 6h to 48h (or even longer) incubation, until you see blue colored cells.
Q2: Residual fixative (PFA) might affect staining and reproducibility. I would suggest to wash at least twice before proceed. However, I would suggest to coat dishes/slides with different reagents (fibronectin, collagen, or gelatin) to reduce detachment of the cells.
Hope this helps.
Here is the answer -
Q1: We do not have experience with stem cells. I would suggest you to monitor the color development anytime from 6h to 48h (or even longer) incubation, until you see blue colored cells.
Q2: Residual fixative (PFA) might affect staining and reproducibility. I would suggest to wash at least twice before proceed. However, I would suggest to coat dishes/slides with different reagents (fibronectin, collagen, or gelatin) to reduce detachment of the cells.
Hope this helps.
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