Context:
- Isolated red blood cells (protocol in .docx is attached)
- Did confocal microscopy (stained RBCs with CellMask Red (photoiodide))
- Observed red blood cells (circled in blue) and also some cells with spiky surfaces (circled in green) (image as attached)
Questions:
- What are those spiky cells?
- Is it normal for my red blood cell extracts to have them?
- If not, did something go wrong in my sample preparation, and how do I get rid of them?
Thank you!!
Malcolm Nobre
Hello Si Ying Lim
After reading your protocol, I suppose you did not add cryopreservant to the RBCs when stored at -80 degree C.
Please refer to the paper below.
Red blood cell phenotype fidelity following glycerol cryopreservation optimized for research purposes.
Article Red blood cell phenotype fidelity following glycerol cryopre...
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Ellen A G Chernoff
The simplest explation is that your sample is drying out. As the buffer becomes more hypertonic RBCs undergo crenation. https://www.thoughtco.com/crenation-definition-and-example-609188
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