Are centrifugations required between every wash step/blocking/antibody incubation in order to not lose the cells? Is there a way to make them adhere through a fixative or some other process in order to make the staining process easier? Thank you!
You can coat your slides with poly-L-lysine, let the cells decant and adhere and then perform the staining. I do tgis routinely for B-lymphocytes and despite some cells are lost throughout the process most of them will remain there in the end.
If you would like further info you could PM or email me, and I will be very glad to provide you as much help as needed.
Best,
D
Hello
Yes , Coat coverslips with polyethylineimine or poly-L-lysine for 1 h at room temperature.
See link here
http://www.abcam.com/protocols/immunocytochemistry-immunofluorescence-protocol
https://www.rndsystems.com/resources/protocols/protocol-preparation-and-fluorescent-icc-staining-non-adherent-cells
There are also some techniques that does not require fixative long term as the cytospin, but is a centrifuge for slides, normally people who study blood and monocytes use it. If not, the Poly-L-lysine coating would work as well.
Some people also spin the cells, add a fixative medium to the pellet but without pipetting (to preserve the pellet structure) and then they put the cells into a paraffin block and do sectioning.
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