Which is the best way to adhere cells on slide/coverslips?
I need to attach T cells on slide to make immunofluorecence assays and microscopy analysis
I don t have this material and i would prefere to let the cells settle down on slide but thanks for your interest and if you know other technics to make cells to adhere let me know. Meaning also the kind of coating that would be the best.
Cytospin is the best way to go. If you don't have cytospin, then try gentle smear and air dry. Then store the slides in storage buffer.
Specimen storage Buffer:
Dissolve 42.8g of sucrose (250mM) and 0.7g of MgCl2.6H2O (7mM)in 250 ml 1X PBS
Add 250 ml of Glycerol (50%) and mix well by stirring
Store at RT
Specimens/slides should be dipped and stored in -20oC
As an alternative to a cytospin-based method, I used to attach T cells to coverslips as below:
1. Wash cells 1x with D-PBS.
2. Resuspend cells in D-PBS.
3. Put the cell suspension onto a poly-L-lysine-coated coverslip.
4. Let the cells settle onto the coverslip for 10-20 mins @ 37C.
5. Remove excess cell suspension (check if enough cells are attached to the coverslip under microscope).
-> continue with fixation etc..
You can easily buy a ready to use poly-L-lysine solution from a company (e.g. Sigma) and coat coverslips with the chemical yourself.
Hope this helps your experiment.
Thanks a lot for all these answers i'll try evry possibility that you sent me.
best regards.
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