lately i have been trying to compare some erythrocytes in zebrafish larvaes, for this purpose we chose to do a wright giemsa stain. the problem came when we noticed that there where a lot of debris sticking to the slide, that gets stained.
Also , does anyone know a good fixation protocol for this cell type?, since we have already tried to fixate them with MetOH, EtOH and PFA 4%, and thus far we still get a lot of damaged cells.
I just have some suggestion because I have never worked with zebrafish.
- Fore your slides, before use, wash them with %70 ethanol and DW.
- Fore human samples we use EDTA, Did you use it?
Regards
thanks for your reply.
i can try the cleaning with EtoH, perhaps it could work.
for the EDTA part, im using HBSS (Hanks balanced salt solution) as an extraction medium, currently we are using a modified version of it with 1% EDTA V/V. it did help to improved the quantity of extracted erythrocytes, but we are still having problems ith the debris (wich we still suspect that it comes from the larvaes).
Best regards
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