I have difficulties to count the adipose tissue SVF cells using a Neubauer cell because there are very few cells and a lot of debris. What is the more efficient way to count these cells?
Thank you.
Hi Jack, have you tried regular H&E staining of your samples on regular slides.
H&E does not stain the lipid-bearing adipocytes but will stain the stromal cells. Take a look at these google images:
https://www.google.com/search?tbm=isch&sa=1&q=adipocyte+staining+H%26E&oq=adipocyte+staining+H%26E&gs_l=img.3...302774.311591.0.312166.4.4.0.0.0.0.144.436.2j2.4.0....0...1.1.64.img..0.1.141...0i24k1.a31ZEj7ahCs#imgrc=_&spf=1499614662056
Luna system using trypan blue is used classically for this purpose
If you ensure that your digestion protocol is optimised you shouldn't get too much debris. Excessive debris is usually a sign that the digestion is too aggressive. Also make sure you perform washing steps before and after digestion.
We use trypan blue and a haematocytometer and it works well.
Thank you very much for your answers ! Indeed I think that my digestion is not optimal, I will try to ameliorate my protocol.
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