I tried to HE staining white adipose tissue (Wistar rat). However, the membrane has broken, and the appearance is not right. My protocol is similar the found in Methods in Enzymology (MIE): Methods of Adipose Tissue. So could you help me with protocol suggestions?
to preserve tissue integrity, I used to stain adipose tissue on positively charged glasses!
Hi Joao, The adipose tissue is very delicate and sections are difficult to stretch without damaging them. I would suggest that you prepare a 10% alcohol (ethanol or IMS) in distilled water and pipette a large droplet of this solution on a positively charged slide (if you wish to collect serial sections - prepare a row of such slides). Place the adipose tissue section on the surface of the droplet. The surface tension of the droplet unfurls creases of the section and that allows for the next step - careful lowering of the slide into a water-bath (keep the temperature above 45 deg C, but you need to experiment to find the best temperature which gives you enough time for collecting the section without rushing (depends on your thermostat and the type of wax you are using). Watch the wax becoming more translucent and immediately collect it on the slide.
Until you are used to this method - it would be better to collect one section at a time as it is easy to miss the best moment for collect a section from the water-bath.
Good luck with your project.
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