We are trying to image mouse lymphatic vasculature by whole mount immunofluorescence, and these vessels are frequently associated with adipose tissue. The adipocytes obscure the view of the vessels when the samples are mounted. Physically dissecting away the fatty tissue is laborious, and can cause tearing or stretching of the tissue of interest. Are there any good ways of treating the sample, e.g. with an organic solvent, to get rid of adipocytes?
You could try using 2:1 Chloroform:Methanol. That should break open the adipocytes although I am not certain of the impact on the integrity of your samples for immunofluorescence.
Good Luck
If you use cloroform/methanol or other organic solvents, probably the lipids from the vasculature will be extracted together with those of the adipose. In my opinion, physical dissection under a magnifier is the best option.
The opinion in the literature is that alcohol fixation doesn't interfere with IF (at least with older fluorophores), so using chloroform/Methanol ensures fixation and clearing adipose tissue at the same time. Fixation is necessary for both integrity of tissue and retaining spacial distribution. Some years ago there was a method of preparation of tissue for breast cancer metasases assessment in axillary lymph nodes. As axilla contains a lot of adipose tissue - 'lymph node clearance' was obtained clearing whole surgical specimen with xylene. The specimens were fixed in 10% buffered formalin first. (could be done with xylene substitute). Good luck with your experiments.
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