I am facing some issue regarding the sample preparation of FFPE tissue for the MALDI mass spectrometry Imaging.
As many protocols suggest I tried pulse boiling the slides in a citric acid solution in a microwave for 10 min. Unfortunately because of heat and air bubbles the tissue sections came off during this step.
I already tried some less harsh methods, by heating in the same solution on a heating block (no bubbles, lower temperature) but judging the spectra of these samples the antigen retrieval did not seem to work properly here.
I am happy to receive some tipps and hear from your experience.
Hi Charlotte,
If your fixation and embedding is alright, then the issue can potentially be addressed by the following:
. Ensure that you have good blades in the microtome thereby avoiding scratches, tears, etc. on the tissue
. You can coat the ITO slides with roughly 10-20 ul of poly-lysine in NP40 and dry it at 50oC before mounting the sections
. Heat the slides at 55-60oC for 1 hour immediately after mounting
Hope this helps.
Hi, I think fixing at 60C as suggested by Shibojyoti should solve your problem. Also, how thick sections do you use? Thicker sections tend to peel off easier.
Yes, it does work now - thank you for your answers! Lukas Krasny FYI: sections are 6 µm thick.
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