I would like to use snap frozen tissue samples for proteome analysis in Mass Spectrometry Imaging but they have been emedded in Tissue-Tek during procurement. Due to certain circumstances and a level of rareness it is not possible to obtain these samples again anytime soon. Is there a possible method I can use to remove the Tissue-Tek and thus reduce the background signals that would otherwise interfere and dominate after analysis other then shaving it off with a scalpel?
Hi Anthony,
For MSI of proteins, you remove some Tissue-Tek with the EtOH wash. Of course, it is not complete removal, but could help a bit. Anyway, to get the best possible result, the precise scalpel skills will be required.
Btw, one of Caprioli's papers compares different washing methods... maybe you can find something there.
For MSI, you need to do serial washing anyway.
The vast majority of the published protocols uses a 3 step approach with serial washing in 70% EtOH, 95% EtOH and a final wash in a strong organic solvent (mostly isopropanol or choloroform).
A good part of your OCT is gone with the first 2 EtOH washing steps.
Peptide digest or intact protein imaging? The molecular ions will form with higher efficiency with less t-tek to ionize in the selected ionization matrix. If you seek peptide digests, Washing out t-tek with alcohols (what was mentioned above is standard) first will greatly help, but the MSI instrument ion source will still get dirty faster than with non-t-tek samples. That's an issue for the technician in charge of the Frequent cleaning needed for MSI ion sources.
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