Tissue sample preparation and MALDI MS imaging thereof
AssigneesBrandeis University, The Brigham and Women's Hospital, Inc.
InventorsJeffrey N. Agar, Nathalie Y. R. Agar
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Abstract
Aspects of the present invention relate to a method for the preparation of samples for MALDI MS imaging. Certain embodiments relate to a method of matrix deposition for samples, wherein tissue sections are prepared via a synergistic combination of fixation with matrix. In certain embodiments, tissue is fixed with cold solvent, according to well-established histology protocols, and in the presence of matrix, allowing for high resolution spatial mapping of protein, lipid, sugar, and/or nucleic acid distribution. In certain embodiments, the present invention relates to fixation with matrix of whole organisms. In certain embodiments, animals are perfused with fixation and matrix mixtures, which allows for direct mass spectrometry analysis.
Claims11
A method of generating an image of a sample, wherein the image details the spatial distribution and abundance of compounds in the sample, comprising:
- (a) combining:
- a fixative selected from the group consisting of:
- (i) acetone;
- (ii) methanol;
- (iii) about 95% ethanol and about 5% acetic acid;
- and (iv) 1 part methanol:1 part ethanol:0.5 parts acetonitrile:0.5 parts aqueous solution of about 0.1% TFA;
- with
- a mass spectrometry matrix material selected from the group consisting of:
- sinapinic acid and α-cyano-4-hydroxycinnamic acid,
- to give a fixative solution;
- (b) treating the sample selected from the group consisting of:
- a tissue sample, a cell culture, a single cell, a cellular extract, and a plurality of cells immobilized on a substrate surface, with said fixative solution, at a treating temperature from −78° C. to 37° C. and for a period of time sufficient to inhibit compounds in the sample from moving or diffusing from their original location therein, thereby forming a fixed sample;
- and
- (c) imaging the fixed sample using MALDI mass spectrometry, thereby generating the image of the sample.
Dependencies are inferred from the claim text.