Kevin Schey
Last active: 3/24/2020

A method to prevent protein delocalization in imaging mass spectrometry of non-adherent tissues: application to small vertebrate lens imaging.

Anderson DM, Floyd KA, Barnes S, Clark JM, Clark JI, Mchaourab H, Schey KL
Anal Bioanal Chem. 2015 407 (8): 2311-20

PMID: 25665708 · PMCID: PMC4379205 · DOI:10.1007/s00216-015-8489-5

MALDI imaging requires careful sample preparation to obtain reliable, high-quality images of small molecules, peptides, lipids, and proteins across tissue sections. Poor crystal formation, delocalization of analytes, and inadequate tissue adherence can affect the quality, reliability, and spatial resolution of MALDI images. We report a comparison of tissue mounting and washing methods that resulted in an optimized method using conductive carbon substrates that avoids thaw mounting or washing steps, minimizes protein delocalization, and prevents tissue detachment from the target surface. Application of this method to image ocular lens proteins of small vertebrate eyes demonstrates the improved methodology for imaging abundant crystallin protein products. This method was demonstrated for tissue sections from rat, mouse, and zebrafish lenses resulting in good-quality MALDI images with little to no delocalization. The images indicate, for the first time in mouse and zebrafish, discrete localization of crystallin protein degradation products resulting in concentric rings of distinct protein contents that may be responsible for the refractive index gradient of vertebrate lenses.

MeSH Terms (15)

Analytic Sample Preparation Methods Animals Crystallins Female Lens, Crystalline Male Mice Mice, Inbred C57BL Mice, Inbred ICR Molecular Imaging Protein Transport Rats Rats, Wistar Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization Zebrafish

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