Imaging and mapping of tissue constituents at the single-cell level using maldi msi and quantitative laser scanning cytometry

2Citations
Citations of this article
10Readers
Mendeley users who have this article in their library.
Get full text

Abstract

For nearly a century, histopathology involved the laborious morphological analyses of tissues stained with broad-spectrum dyes (i.e., eosin to label proteins). With the advent of antibody-labeling, immunostaining (fluorescein and rhodamine for fluorescent labeling) and immunohistochemistry (DAB and hematoxylin), it became possible to identify specific immunological targets in cells and tissue preparations. Technical advances, including the development of monoclonal antibody technology, led to an ever-increasing palate of dyes, both fluorescent and chromatic. This provides an incredibly rich menu of molecular entities that can be visualized and quantified in cells-giving rise to the new discipline of Molecular Pathology. We describe the evolution of two analytical techniques, cytometry and mass spectrometry, which complement histopathological visual analysis by providing automated, cellular-resolution constituent maps. For the first time, laser scanning cytometry (LSC) and matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI-MSI) are combined for the analysis of tissue sections. The utility of the marriage of these techniques is demonstrated by analyzing mouse brains with neuron-specific, genetically encoded, fluorescent proteins. We present a workflow that: (1) can be used with or without expensive matrix deposition methods, (2) uses LSC images to reveal the diverse landscape of neural tissue as well as the matrix, and (3) uses a tissue fixation method compatible with a DNA stain. The proposed workflow can be adapted for a variety of sample preparation and matrix deposition methods.

Cite

CITATION STYLE

APA

Rawlins, C. M., Salisbury, J. P., Feldman, D. R., Isim, S., Agar, N. Y. R., Luther, E., & Agar, J. N. (2015). Imaging and mapping of tissue constituents at the single-cell level using maldi msi and quantitative laser scanning cytometry. In Methods in Molecular Biology (Vol. 1346, pp. 133–149). Humana Press Inc. https://doi.org/10.1007/978-1-4939-2987-0_10

Register to see more suggestions

Mendeley helps you to discover research relevant for your work.

Already have an account?

Save time finding and organizing research with Mendeley

Sign up for free