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Combining Micropunch Histology and Multidimensional Lipidomic Measurements for In-Depth Tissue Mapping

[Image: see text] While decades of technical and analytical advancements have been utilized to discover novel lipid species, increase speciation, and evaluate localized lipid dysregulation at subtissue, cellular, and subcellular levels, many challenges still exist. One limitation is that the acquisi...

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Autores principales: Odenkirk, Melanie T., Horman, Brian M., Dodds, James N., Patisaul, Heather B., Baker, Erin S.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Chemical Society 2021
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9139744/
https://www.ncbi.nlm.nih.gov/pubmed/35647605
http://dx.doi.org/10.1021/acsmeasuresciau.1c00035
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author Odenkirk, Melanie T.
Horman, Brian M.
Dodds, James N.
Patisaul, Heather B.
Baker, Erin S.
author_facet Odenkirk, Melanie T.
Horman, Brian M.
Dodds, James N.
Patisaul, Heather B.
Baker, Erin S.
author_sort Odenkirk, Melanie T.
collection PubMed
description [Image: see text] While decades of technical and analytical advancements have been utilized to discover novel lipid species, increase speciation, and evaluate localized lipid dysregulation at subtissue, cellular, and subcellular levels, many challenges still exist. One limitation is that the acquisition of both in-depth spatial information and comprehensive lipid speciation is extremely difficult, especially when biological material is limited or lipids are at low abundance. In neuroscience, for example, it is often desired to focus on only one brain region or subregion, which can be well under a square millimeter for rodents. Herein, we evaluate a micropunch histology method where cortical brain tissue at 2.0, 1.25, 1.0, 0.75, 0.5, and 0.25 mm diameter sizes and 1 mm depth was collected and analyzed with multidimensional liquid chromatography, ion mobility spectrometry, collision induced dissociation, and mass spectrometry (LC-IMS-CID-MS) measurements. Lipid extraction was optimized for the small sample sizes, and assessment of lipidome coverage for the 2.0 to 0.25 mm diameter sizes showed a decline from 304 to 198 lipid identifications as validated by all 4 analysis dimensions (∼35% loss in coverage for ∼88% less tissue). While losses were observed, the ∼200 lipids and estimated 4630 neurons contained within the 0.25 punch still provided in-depth characterization of the small tissue region. Furthermore, while localization routinely achieved by mass spectrometry imaging (MSI) and single cell analyses is greater, this diameter is sufficiently small to isolate subcortical, hypothalamic, and other brain regions in adult rats, thereby increasing the coverage of lipids within relevant spatial windows without sacrificing speciation. Therefore, micropunch histology enables in-depth, region-specific lipid evaluations, an approach that will prove beneficial to a variety of lipidomic applications.
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spelling pubmed-91397442022-05-27 Combining Micropunch Histology and Multidimensional Lipidomic Measurements for In-Depth Tissue Mapping Odenkirk, Melanie T. Horman, Brian M. Dodds, James N. Patisaul, Heather B. Baker, Erin S. ACS Meas Sci Au [Image: see text] While decades of technical and analytical advancements have been utilized to discover novel lipid species, increase speciation, and evaluate localized lipid dysregulation at subtissue, cellular, and subcellular levels, many challenges still exist. One limitation is that the acquisition of both in-depth spatial information and comprehensive lipid speciation is extremely difficult, especially when biological material is limited or lipids are at low abundance. In neuroscience, for example, it is often desired to focus on only one brain region or subregion, which can be well under a square millimeter for rodents. Herein, we evaluate a micropunch histology method where cortical brain tissue at 2.0, 1.25, 1.0, 0.75, 0.5, and 0.25 mm diameter sizes and 1 mm depth was collected and analyzed with multidimensional liquid chromatography, ion mobility spectrometry, collision induced dissociation, and mass spectrometry (LC-IMS-CID-MS) measurements. Lipid extraction was optimized for the small sample sizes, and assessment of lipidome coverage for the 2.0 to 0.25 mm diameter sizes showed a decline from 304 to 198 lipid identifications as validated by all 4 analysis dimensions (∼35% loss in coverage for ∼88% less tissue). While losses were observed, the ∼200 lipids and estimated 4630 neurons contained within the 0.25 punch still provided in-depth characterization of the small tissue region. Furthermore, while localization routinely achieved by mass spectrometry imaging (MSI) and single cell analyses is greater, this diameter is sufficiently small to isolate subcortical, hypothalamic, and other brain regions in adult rats, thereby increasing the coverage of lipids within relevant spatial windows without sacrificing speciation. Therefore, micropunch histology enables in-depth, region-specific lipid evaluations, an approach that will prove beneficial to a variety of lipidomic applications. American Chemical Society 2021-10-28 /pmc/articles/PMC9139744/ /pubmed/35647605 http://dx.doi.org/10.1021/acsmeasuresciau.1c00035 Text en © 2021 The Authors. Published by American Chemical Society https://creativecommons.org/licenses/by-nc-nd/4.0/Permits non-commercial access and re-use, provided that author attribution and integrity are maintained; but does not permit creation of adaptations or other derivative works (https://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Odenkirk, Melanie T.
Horman, Brian M.
Dodds, James N.
Patisaul, Heather B.
Baker, Erin S.
Combining Micropunch Histology and Multidimensional Lipidomic Measurements for In-Depth Tissue Mapping
title Combining Micropunch Histology and Multidimensional Lipidomic Measurements for In-Depth Tissue Mapping
title_full Combining Micropunch Histology and Multidimensional Lipidomic Measurements for In-Depth Tissue Mapping
title_fullStr Combining Micropunch Histology and Multidimensional Lipidomic Measurements for In-Depth Tissue Mapping
title_full_unstemmed Combining Micropunch Histology and Multidimensional Lipidomic Measurements for In-Depth Tissue Mapping
title_short Combining Micropunch Histology and Multidimensional Lipidomic Measurements for In-Depth Tissue Mapping
title_sort combining micropunch histology and multidimensional lipidomic measurements for in-depth tissue mapping
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9139744/
https://www.ncbi.nlm.nih.gov/pubmed/35647605
http://dx.doi.org/10.1021/acsmeasuresciau.1c00035
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