Intracellular Amyloid‑β Detection from Human Brain Sections Using a Microfluidic Immunoassay in Tandem with MALDI-MS
Alzheimer’s disease (AD) currently affects more than 30 million people worldwide. The lack of understanding of AD’s physiopathology limits the development of therapeutic and diagnostic tools. Soluble amyloid-β peptide (Aβ) oligomers that appear as intermediates along the Aβ aggregation into plaques...
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| Veröffentlicht in: | Analytical chemistry (Washington) 2023-04, Vol.95 (13), p.5522-5531 |
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| creator | Villarreal, Jorvani Cruz Kow, Keegan Pham, Brian Egatz-Gomez, Ana Sandrin, Todd R. Coleman, Paul D. Ros, Alexandra |
| description | Alzheimer’s disease (AD) currently affects more than 30 million people worldwide. The lack of understanding of AD’s physiopathology limits the development of therapeutic and diagnostic tools. Soluble amyloid-β peptide (Aβ) oligomers that appear as intermediates along the Aβ aggregation into plaques are considered among the main AD neurotoxic species. Although a wealth of data are available about Aβ from in vitro and animal models, there is little known about intracellular Aβ in human brain cells, mainly due to the lack of technology to assess the intracellular protein content. The elucidation of the Aβ species in specific brain cell subpopulations can provide insight into the role of Aβ in AD and the neurotoxic mechanism involved. Here, we report a microfluidic immunoassay for in situ mass spectrometry analysis of intracellular Aβ species from archived human brain tissue. This approach comprises the selective laser dissection of individual pyramidal cell bodies from tissues, their transfer to the microfluidic platform for sample processing on-chip, and mass spectrometric characterization. As a proof-of-principle, we demonstrate the detection of intracellular Aβ species from as few as 20 human brain cells. |
| doi_str_mv | 10.1021/acs.analchem.2c03825 |
| format | Article |
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The lack of understanding of AD’s physiopathology limits the development of therapeutic and diagnostic tools. Soluble amyloid-β peptide (Aβ) oligomers that appear as intermediates along the Aβ aggregation into plaques are considered among the main AD neurotoxic species. Although a wealth of data are available about Aβ from in vitro and animal models, there is little known about intracellular Aβ in human brain cells, mainly due to the lack of technology to assess the intracellular protein content. The elucidation of the Aβ species in specific brain cell subpopulations can provide insight into the role of Aβ in AD and the neurotoxic mechanism involved. Here, we report a microfluidic immunoassay for in situ mass spectrometry analysis of intracellular Aβ species from archived human brain tissue. This approach comprises the selective laser dissection of individual pyramidal cell bodies from tissues, their transfer to the microfluidic platform for sample processing on-chip, and mass spectrometric characterization. As a proof-of-principle, we demonstrate the detection of intracellular Aβ species from as few as 20 human brain cells.</description><identifier>ISSN: 0003-2700</identifier><identifier>ISSN: 1520-6882</identifier><identifier>EISSN: 1520-6882</identifier><identifier>DOI: 10.1021/acs.analchem.2c03825</identifier><identifier>PMID: 36894164</identifier><language>eng</language><publisher>United States: American Chemical Society</publisher><subject>Alzheimer Disease - metabolism ; Alzheimer's disease ; Amyloid beta-Peptides - chemistry ; analytical chemistry ; Animal models ; Animals ; Brain ; Brain - metabolism ; Chemistry ; dissection ; Humans ; Immunoassay ; immunoassays ; Intermediates ; Intracellular ; Mass spectrometry ; Mass spectroscopy ; Microfluidics ; Neurodegenerative diseases ; Neurotoxicity ; pathophysiology ; peptides ; Plaque, Amyloid - metabolism ; protein content ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization - methods ; Subpopulations ; Technology assessment ; therapeutics ; β-Amyloid</subject><ispartof>Analytical chemistry (Washington), 2023-04, Vol.95 (13), p.5522-5531</ispartof><rights>2023 The Authors. 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Chem</addtitle><description>Alzheimer’s disease (AD) currently affects more than 30 million people worldwide. The lack of understanding of AD’s physiopathology limits the development of therapeutic and diagnostic tools. Soluble amyloid-β peptide (Aβ) oligomers that appear as intermediates along the Aβ aggregation into plaques are considered among the main AD neurotoxic species. Although a wealth of data are available about Aβ from in vitro and animal models, there is little known about intracellular Aβ in human brain cells, mainly due to the lack of technology to assess the intracellular protein content. The elucidation of the Aβ species in specific brain cell subpopulations can provide insight into the role of Aβ in AD and the neurotoxic mechanism involved. Here, we report a microfluidic immunoassay for in situ mass spectrometry analysis of intracellular Aβ species from archived human brain tissue. This approach comprises the selective laser dissection of individual pyramidal cell bodies from tissues, their transfer to the microfluidic platform for sample processing on-chip, and mass spectrometric characterization. As a proof-of-principle, we demonstrate the detection of intracellular Aβ species from as few as 20 human brain cells.</description><subject>Alzheimer Disease - metabolism</subject><subject>Alzheimer's disease</subject><subject>Amyloid beta-Peptides - chemistry</subject><subject>analytical chemistry</subject><subject>Animal models</subject><subject>Animals</subject><subject>Brain</subject><subject>Brain - metabolism</subject><subject>Chemistry</subject><subject>dissection</subject><subject>Humans</subject><subject>Immunoassay</subject><subject>immunoassays</subject><subject>Intermediates</subject><subject>Intracellular</subject><subject>Mass spectrometry</subject><subject>Mass spectroscopy</subject><subject>Microfluidics</subject><subject>Neurodegenerative diseases</subject><subject>Neurotoxicity</subject><subject>pathophysiology</subject><subject>peptides</subject><subject>Plaque, Amyloid - metabolism</subject><subject>protein content</subject><subject>Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization - methods</subject><subject>Subpopulations</subject><subject>Technology assessment</subject><subject>therapeutics</subject><subject>β-Amyloid</subject><issn>0003-2700</issn><issn>1520-6882</issn><issn>1520-6882</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2023</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkU9u1DAUhy1ERYeBGyBkiQ2bDM__k-XQAh1pRl20XUeO7VBXcVLsRNXsuAJX6UF6CE6CRzPtgkW78uJ9v9-T34fQBwILApR80SYtdK87c-3CghpgJRWv0IwICoUsS_oazQCAFVQBHKO3Kd0AEAJEvkHHTJYVJ5LP0N2qH6M2ruumTke8DNtu8Pbv7z8P9_jUjc6MfuhxG4eAz6age_w1at_ji_0g4avk-59Y4403cWi7yVtv8CqEqR90SnqLM3ype-sCvvPjNd4s16erYnPxDh21ukvu_eGdo6vv3y5Pzor1-Y_VyXJdaEYqUVhJCK2EbhpJrOWWaFsSMFxRaChIRZUjhggBnFbKMslBgiMNb1olFC8Zm6PP-97bOPyaXBrr4NPuu7p3w5RqWjJORSbly6gqJVRMVCqjn_5Db4YpZhk7qqooZSoXzxHfU_k0KUXX1rfRBx23NYF657DODutHh_XBYY59PJRPTXD2KfQoLQOwB3bxp8XPdv4D5QuqmQ</recordid><startdate>20230404</startdate><enddate>20230404</enddate><creator>Villarreal, Jorvani Cruz</creator><creator>Kow, Keegan</creator><creator>Pham, Brian</creator><creator>Egatz-Gomez, Ana</creator><creator>Sandrin, Todd R.</creator><creator>Coleman, Paul D.</creator><creator>Ros, Alexandra</creator><general>American Chemical Society</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7QF</scope><scope>7QO</scope><scope>7QQ</scope><scope>7SC</scope><scope>7SE</scope><scope>7SP</scope><scope>7SR</scope><scope>7TA</scope><scope>7TB</scope><scope>7TM</scope><scope>7U5</scope><scope>7U7</scope><scope>7U9</scope><scope>8BQ</scope><scope>8FD</scope><scope>C1K</scope><scope>F28</scope><scope>FR3</scope><scope>H8D</scope><scope>H8G</scope><scope>H94</scope><scope>JG9</scope><scope>JQ2</scope><scope>KR7</scope><scope>L7M</scope><scope>L~C</scope><scope>L~D</scope><scope>P64</scope><scope>7X8</scope><scope>7S9</scope><scope>L.6</scope><orcidid>https://orcid.org/0000-0002-6080-943X</orcidid><orcidid>https://orcid.org/0000-0001-7709-8331</orcidid></search><sort><creationdate>20230404</creationdate><title>Intracellular Amyloid‑β Detection from Human Brain Sections Using a Microfluidic Immunoassay in Tandem with MALDI-MS</title><author>Villarreal, Jorvani Cruz ; 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Here, we report a microfluidic immunoassay for in situ mass spectrometry analysis of intracellular Aβ species from archived human brain tissue. This approach comprises the selective laser dissection of individual pyramidal cell bodies from tissues, their transfer to the microfluidic platform for sample processing on-chip, and mass spectrometric characterization. As a proof-of-principle, we demonstrate the detection of intracellular Aβ species from as few as 20 human brain cells.</abstract><cop>United States</cop><pub>American Chemical Society</pub><pmid>36894164</pmid><doi>10.1021/acs.analchem.2c03825</doi><tpages>10</tpages><orcidid>https://orcid.org/0000-0002-6080-943X</orcidid><orcidid>https://orcid.org/0000-0001-7709-8331</orcidid><oa>free_for_read</oa></addata></record> |
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| subjects | Alzheimer Disease - metabolism Alzheimer's disease Amyloid beta-Peptides - chemistry analytical chemistry Animal models Animals Brain Brain - metabolism Chemistry dissection Humans Immunoassay immunoassays Intermediates Intracellular Mass spectrometry Mass spectroscopy Microfluidics Neurodegenerative diseases Neurotoxicity pathophysiology peptides Plaque, Amyloid - metabolism protein content Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization - methods Subpopulations Technology assessment therapeutics β-Amyloid |
| title | Intracellular Amyloid‑β Detection from Human Brain Sections Using a Microfluidic Immunoassay in Tandem with MALDI-MS |
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