TY - JOUR
T1 - Targeted single-cell microchemical analysis
T2 - MS-based peptidomics of individual paraformaldehyde-fixed and immunolabeled neurons
AU - Neupert, Susanne
AU - Rubakhin, Stanislav S.
AU - Sweedler, Jonathan V.
N1 - Funding Information:
We acknowledge the financial assistance of the German Academic Exchange Service (DAAD; SN D/09/51168). The project described was supported by Award No. 5RO1NS031609 from the National Institute of Neurological Disorders and Stroke and Award Number P30 DA018310 from the National Institute on Drug Abuse. The content is solely the responsibility of the authors and does not necessarily represent the official views of the awarding agencies. We thank Professor M. Stengl (University of Kassel, Germany) and Xiying Wang (University of Illinois at Urbana-Champaign, USA) for help with cell culture and Professor R. Predel (Biocenter, University of Cologne, Germany) and Dr. M. Eckert (Friedrich-Schiller University, Jena, Germany) for providing the anti-Pea-PVK-II and anti-RFamide sera. S.N. conceived the project. S.N. performed experiments while getting appropriate advice from S.S.R. and J.V.S. S.N., S.S.R., and J.V.S. wrote the article.
PY - 2012/8/24
Y1 - 2012/8/24
N2 - Pinpointing a specific cell from within a relatively uniform cell population to determine its chemical content presents a challenging bioanalytical task. Immunocytochemistry is the classical method used to localize specific molecules and, hence, selected cells. Mass spectrometry also probes endogenous molecules such as neuropeptides within a cell. Here, these two approaches are hyphenated to allow microchemical analysis of immunocytochemical-selected peptidergic neurons. This two-step strategy utilizes antibody-based localization of cells containing selected biomarkers to isolate the cell(s) of interest, followed by peptidomic analysis via mass spectrometry. Applicable to a broad range of analyte and cell types, the strategy was used to successfully profile neuropeptides from individual immunostained insect neurons stored for up to 2 weeks as well as from tissues preserved for 42 weeks.
AB - Pinpointing a specific cell from within a relatively uniform cell population to determine its chemical content presents a challenging bioanalytical task. Immunocytochemistry is the classical method used to localize specific molecules and, hence, selected cells. Mass spectrometry also probes endogenous molecules such as neuropeptides within a cell. Here, these two approaches are hyphenated to allow microchemical analysis of immunocytochemical-selected peptidergic neurons. This two-step strategy utilizes antibody-based localization of cells containing selected biomarkers to isolate the cell(s) of interest, followed by peptidomic analysis via mass spectrometry. Applicable to a broad range of analyte and cell types, the strategy was used to successfully profile neuropeptides from individual immunostained insect neurons stored for up to 2 weeks as well as from tissues preserved for 42 weeks.
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U2 - 10.1016/j.chembiol.2012.05.023
DO - 10.1016/j.chembiol.2012.05.023
M3 - Article
C2 - 22921068
AN - SCOPUS:84865500786
SN - 2451-9448
VL - 19
SP - 1010
EP - 1019
JO - Cell Chemical Biology
JF - Cell Chemical Biology
IS - 8
ER -