Targeted Single-Cell Microchemical Analysis: MS-Based Peptidomics of Individual Paraformaldehyde-Fixed and Immunolabeled Neurons

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• 作者：Susanne Neupert ; Stanislav?S. Rubakhin ; Jonathan?V. Sweedler
• 刊名：Chemistry & Biology
• 出版年：2012
• 出版时间：24 August, 2012
• 年：2012
• 卷：19
• 期：8
• 页码：1010-1019
• 全文大小：675 K

文摘

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Summary

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.