title: Targeted single-cell microchemical analysis: MS-based peptidomics of individual paraformaldehyde-fixed and immunolabeled neurons.
abstract: 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.
authors.affiliation: Department of Chemistry and the Beckman Institute for Advanced Science and Technology, University of Illinois Urbana-Champaign, Urbana, IL 61801, USA. mail@susanne-neupert.de
authors.forename: Susanne
authors.initials: S
authors.lastname: Neupert
authors.forename: Stanislav S
authors.initials: SS
authors.lastname: Rubakhin
authors.forename: Jonathan V
authors.initials: JV
authors.lastname: Sweedler
mesh_terms: D000818:Animals; D000906:Antibodies / Q000276:immunology; D002478:Cells, Cultured; D005557:Formaldehyde / Q000737:chemistry*; D007150:Immunohistochemistry; D009474:Neurons / Q000737:chemistry / Q000166:cytology / Q000378:metabolism*; D009479:Neuropeptides / Q000737:chemistry / Q000378:metabolism*; D010526:Periplaneta; D011108:Polymers / Q000737:chemistry*; D040901:Proteomics; D059010:Single-Cell Analysis; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
chemical_list: D000906:Antibodies; D009479:Neuropeptides; D011108:Polymers; D005557:Formaldehyde; C003043:paraform