Direct analysis of laser capture microdissected cells by MALDI mass spectrometry.
Laser capture microdissection (LCM) has become an important tool in biological research, permitting isolation of specific cell populations from frozen tissue samples containing a mixture of cell types. Cells obtained by LCM can be directly analyzed by matrix-assisted laser desorption/ionization mass spectrometry (MALDI MS). We report here methodology for the preparation and analysis of LCM captured cells with MALDI MS, giving high sensitivity and mass resolution. Comparison of the spectra obtained from cell populations of interest can identify unique disease or function-related protein markers. Using this approach, mass spectra obtained from human breast tissue containing invasive mammary carcinoma and normal breast epithelium using LCM were compared. Over 40 peaks were identified that significantly differed in intensity between invasive mammary carcinoma and normal breast epithelium. In addition, mass spectra are presented that show protein patterns from mouse liver and mouse colon crypts. The reported tissue preparation procedure and subsequent analysis by MALDI MS provide a new methodology for protein discovery involving LCM captured cells.
Xu
Baogang J
BJ
Department of Chemistry, Vanderbilt University, Nashville, Tennessee, USA.
Caprioli
Richard M
RM
Sanders
Melinda E
ME
Jensen
Roy A
RA
D000818:Animals; D001940:Breast / Q000737:chemistry / Q000166:cytology; D001943:Breast Neoplasms / Q000737:chemistry / Q000473:pathology; D002477:Cells / Q000737:chemistry*; D003106:Colon / Q000737:chemistry / Q000166:cytology / Q000473:pathology; D005260:Female; D006801:Humans; D007834:Lasers; D008099:Liver / Q000737:chemistry / Q000166:cytology / Q000473:pathology; D051379:Mice; D036103:Nanotechnology; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization