Mapping of surrogate markers of cellular components and structures using laser desorption/ionization mass spectrometry.
Laser desorption/ionization mass spectrometry (LDI-MS) has been used to assess the potential of using surrogate markers, bound to cellular structures containing nucleic acids, to image or map the position of these structures within biological samples. In this study, organic dyes were used as markers because of their established use in the histochemical marking of nucleic acids, and also because they are amenable to LDI-MS. Eight cationic dyes were tested and all could be desorbed from nucleic acid samples without additional matrix after specifically binding to these molecules. Methylene Blue was the best of these based on its sensitivity to detection by LDI-MS and the fact that it can be washed from the tissue in areas where it was not specifically bound to provide low-intensity background signals. Experiments are reported which characterize the M(+) ion signal obtained from Methylene Blue with regard to sensitivity, reproducibility and possible use for quantitation. This dye was used to map (with a lateral resolution of 25 microm) several nucleic acid-containing samples spotted on prepared surfaces, and to image the location of nucleic acids in two model tissues, retinal vertical sections and thyroid whole mount sections.
Koomen
J M
JM
Laboratory for Biological Mass Spectrometry, Chemistry Department, Texas A&M University, College Station, Texas 77842, USA.
Stoeckli
M
M
Caprioli
R M
RM
D000818:Animals; D015415:Biomarkers; D002477:Cells / Q000737:chemistry*; D004396:Coloring Agents / Q000737:chemistry; D004247:DNA / Q000737:chemistry; D008751:Methylene Blue / Q000737:chemistry; D008856:Microscopy, Fluorescence; D009696:Nucleic Acids / Q000032:analysis; D011817:Rabbits; D051381:Rats; D012160:Retina / Q000737:chemistry / Q000166:cytology; D012441:Saccharomyces cerevisiae / Q000737:chemistry; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013961:Thyroid Gland / Q000737:chemistry / Q000166:cytology
D015415:Biomarkers; D004396:Coloring Agents; D009696:Nucleic Acids; D004247:DNA; D008751:Methylene Blue