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Laser capture microdissection MALDI for direct analysis of archival tissue
Sucharita H Bhattacharya1, Anthony A Gal, Kermit K Murray
1Department of Chemistry, Louisiana State University, Baton Rouge, Louisiana 70803, USA.
Journal of Proteome Research
|March 20, 2003
Summary
Laser capture microdissection (LCM) enabled MALDI mass spectrometry analysis of lung cancer cells. Archived tissues yielded fewer peaks than fresh tissues, suggesting biomolecule degradation.
Area of Science:
- Biochemistry
- Oncology
- Analytical Chemistry
Background:
- Archived human lung tissues are valuable resources for cancer research.
- Matrix-assisted laser desorption/ionization (MALDI) mass spectrometry is a powerful tool for molecular analysis.
Purpose of the Study:
- To evaluate MALDI mass spectrometry for analyzing cancer cells isolated by laser capture microdissection (LCM) from archived lung tissues.
- To compare MALDI mass spectra from LCM-isolated cells with those from intact tissue sections.
Main Methods:
- Human lung adenocarcinoma and squamous cell carcinoma tissues were sectioned, stained, and dehydrated.
- Cancer cells were isolated using LCM, mixed with matrix, and analyzed by MALDI mass spectrometry.
- Intact tissue sections were also analyzed by MALDI mass spectrometry for comparison.
Main Results:
- MALDI mass spectra revealed 15-20 peaks between 1,000 and 4,000 Da, with approximately half unique to either cancer type.
- Direct MALDI analysis of tissue sections frozen in optimal cutting temperature (OCT) solution yielded optimal results.
- Fewer peaks were observed in archived tissues (both LCM and non-LCM) compared to fresh tissues, indicating potential biomolecule degradation.
Conclusions:
- MALDI mass spectrometry can analyze cancer cells isolated by LCM from archived tissues.
- Direct MALDI analysis of OCT-embedded tissue sections is a viable alternative to LCM.
- Biomolecule integrity in archived tissues may be compromised, impacting MALDI mass spectrometry results.