Bhattacharya Sucharita H, Gal Anthony A, Murray Kermit K
Department of Chemistry, Louisiana State University, Baton Rouge, Louisiana 70803, USA.
J Proteome Res. 2003 Jan-Feb;2(1):95-8. doi: 10.1021/pr025547m.
MALDI mass spectra were obtained from cancer cells isolated by laser capture microdissection (LCM) of archived tissue. Frozen human lung tissue from adenocarcenoma and squamous cell carcenoma cases were cut into 5 to 15 microm thick sections, stained with hematoxylin and dehydrated. Cancer cells were isolated by LCM, mixed with matrix solution, and deposited on a MALDI target for mass spectrometric analysis. For comparison with LCM isolated cells, tissue sections were placed directly on the MALDI target without microdissection. Tissue sections frozen in optimal cutting temperature (OCT) solution and cut into 8 microm thick sections gave the best performance with direct MALDI analysis. Between 15 and 20 peaks were observed in the mass region between 1,000 and 4,000 Da, and roughly half of these peaks were common to either squamous cells or adenocarcenoma. Additional peaks were observed in the non-LCM mass spectra and these may result from biomolecules in the healthy tissue. When compared to fresh tissue, both LCM and non-LCM archived tissue produced fewer peaks, possibly due to degradation of the biomolecules in the archived tissue.
基质辅助激光解吸电离质谱（MALDI）图谱是通过对存档组织进行激光捕获显微切割（LCM）分离得到的癌细胞获得的。将来自腺癌和鳞状细胞癌病例的冷冻人肺组织切成5至15微米厚的切片，用苏木精染色并脱水。通过LCM分离癌细胞，与基质溶液混合，并沉积在MALDI靶上进行质谱分析。为了与LCM分离的细胞进行比较，将组织切片直接放在MALDI靶上而不进行显微切割。冷冻在最佳切割温度（OCT）溶液中并切成8微米厚切片的组织切片在直接MALDI分析中表现最佳。在1000至4000 Da的质量区域中观察到15至20个峰，其中大约一半的峰在鳞状细胞或腺癌中是常见的。在非LCM质谱图中观察到其他峰，这些峰可能来自健康组织中的生物分子。与新鲜组织相比，LCM和非LCM存档组织产生的峰较少，这可能是由于存档组织中生物分子的降解。
