期刊
JOURNAL OF FUNGI
卷 6, 期 4, 页码 -出版社
MDPI
DOI: 10.3390/jof6040257
关键词
fixation; scanning electron microscopy; matrix-assisted laser desorption; ionization mass spectrometry imaging; bacteria; fungi; rat lung tissue
资金
- Ministry of Education, Youth and Sports of the Czech Republic [LO1509]
- Czech Science Foundation [19-10907S]
A procedure for processing frozen rat lung tissue sections for scanning electron microscopy (SEM) from deeply frozen samples initially collected and stored for matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI-MSI) was developed. The procedure employed slow thawing of the frozen sections while floating on the surface and melting in a fixative solution. After the float-washing step, the sections were dehydrated in a graded ethanol series and dried in a critical point dryer. The SEM generated images with well-preserved structures, allowing for monitoring of bacterial cells and fungal hyphae in the infected tissue. Importantly, the consecutive nonfixed frozen sections were fully compatible with MALDI-MSI, providing molecular biomarker maps of Pseudomonas aeruginosa. The protocol enables bimodal image fusion in the in-house software CycloBranch, as demonstrated by SEM and MALDI-MSI.
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