Bringing SEM and MSI closer than ever before: Visualizing aspergillus and pseudomonas infection in the rat lungs

0536784 - MBÚ 2021 RIV CH eng J - Článek v odborném periodiku
Juříková, Tereza - Luptáková, Dominika - Kofroňová, Olga - Škríba, Anton - Novák, Jiří - Marešová, Helena - Palyzová, Andrea - Petřík, M. - Havlíček, Vladimír - Benada, Oldřich
Bringing SEM and MSI closer than ever before: Visualizing aspergillus and pseudomonas infection in the rat lungs.
Journal of Fungi. Roč. 6, č. 4 (2020), č. článku 257. E-ISSN 2309-608X
Grant CEP: GA MŠMT(CZ) LO1509; GA ČR(CZ) GA19-10907S
Institucionální podpora: RVO:61388971
Klíčová slova: fixation * scanning electron microscopy * matrix-assisted laser desorption/ionization mass spectrometry
Obor OECD: Microbiology
Impakt faktor: 5.816, rok: 2020
Způsob publikování: Open access
https://www.mdpi.com/2309-608X/6/4/257

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.
Trvalý link: http://hdl.handle.net/11104/0314544