Naslov
Light sheet fluorescence microscopy in the assessment of structural changes in the mouse brain after ischemic lesion

Autori
Hamer, Dominik ; Petrinec, Daniela ; Gajović, Srećko

Vrsta, podvrsta i kategorija rada
Sažeci sa skupova, sažetak, znanstveni

Izvornik
E-book of abstracts / - Zagreb, 2022, 99-99

Skup
8th Croatian Neuroscience Congress

Mjesto i datum
Zagreb, Hrvatska, 24.09.2021. - 25.09.2021

Vrsta sudjelovanja
Poster

Vrsta recenzije
Domaća recenzija

Ključne riječi
light sheet fluorescence microscopy (LSFM), mouse brain, tissue clearing, PEGASOS, iDISCO

Sažetak
Ischemic stroke is one of the major health problems in modern society. It occurs due to the blockage of blood vessels in the brain, which interrupts nutrients and oxygen supply to certain brain regions. Use of genetically modified animals is necessary, as they are irreplaceable and valuable models in preclinical research of ischemic stroke. Modern neuroimaging techniques provide insight in lesion localization and volume, but not in cell structure, dynamics and regeneration. Novel LSFM can be used on whole mouse brain in order to visualise cell morphology and molecular changes after stroke. Achieving high optical transparency is crucial step before using LSFM. The aim of this study is to establish tissue clearing method that provides good transparency for acquiring three-dimensional image of whole mouse brain using LSFM. Acquired 3D images could then be potentially correlated with other neuroimaging methods (MRI and in vivo bioluminescence) for better understanding of ischemic stroke by multimodal approach. Roughly, three types of tissue clearing techniques can be distinguished: organic-solvent-based, aqueous- based and hydrogel-based. Brains of adult wild type and Gap43-GFP mice were isolated and cleared using two organic-solvent-based methods: PEGASOS and iDISCO. Tissue transparency was estimated using grid drawn on a piece of paper. Brains were photographed after each tissue clearing step. Both blurriness and shrinkage of cleared brain tissue were less noticeable when using PEGASOS method. Autofluorescence was visualised on cleared brain slices using EVOS microscope. Brains cleared with PEGASOS method show satisfying transparency and could potentially be used for 3D image acquisition on LSFM.

Izvorni jezik
Engleski

Znanstvena područja
Temeljne medicinske znanosti

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