Pre-gelation staining expansion microscopy for visualisation of the Plasmodium liver stage.

Autor: Atchou K; Institute of Cell Biology, University of Bern, 3012 Bern, Switzerland.; Graduate School for Cellular and Biomedical Sciences, University of Bern, 3012 Bern, Switzerland., Berger BM; Institute of Cell Biology, University of Bern, 3012 Bern, Switzerland.; Graduate School for Cellular and Biomedical Sciences, University of Bern, 3012 Bern, Switzerland., Heussler V; Institute of Cell Biology, University of Bern, 3012 Bern, Switzerland., Ochsenreiter T; Institute of Cell Biology, University of Bern, 3012 Bern, Switzerland.
Jazyk: angličtina
Zdroj: Journal of cell science [J Cell Sci] 2023 Nov 15; Vol. 136 (22). Date of Electronic Publication: 2023 Nov 30.
DOI: 10.1242/jcs.261377
Abstrakt: Fluorescence and light microscopy are important tools in the history of natural science. However, the resolution of microscopes is limited by the diffraction of light. One possible method to circumvent this physical restriction is the recently developed expansion microscopy (ExM). However, the original ultrastructure ExM (U-ExM) protocol is very time-consuming, and some epitopes are lost during the process. In this study, we developed a shortened pre-gelation staining ExM (PS-ExM) protocol and tested it to investigate the Plasmodium liver stage. The protocol presented in this study allows expanding of pre-stained samples, which results in shorter incubation times, better preservation of some epitopes and the advantage that non-expanded controls can be performed alongside using the same staining protocol. The protocol applicability was accessed throughout the Plasmodium liver stage, showing isotropic five-fold expansion. Furthermore, we used PS-ExM to visualise parasite mitochondria as well as the association of lysosomes to the parasitophorous vacuole membrane (PVM) as an example of visualising host-pathogen interaction. We are convinced that this new tool will be helpful for a deeper understanding of the biology of the Plasmodium liver stage.
Competing Interests: Competing interests The authors declare no competing or financial interests.
(© 2023. Published by The Company of Biologists Ltd.)
Databáze: MEDLINE