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Automated analysis of septal fluorescence peak intensity

Grolman, Jeroen (2020) Automated analysis of septal fluorescence peak intensity. Master's Research Project 1, Biology.

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Abstract

Analysis of peptidoglycan and bacterial cell wall by fluorescence microscopy can teach us a lot about the cell growth dynamics and cell morphogenesis. Analysis of these images manually is time consuming. In this work, we present the comparison between manual data collection of fluorescence intensity for the dyes FM 4-64, Nile Red (which both stain membranes, accumulating more in fluid membrane regions) and HADA (which is incorporated into newly synthesized peptidoglycan, marking peptidoglycan synthesis activity) in Bacillus subtilis and a more automated method of data collection based on the ChainTracer plugin for FIJI. We also present an additional tool created on this software which allows the user to measure fluorescence profiles along the short axis (width) of the cell. Our data shows that for individual measurements the automatically collected data matches the manually collected data. Our data shows evidence of colocalization between HADA and FM 464 dyes, and between HADA and Nile Red, which suggests that new peptidoglycan synthesis is happening in the same place as septa formation.

Item Type: Thesis (Master's Research Project 1)
Supervisor name: Scheffers, D.J. and Lopes Leao, P.E.
Degree programme: Biology
Thesis type: Master's Research Project 1
Language: English
Date Deposited: 01 Sep 2020 13:17
Last Modified: 01 Sep 2020 13:17
URI: https://fse.studenttheses.ub.rug.nl/id/eprint/23348

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