Gel microdroplet technique leaving microorganisms alive for sorting by flow cytometry

Citation
T. Katsuragi et al., Gel microdroplet technique leaving microorganisms alive for sorting by flow cytometry, J MICROB M, 42(1), 2000, pp. 81-86
Citations number
6
Language
INGLESE
art.tipo
Article
Categorie Soggetti
Biology,Microbiology
Journal title
JOURNAL OF MICROBIOLOGICAL METHODS
ISSN journal
0167-7012 → ACNP
Volume
42
Issue
1
Year of publication
2000
Pages
81 - 86
Database
ISI
SICI code
0167-7012(200009)42:1<81:GMTLMA>2.0.ZU;2-3
Abstract
Fluorescent labels used to disclose cellular function and the like are gene rally needed for cytometric analysis, but suitable ones are not always avai lable, Thiamin, an example of such a label, was used as a model in this stu dy. Thiamin in cells of bakers' yeast can be chemically converted to thioch rome, which fluoresces strongly, but the reaction kills cells not protected inside a gel microdroplet (GMD). Our new procedure for preparation of a sm all amount of GMDs uses a glass filter with micropores uniform in size to m ake an emulsion of molten agar in mineral oil that is then chilled for gela tion of the agarose. Cells were suspended and mixed with the war-m agarose, which was used to make GMDs. Cells were grown into microcolonies in the GM Ds, which were then treated to convert thiamin to thiochrome. The thiochrom e in microcolonies in single GMDs could be detected by flow cytometry. Plat ing showed that enough cells survived for this method to be used for screen ing. This technique can be used for all applications of GMDs. (C) 2000 Else vier Science B.V. All rights reserved.