Details zur Publikation

Kategorie Textpublikation
Referenztyp Zeitschriften
DOI 10.1016/j.mimet.2008.05.017
Titel (primär) Fixation procedures for flow cytometric analysis of environmental bacteria
Autor Günther, S.; Hübschmann, T.; Rudolf, M.; Eschenhagen, M.; Röske, I.; Harms, H.; Müller, S.
Quelle Journal of Microbiological Methods
Erscheinungsjahr 2008
Department UMB
Band/Volume 75
Heft 1
Seite von 127
Seite bis 134
Sprache englisch
Keywords Fixation procedures; Flow cytometry; Bacterial stability; Single cell analysis; Equivalence testing; Bacterial DNA; Fluorescence techniques
Abstract Analysis of environmental bacteria on the single cell level often requires fixation to store the cells and to keep them in a state as near life-like as possible. Fixation procedures should furthermore counteract the increase of autofluorescence, cell clogging, and distortion of surface characteristics. Additionally, they should meet the specific fixation demands of both aerobically and anaerobically grown bacteria.A fixation method was developed based on metal solutions in combination with sodium azide. The fixation efficiencies of aluminium, barium, bismuth, cobalt, molybdenum, nickel, and tungsten salts were evaluated by flow cytometric measurement of the DNA contents as a bacterial population/community stability marker. Statistical equivalence testing was involved to permit highly reliable flow cytometric pattern evaluation. Investigations were carried out with pure cultures representing environmentally important metabolic and respiratory pathways as controls and with activated sludge as an example for highly diverse bacterial communities.A mixture of 5 mM barium chloride and nickel chloride, each and 10% sodium azide was found to be a suitable fixative for all tested bacteria. The described method provided good sample stability for at least 9 days.
dauerhafte UFZ-Verlinkung https://www.ufz.de/index.php?en=20939&ufzPublicationIdentifier=980
Günther, S., Hübschmann, T., Rudolf, M., Eschenhagen, M., Röske, I., Harms, H., Müller, S. (2008):
Fixation procedures for flow cytometric analysis of environmental bacteria
J. Microbiol. Methods 75 (1), 127 - 134 10.1016/j.mimet.2008.05.017