Abstract
High-pressure freezing limits the size of biological samples, because only small samples can be frozen without ice damage. Additionally, these samples must fit into the dimensions of the sample holder provided by the high-pressure freezer. We explored the potential of a 10 μm thin polyester filter membrane (PE-filter) as a versatile sample substrate for high-pressure freezing. Planktonic bacteria, bacterial spores and suspended eukaryotic cells could be concentrated on the PE-filter, whereas biofilm, bacterial microcolonies and HeLa cells were able to grow directly on the PE-filter. These microorganism-loaded PE-filters were used for high-pressure freezing, freeze-substitution and plastic embedding in Epon or Lowicryl. Embedded filters were cross-sectioned so that the interface between microorganism and substrate as well as the overlying medium was revealed. Although the structural preservation was good for thin samples and samples with lower water content, such as biofilms, adherent HeLa-cell cultures were likewise sufficiently preserved for transmission electron microscopy imaging. The fact that microorganism-loaded PE-filters could be also examined with confocal laser scanning fluorescence microscopy under fully hydrated con...Continue Reading
References
Oct 1, 1978·Thorax·D Heath, P Smith
Jan 1, 1990·Histochemistry·J RothK T Tokuyasu
Jun 1, 1969·The Biochemical Journal·J M Sterlini, J Mandelstam
Sep 1, 1995·Journal of Microscopy·D StuderM D Buschmann
Jul 1, 1994·Journal of Microscopy·H HohenbergM Müller
May 29, 1998·Biochimica Et Biophysica Acta·Y HirukawaS Tomino
Apr 1, 1963·The Journal of Cell Biology·E S REYNOLDS
Oct 1, 2003·Journal of Microscopy·M K Morphew, J R McIntosh
Oct 1, 2003·Journal of Microscopy·P MonaghanF Tomley
Mar 22, 2006·The Journal of Histochemistry and Cytochemistry : Official Journal of the Histochemistry Society·Paul WebsterJoseph W St Geme
Jan 27, 2009·Journal of Bacteriology·Hildur PalsdottirManfred Auer
Sep 28, 2010·Methods in Cell Biology·Michael Laue
Sep 28, 2010·Methods in Cell Biology·Kent McDonaldMary Morphew
Apr 15, 2014·Environmental Microbiology·Diego O Serra, Regine Hengge
Jan 1, 2014·BioTechniques·Ellen LambrechtKurt Houf
Dec 15, 2015·International Journal of Cell Biology·Xiaotang HuLynn Moscinski
Nov 25, 2018·Sensors·Kamila HrubanovaOta Samek