High-resolution imaging of pelagic bacteria by Atomic Force Microscopy and implications for carbon cycling

The ISME Journal
Francesca MalfattiFarooq Azam

Abstract

In microbial oceanography, cell size, volume and carbon (C) content of pelagic bacteria and archaea ('bacteria') are critical parameters in addressing the in situ physiology and functions of bacteria, and their role in the food web and C cycle. However, because of the diminutive size of most pelagic bacteria and errors caused by sample fixation and processing, an accurate measurement of the size and volume has been challenging. We used atomic force microscopy (AFM) to obtain high-resolution images of pelagic bacteria and Synechococcus. We measured the length, width and height of live and formalin-fixed pelagic bacteria, and computed individual cell volumes. AFM-based measurements were compared with those by epifluorescence microscopy (EFM) using 4',6-diamidino-2-phenylindole (DAPI). The ability to measure cell height by AFM provides methodological advantage and ecophysiological insight. For the samples examined, EFM (DAPI)-based average cell volume was in good agreement (1.1-fold) with live sample AFM. However, the agreement may be a fortuitous balance between cell shrinkage due to fixation/drying (threefold) and Z-overestimation (as EFM does not account for cell flattening caused by sample processing and assumes that height=wi...Continue Reading

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Citations

Feb 18, 2011·The ISME Journal·Daniel SherSallie W Chisholm
Jul 27, 2010·Applied and Environmental Microbiology·Melissa Garren, Farooq Azam
Sep 29, 2011·Applied and Environmental Microbiology·Emiley A EloeDouglas H Bartlett
Oct 17, 2012·PloS One·Evgeniy V DubrovinNikolay V Volozhantsev
Apr 16, 2014·Proceedings of the National Academy of Sciences of the United States of America·Byron E PedlerFarooq Azam
Aug 5, 2015·The ISME Journal·Nastassia V PatinPaul R Jensen
Dec 12, 2012·Environmental Microbiology·A K SteenberghH J Laanbroek
Sep 16, 2016·Frontiers in Microbiology·Stefan BraunBente Aa Lomstein
Nov 19, 2017·Applied and Environmental Microbiology·Qiang ZhengNianzhi Jiao
May 12, 2019·Applied and Environmental Microbiology·Alexander KhachikyanMarcel M M Kuypers
Jul 22, 2019·Applied and Environmental Microbiology·Tanja FischerJens Harder
May 16, 2021·Applied and Environmental Microbiology·Dana E MichelsAlexis Pasulka

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