Atomic Force Microscopy - Laser Scanning Confocal Microscopy imaging protocol for copepod fecal pellets
Malfatti, F.; Cnudde, C.; De Troch, M. (2013). Atomic Force Microscopy - Laser Scanning Confocal Microscopy imaging protocol for copepod fecal pellets, in: Cnudde, C. Trophic ecology of intertidal harpacticoid copepods, with emphasis on their interactions with bacteria = Trofische ecologie van intertidale harpacticoide copepoden, met de nadruk op hun interacties met bacteriën. pp. 127-134
In: Cnudde, C. (2013). Trophic ecology of intertidal harpacticoid copepods, with emphasis on their interactions with bacteria = Trofische ecologie van intertidale harpacticoide copepoden, met de nadruk op hun interacties met bacteriën. PhD Thesis. Ghent University (UGent): Gent. ISBN 9789090278285. 209 pp., meer
Within the marine carbon and nitrogen biogeochemical cycles, fecal pellets produced by pelagic and benthic copepods are important microbial activity hotspots as particle substrate for bacteria colonization and as sources of particulate and dissolved organic C and N. We developed a protocol combining Atomic Force Microscopy (AFM) and Laser Scanning Confocal Microscopy (LSCM) to study the peritrophic membrane structure and associated bacteria of fecal pellets produced by a benthic copepod, Paramphiascella fulvofasciata. AFM imaging revealed a fibrillar network structure of the peritrophic membrane, 0.7-5.9 nm thick similar to marine polysaccharides and a-chitin. Bacterial cell volume range was 0.006-0.117 µm3 in liquid. LSCM imaging showed a 3D-heterogeneous microenvironment. This protocol would allow high-resolution interrogation of structural changes and bacterial dynamics within the copepod fecal pellets and other heterogeneous particles such marine snow under environmental conditions.
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