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@ARTICLE{Klauth:40216,
author = {Klauth, P. and Wilhelm, R. and Klumpp, E. and Poschen, L.
and Groeneweg, J.},
title = {{E}numeration of soil bacteria with the green fluorescent
nucleic acid dye {S}ytox green in the presence of soil
particles},
journal = {Journal of microbiological methods},
volume = {59},
issn = {0167-7012},
address = {New York, NY},
publisher = {Elsevier},
reportid = {PreJuSER-40216},
pages = {189 - 198},
year = {2004},
note = {Record converted from VDB: 12.11.2012},
abstract = {Total counts in soils are usually determined using
fluorescent dyes, such as DAPI or Sybr green, due to
fluorescence enhancement if they are bound to nucleic acids.
Unfortunately, these commonly used dyes stain soil particles
as well. Therefore, besides fluorescence enhancement,
sufficient spectral differentiation is also required. We
present a new procedure that overcomes the problems of
visualising bacteria on surfaces in soil and avoids the
separation of soil particles to a large extent. Spectral
differentiation between bacteria and soil matrix is achieved
by using Sytox green and a suboptimal excitation wavelength.
Bacteria exhibit a bright green fluorescence, while soil
particles fluoresce blue or red. Slight homogenisation and
sedimentation of the sand and coarse silt that were too big
for microscopic investigations were the only separation
steps required. We compared the proposed Sytox green
staining with Sybr green staining. The recovery of Sybr
green-stained cells amounted to $38\%,$ whereas in samples
stained by Sytox green $81\%$ of the spiked cells were
counted. Sytox green can also be combined with fluorescence
in situ hybridisation (FISH) using deep red dyes such as
Cy5.},
keywords = {Aluminum Silicates / Bacillus subtilis: isolation $\&$
purification / Bacillus subtilis: metabolism / Cell
Membrane: metabolism / Fluorescent Dyes: chemistry /
Fluorescent Dyes: metabolism / Microscopy, Fluorescence:
methods / Organic Chemicals / Pseudomonas: isolation $\&$
purification / Pseudomonas: metabolism / Ralstonia:
isolation $\&$ purification / Ralstonia: metabolism / Soil
Microbiology / Spectrophotometry, Ultraviolet / Aluminum
Silicates (NLM Chemicals) / Fluorescent Dyes (NLM Chemicals)
/ Organic Chemicals (NLM Chemicals) / SYTOX Green (NLM
Chemicals) / clay (NLM Chemicals) / J (WoSType)},
cin = {ICG-IV},
ddc = {570},
cid = {I:(DE-Juel1)VDB50},
pnm = {Chemie und Dynamik der Geo-Biosphäre},
pid = {G:(DE-Juel1)FUEK257},
shelfmark = {Biochemical Research Methods / Microbiology},
typ = {PUB:(DE-HGF)16},
pubmed = {pmid:15369855},
UT = {WOS:000224363000005},
doi = {10.1016/j.mimet.2004.07.004},
url = {https://juser.fz-juelich.de/record/40216},
}