TY  - JOUR
AU  - Klauth, P.
AU  - Wilhelm, R.
AU  - Klumpp, E.
AU  - Poschen, L.
AU  - Groeneweg, J.
TI  - Enumeration of soil bacteria with the green fluorescent nucleic acid dye Sytox green in the presence of soil particles
JO  - Journal of microbiological methods
VL  - 59
SN  - 0167-7012
CY  - New York, NY
PB  - Elsevier
M1  - PreJuSER-40216
SP  - 189 - 198
PY  - 2004
N1  - Record converted from VDB: 12.11.2012
AB  - 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.
KW  - Aluminum Silicates
KW  - Bacillus subtilis: isolation & purification
KW  - Bacillus subtilis: metabolism
KW  - Cell Membrane: metabolism
KW  - Fluorescent Dyes: chemistry
KW  - Fluorescent Dyes: metabolism
KW  - Microscopy, Fluorescence: methods
KW  - Organic Chemicals
KW  - Pseudomonas: isolation & purification
KW  - Pseudomonas: metabolism
KW  - Ralstonia: isolation & purification
KW  - Ralstonia: metabolism
KW  - Soil Microbiology
KW  - Spectrophotometry, Ultraviolet
KW  - Aluminum Silicates (NLM Chemicals)
KW  - Fluorescent Dyes (NLM Chemicals)
KW  - Organic Chemicals (NLM Chemicals)
KW  - SYTOX Green (NLM Chemicals)
KW  - clay (NLM Chemicals)
KW  - J (WoSType)
LB  - PUB:(DE-HGF)16
C6  - pmid:15369855
UR  - <Go to ISI:>//WOS:000224363000005
DO  - DOI:10.1016/j.mimet.2004.07.004
UR  - https://juser.fz-juelich.de/record/40216
ER  -