guest ::
| Home > Publications database > Enumeration of soil bacteria with the green fluorescent nucleic acid dye Sytox green in the presence of soil particles |
| Home > Publications database > Enumeration of soil bacteria with the green fluorescent nucleic acid dye Sytox green in the presence of soil particles |
| Journal Article | PreJuSER-40216 |
; ; ; ;
2004
Elsevier
New York, NY
This record in other databases: 
Please use a persistent id in citations: doi:10.1016/j.mimet.2004.07.004
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.
Keyword(s): Aluminum Silicates (MeSH) ; Bacillus subtilis: isolation & purification (MeSH) ; Bacillus subtilis: metabolism (MeSH) ; Cell Membrane: metabolism (MeSH) ; Fluorescent Dyes: chemistry (MeSH) ; Fluorescent Dyes: metabolism (MeSH) ; Microscopy, Fluorescence: methods (MeSH) ; Organic Chemicals (MeSH) ; Pseudomonas: isolation & purification (MeSH) ; Pseudomonas: metabolism (MeSH) ; Ralstonia: isolation & purification (MeSH) ; Ralstonia: metabolism (MeSH) ; Soil Microbiology (MeSH) ; Spectrophotometry, Ultraviolet (MeSH) ; Aluminum Silicates ; Fluorescent Dyes ; Organic Chemicals ; SYTOX Green ; clay ; J ; Sytox green (auto) ; total counts (auto) ; digital image analysis (auto)
|
The record appears in these collections: |