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MBD0049

Sigma-Aldrich

Escherichia coli FISH probe -ATTO488

Probe for fluorescence in situ hybridization (FISH), 20µM in water

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Quality Level

200

technique(s)

FISH: suitable

fluorescence

λex 504 nm; λem 521 nm (ATTO488)

shipped in

dry ice

storage temp.

−20°C

General description

Fluorescent In Situ Hybridization technique (FISH) is based on the hybridization of fluorescent labeled oligonucleotide probe to a specific complementary DNA or RNA sequence in whole and intact cells.1 Microbial FISH allows the visualization, identification and isolation of bacteria due to recognition of ribosomal RNA also in unculturable samples.2

The FISH technique was successfully used to identify E.coli with the probe in various samples such as pure culture (as described in the figure legends and11,13), large and small intestines samples14-16, fecal samples17-21, colonic biopsies18, urine samples, bladder, and kidney sections embedded in paraffin22 and in E.coli biofilm23. It is strongly recommended to include positive and negative controls in FISH assays to ensure specific binding of the probe of interest and appropriate protocol conditions. We offer positive (MBD0032/33) and negative control (MBD0034/35) probes, that accompany the specific probe of interest.

Escherichia coli is a gram negative, facultative aerobic, rod-shaped coliform bacterium. E. coli colonizes the infant gut within hours of birth and establishes itself as the most abundant facultative anaerobe of the human intestinal microflora for the remainder of life, equipped with the abilities to grow in the ever-changing environment in the gut and cope with the mammalian host interaction.8,9 Nevertheless, E. coli can survive in many different ecological habitats, including abiotic environments, and is considered a highly versatile species. Known habitats of E. coli include soil, water, sediment, and food. Some strains of E. coli have evolved and adapted to a pathogenic lifestyle and can cause different disease pathologies.10

Escherichia coli probe specifically recognizes Escherichia coli cells. Yet some reports describe the recognition of other bacteria with this probe, such as, Shigella boydii, Citrobacter davisae, Citrobacter lapagei, Citrobacter neteri11 and Klebsiella pneumoniae12.

Application

Probe for fluorescence in situ hybridization (FISH), recognizes Escherichia coli cells

Features and Benefits

  • Visualize, identify and isolate Escherichia coli cells.
  • Observe native E. coli cell populations in diverse microbiome environments.
  • Specific, sensitive and robust identification of E. coli in bacterial mixed population.
  • Specific, sensitive and robust identification even when E. coli is in low abundance in the sample.
  • FISH can complete PCR based detection methods by avoiding contaminant bacteria detection.
  • Provides information on E. coli morphology and allows to study biofilm architecture.
  • Identify E. coli in clinical samples such as, urine samples, bladder and kidney sections (formalin-fixed paraffin-embedded (FFPE) samples), fecal samples and colon tissue.
  • The ability to detect E. coli in its natural habitat is an essential tool for studying host-microbiome interaction.

Storage Class Code

12 - Non Combustible Liquids

WGK

WGK 1

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable

Regulatory Information

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P V Kirjavainen et al.
Gut, 51(1), 51-55 (2002-06-22)
Recent data have outlined a relationship between the composition of the intestinal microflora and allergic inflammation, and demonstrated the competence of probiotics in downregulation of such inflammation. Our aims were to characterise the relationship between gut microbes and the extent
Rafael Gustavo Hermes et al.
The British journal of nutrition, 109(6), 1001-1012 (2012-08-02)
Casein glycomacropeptide (CGMP), a glycoprotein originating during cheese manufacture, has shown promising effects by promoting the growth of some beneficial bacteria in vitro, although its activity has not been well explored. The present study was designed to evaluate the effects
D Grimm et al.
Applied and environmental microbiology, 64(7), 2686-2690 (1998-07-02)
Based on comparative sequence analysis, we have designed an oligonucleotide probe complementary to a region of 16S rRNA of Legionella pneumophila which allows the differentiation of L. pneumophila from other Legionella species without cultivation. The specificity of the new probe
Stuart C Smith et al.
European journal of nutrition, 45(6), 335-341 (2006-06-10)
Changes in the composition of gastrointestinal microbiota by dietary interventions using pro- and prebiotics provide opportunity for improving health and preventing disease. However, the capacity of lupin kernel fiber (LKFibre), a novel legume-derived food ingredient, to act as a prebiotic
L K Poulsen et al.
Infection and immunity, 62(11), 5191-5194 (1994-11-01)
Fluorescent oligonucleotide probes targeting rRNA were used to develop an in situ hybridization technique by which the spatial distribution of Escherichia coli in the large intestines of streptomycin-treated mice was determined. Single E. coli cells were identified in thin frozen
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