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Fluorescence in Situ Hybridization (FISH)

Fluorescence in Situ Hybridization (FISH) is a microscopic procedure that allows for the visualization of specific genes using a fluorescent marker and viewing the samples using an epifluorescent microscope. FISH is used in a variety of different fields of science, including genetics, microbiology, cell biology, embryology, just to name a few. In microbiology, FISH is used to identify specific groups of bacteria in environmental samples, such as wastewater, wetlands, etc. Oligonucleotide probes, short pieces of DNA/RNA, are designed to attach to specific genes. In the case of these samples, we used FISH to identify Nitrifying bacteria in the system. We used probes designed to attach to Nitrosomonas species and Nitrobacter species (the most common Nitrifiers).

The photos below represent the wastewater samples with the probes NIT (targets Nitrobacter species) and NSO probe (targets Nitrosomonas species), as well as the sample spiked with a Nitrifier product, and straight Nitrifier product. Based on the FISH probes, there does not appear to be many nitrifying bacteria in the system compared to the spiked sample and the control sample.

FISH probe photos:

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200X. Fluorescent Image. Effuent. NSO probe attached to Nitrosomonas species.
200X Fluorescent image. Effluent. NIT probe attached to Nitrobacter species.
200X Fluorescent image. Effluent. NIT probe attached to Nitrobacter species.

 

200X. Fluorescent image. Effluent sample with probe NSO spiked with Nitrifiers.
200X. Fluorescent image. Effluent sample with probe NSO spiked with Nitrifiers.
200X Fluorescent image. Effluent sample with probe NIT spiked with Nitrifiers.
200X Fluorescent image. Effluent sample with probe NIT spiked with Nitrifiers.

 

200X Fluorescent image. Nitrifier sample only with NSO probe.
200X Fluorescent image. Nitrifier sample only with NSO probe.
200X Fluorescent image. Nitrifier sample only with NIT probe.
200X Fluorescent image. Nitrifier sample only with NIT probe.