At the moment I am working with Fluorescence in Situ Hybridization to study microbial communities in activated sludge
Samples which I've recently studied posses a strong tendence to autofluorescence. I've tried to deal with this problem by making some procedural modifications or reducing the amount of some extracellular substances.
But I am wondering if there are more ways (maybe more effective) to minimalize the autofluorenscence of activated sludge.
That's a tough one. I'm not a "sludge" expert, but unlike with cells in culture or tissue samples, you can't easily do a sodium borohydride treatment. So I think your options would be: 1) choose dyes that are outside the autofluorescent spectrum of the sample, 2) image with one wavelength left open (specific label in green, and an "empty" channel in red) then subtract one image from the other using imaging software to artificially subtract out the autofluorescent signal, or 3) photobleach the sample with very strong UV light prior to labeling in the hope that the autofluorescent compounds will fade and not come back.
Hello, Anna,
I do not have enough information on your work to fully answer this question. However, I myself observed a strange and transient phenomenon in the behavior of settling activated sludge. The hypothesis of a transient emission of light is certainly not excluded.
I would advise reading my articles:
Thierie J. (2012)
Near-infrared dynamic measurements of activated sludge highlight the possibility of the local modification of free water properties.
J. Near Infrared Spectrosc. 20:415 - 418
Thierie J. (2013)
NIR observation of activated sludge decantation indicates correlation with the effluent suspended solids of four different wastewater treatment plant situations.
Microbial & Biochemical Technology. 5:130-135.
(published on Researchgate)
The other hypothesis is light emission by flocs ...
I continue to work on this phenomenon and I would be happy that we can continue to exchange if our concerns are close.
Best, Jacques
Hello,
Thank you so much for all responses. I am certain that all of the information may be helpful.
Jacques, I am going to read your articles. Thank you again.
In general, I think that this topic is interesting.
Despite I take the samples from one reactor, one sample possess stronger tendency to autofluorescence and one not (I think there may be a few reasons).
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