I have images of polished mineral surfaces that have been colonized by numerous types (shapes, sizes, structures) of microbes. I am have used fluorescence microscopy to image these colonized surfaces, with acridine orange as my dye.
I have found filamentous bacteria or fungal hyphae on my surfaces, which have multiple fluorescing nodes which could be individual bacterial cells in the filament or multiple nuclei in the hyphae.
My question this: If I do cell counts to quantify the level of colonization, would you count each fluorescing node in the filament, or consider the whole filament as one cell for the purpose of counting?
Any advice/citations of where people have discussed this in their methods would be appreciated!
Hi Toby, I am absolutely frustated. I spent about 45 Minutes answering - and then the system crashed without saving. Would you mind talking about that on phone? I am 95% sure the long guys are fungal hyphae - not easy to make reliable counting with hyphal networks. Depends on what you need to count really. Scaling would have been nice.
What do you think the big round spots are?
Let phone or skype on it, tomorrow or Thursday? Feel free to send me a phone number,
best wishes, Annette
Hi Annette, That's my bad on the scaling - the magnification of the image is 1000x with the image being roughly 85um x 65um. I can send you images with scale bars if you like.
If by the larger spots in the filaments I have no idea, but there are a range of cell sizes in the image - I think some of them are likely eukaryotic with what look like nuclei.
Skype would be better, I will send you an email.
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