Hey fellow researchers,
We seem to be obtaining low resolution images under all magnifications for actinobacteria on transient slides prepared from liquid and solid cultures. Can you suggest any ways to improve the quality of the images, especially in the preparation of slides and toying with the settings of the optical microscope ?
Your inputs would be greatly appreciated.
Best Wishes
Dilantha
Dilantha, are you using a digital or an analogue camera? That means, do you get your images directly in files on a computer or on film?
What kind of staining do you apply?
How does your microscope perform if you examine a completely different sample (something totally unrelated to bacteria)?
Please provide some more information.
Hi,
It is digital and I checked for the image quality by looking at some Nostoc and the resolution of the cyanobacterium was good - there was clear cell types and I played around with the enhancements and the color tools and I was able to get a clear image
As for the actinobacteria hyphae, there were a lot of filaments but the resolution was low and since there were many planes I suppose, part of the viewing area was clear and others were not.
I'm not very experienced with microscopy since I'm a molecular biologist/biochemist but I'm trying to get some clear images to contrast some related actinobacteria.
The microscope model is Optika B-350.
The stains used were lactophenol cotton blue, safranine, and crystal violet.
Thank you for your inputs so far .......
Best Wishes
Dilantha
I guess you are right with the focal plane. Your sample may be too thick. Can you make some dilutions or separate very small parts of the hyphae under a stereomicroscope?
Smearing of your sample with a cover glass may also help to distribute the hyphae on the glass slide. The image below illustrates the method. The example is for blood samples, but it also works for fungal hyphae. You only need some small areas where there are not so many cells.
Good luck.
http://www.funsci.com/fun3_en/usph/usph_18.gif
You may also try extended depth of field. Basiacally you will make several images at different focus planes and then let computer to make one sharp one. Fiji has plugin for it.
Hi,
Thank you both for your valuable suggestions. I will try to use these techniques and see whether the focus is stronger and images are of high resolution.
Best,
Dilantha
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