MicrobeHunter.com Microscopy Forum

You say that the microscope is set up for phase contrast. Are these all phase images?
In the photo of your setup it looks as though the condenser is set to brightfield?

Some are in phase, however most are brightfield. As for the RAW file i would love to send one once i get home. I’ll use something more organized such as onion epidermis if this is better than trying to photograph rotifers.

As others have said, the photos are underexposed.
What is preventing you from setting the correct exopsure in Canon Utilities?

Well the issue is just mainly balancing shutter speed and iso, however i adjusted the exposer compensation on the camera just to experiment and it worked, however the pictures are still grainy. I’ll post some soon

So here is some more pictures with some differing settings
The original picture was taken at 0"6 shutter speed and 100 ISO with Kholer illumination and in Bright field.


Next i toyed around in Photo Pos Pro and got this as a result:


I need to do some more fine tuning but i may be on to something(?). One issue i currently have is making my camera par-focal, it just wont do it. Anyways i hope you enjoy and can help me improve even more

It may sound oversimplified but remember that what anyone that takes a photo through a microscope is trying to do is reproduce what they see through the eyepieces.

So there are two distinct stages, setting up the microscope correctly and setting up the camera correctly.

Of the two, setting up the microscope correctly is the most challenging.

Despite what people think, post processing is not part of the first stage but part of the second, and no substitute I might add.

Clearing up a distracting background, correcting colour aberrations or giving viewers that do not have the opportunity to look though our eyepieces

a combined view in one image by stacking, will never make up for an uneven microscope setup.


Remember also that empty magnification applies to microscopy, photography and in this digital age to monitors as well.

viewtopic.php?f=25&t=4984&p=45460&hilit#p45460
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Timemaster1212 wrote: ↑

Thu Jun 25, 2020 3:02 am

So here is some more pictures with some differing settings

I'm only seeing the original photo and the edited version.
What objective was used?
Is this the entire view as seen through the eyepiece, or has it been cropped?

Ah i did not consider empty magnification, i will look more into that. And I totally forgot to mention it, but it was an 43x objective with a 10x eyepiece acting as a projecting lens

I have the diatom slide pictured above, but I don't know exactly what to look at. Can someone please tell me what the text/numbers mean below the names of the diatomes (in the white background)?

janvangastel wrote: ↑

Thu Jun 25, 2020 8:12 am

I have the diatom slide pictured above, but I don't know exactly what to look at. Can someone please tell me what the text/numbers mean below the names of the diatomes (in the white background)?

They represent the scale of the ‘graduations on the ruler’
striae are stripes [plural of stria], and the test diatoms are for checking resolution.

It’s rather like a natural version of this:
https://en.m.wikipedia.org/wiki/1951_US ... test_chart

MichaelG.

MichaelG. wrote: ↑

Thu Jun 25, 2020 8:32 am

janvangastel wrote: ↑

Thu Jun 25, 2020 8:12 am

I have the diatom slide pictured above, but I don't know exactly what to look at. Can someone please tell me what the text/numbers mean below the names of the diatomes (in the white background)?

They represent the scale of the ‘graduations on the ruler’
striae are stripes [plural of stria], and the test diatoms are for checking resolution.

It’s rather like a natural version of this:
https://en.m.wikipedia.org/wiki/1951_US ... test_chart

MichaelG.

I have to say that diatoms make for a much prettier ruler ;)

Here is an article on some of the many uses of a Diatom Test Slide: http://www.microscopy-uk.org.uk/mag/ind ... -test.html

Thanks for the clarification. Interesting article.

So i changed my adapter set up, bringing back the tube and attaching the adapter at the bottom of the photo tube. This reduced the empty magnification and, with the blue light filter, made a more pleasant image with some touching up. Is it common for a filter to increase image exposure time dramatically? This is what i can produce now:


edit: this is the same as before, 43x objective with 10x eyepiece for projection

Timemaster1212 wrote: ↑

Thu Jun 25, 2020 6:05 pm

Is it common for a filter to increase image exposure time dramatically?

An 80A photo filter reduces light by about 2 and 2/3 stop. That is the blue filter you would use to correct tungsten to daylight balance, so should be similar to what is in your scope.

Not wishing to flood the active topics, this will be my last post (probably) on this just to show some more experimentation. I read that image stacking can decrease the noise in an image so i attempted this with combine ZP and got the following:


I am not sure if this is just self appreciation, but i am noticing an increase in quality already, thanks to the help of many wonderful community members. I am by no means saying i am close to producing quality images, however i will continue to experiment and improve my methods. It was a great deal of fun "diving into the deep end" so i will see where i end up resurfacing soon.