Fossil Diatoms of Kamyshlov Russia
Fossil Diatoms of Kamyshlov Russia
Another gifted slide, These are fossil diatoms from Kamyshlov, Ruzzia
All are stacks 4 to 7 images. Don't know why these proved to be so challenging, but I am not completely happy with the results. Any pointers are welcome.
AO 10, Ortho Illuminator, 100x Iris Oil Plan Achro, Toric Darkfield Condenser, Canon 70D
All are stacks 4 to 7 images. Don't know why these proved to be so challenging, but I am not completely happy with the results. Any pointers are welcome.
AO 10, Ortho Illuminator, 100x Iris Oil Plan Achro, Toric Darkfield Condenser, Canon 70D
Re: Fossil Diatoms of Kamyshlov Russia
Rod,
They look very good to me. With your darkfield rig, I doubt you can improve them much, except for using green filter or modern plan apos.
The 1st diatom may be quite thick and three dimensional (and naturally difficult for darkfield at high NA/magnification). It may image better, with much less halos, in brightfield or oblique.
I remember Steve Beats (or Charles Krebs?) did some diatoms stacks in brightfield or oblique, then digitally inverted the colors with softwares, generating some clean-looking "digital darkfield" images. Maybe give that a try? I remember forum member kit1980 did some "digital darkfield" on fungus images as well.
They look very good to me. With your darkfield rig, I doubt you can improve them much, except for using green filter or modern plan apos.
The 1st diatom may be quite thick and three dimensional (and naturally difficult for darkfield at high NA/magnification). It may image better, with much less halos, in brightfield or oblique.
I remember Steve Beats (or Charles Krebs?) did some diatoms stacks in brightfield or oblique, then digitally inverted the colors with softwares, generating some clean-looking "digital darkfield" images. Maybe give that a try? I remember forum member kit1980 did some "digital darkfield" on fungus images as well.
Re: Fossil Diatoms of Kamyshlov Russia
Thanks zz,
Your point about realistic expectations of darkfield is a good one.
I do think I could do a better job with incrementing the focus steps when shooting stacks and it would have a positive impact. I need to be more precise and use the scale on the focuser knob instead of winging it.
Thanks for responding
Rod
Your point about realistic expectations of darkfield is a good one.
I do think I could do a better job with incrementing the focus steps when shooting stacks and it would have a positive impact. I need to be more precise and use the scale on the focuser knob instead of winging it.
Thanks for responding
Rod
Re: Fossil Diatoms of Kamyshlov Russia
Rod,
You may try half of a division per turn per image. I cannot remember the exact number, though you want to cover each increment of depth of view with 2-3 steps, iirc.
You may try half of a division per turn per image. I cannot remember the exact number, though you want to cover each increment of depth of view with 2-3 steps, iirc.
Re: Fossil Diatoms of Kamyshlov Russia
At first glance I'd have thought that your microscope resolution was exceeding your camera resolution, but after looking at the specs of your camera I'm not sure that's the case. So, I guess I'm not of much assistance.
lorez
lorez
Re: Fossil Diatoms of Kamyshlov Russia
Thanks - now define "increment of depth". Seems like that could be one of those infinite type things. Not sure I am up for a 1500 image stack ;^).zzffnn wrote:Rod,
You may try half of a division per turn per image. I cannot remember the exact number, though you want to cover each increment of depth of view with 2-3 steps, iirc.
But I will try again.
Thank zz.
Rod
Re: Fossil Diatoms of Kamyshlov Russia
Yes, I think I am covered on the camera front, that leaves only operator error as the answer, and I am quite willing to believe that solution.lorez wrote:At first glance I'd have thought that your microscope resolution was exceeding your camera resolution, but after looking at the specs of your camera I'm not sure that's the case. So, I guess I'm not of much assistance.
lorez
Thanks for thinking on it.
Rod
Re: Fossil Diatoms of Kamyshlov Russia
Rod,
I am not positive about it. If you think more accurate stepping would help there - I remember Rik said if depth of field (DoF) is 1 micron, then each stacking step should ideally be 0.3-0.5 microns. But realistically, with a microscope focus, one can only reliably step at 0.5-1 micron, no less. Your iris 100x, even when stopped down by iris, should still have DoF of around 0.5 microns:
https://www.microscopyu.com/microscopy- ... h-of-focus
Also, Rik's ideal stepping depth is more for hairy macro subject under reflected light. Your diatoms in high NA darkfield is very different than that.
Another thing to try is to stack some sub-layers in Pmax (say photos 1-10 as layer #1, 11-20 as layer #2), then combine all layers together (in Dmax or Pmax). That can probably reduce halo/noise/image contamination from layers above and below. Remember Pmax will accumulate noise, if you do one huge stack, instead of sub layer stacks. I think Steve Beats did sublayer stacks sometimes. It is going to be a lot of work though.
I am not positive about it. If you think more accurate stepping would help there - I remember Rik said if depth of field (DoF) is 1 micron, then each stacking step should ideally be 0.3-0.5 microns. But realistically, with a microscope focus, one can only reliably step at 0.5-1 micron, no less. Your iris 100x, even when stopped down by iris, should still have DoF of around 0.5 microns:
https://www.microscopyu.com/microscopy- ... h-of-focus
Also, Rik's ideal stepping depth is more for hairy macro subject under reflected light. Your diatoms in high NA darkfield is very different than that.
Another thing to try is to stack some sub-layers in Pmax (say photos 1-10 as layer #1, 11-20 as layer #2), then combine all layers together (in Dmax or Pmax). That can probably reduce halo/noise/image contamination from layers above and below. Remember Pmax will accumulate noise, if you do one huge stack, instead of sub layer stacks. I think Steve Beats did sublayer stacks sometimes. It is going to be a lot of work though.
Re: Fossil Diatoms of Kamyshlov Russia
Thanks zzzzffnn wrote:Rod,
I am not positive about it. If you think more accurate stepping would help there - I remember Rik said if depth of field (DoF) is 1 micron, then each stacking step should ideally be 0.3-0.5 microns. But realistically, with a microscope focus, one can only reliably step at 0.5-1 micron, no less. Your iris 100x, even when stopped down by iris, should still have DoF of around 0.5 microns:
https://www.microscopyu.com/microscopy- ... h-of-focus
Also, Rik's ideal stepping depth is more for hairy macro subject under reflected light. Your diatoms in high NA darkfield is very different than that.
Another thing to try is to stack some sub-layers in Pmax (say photos 1-10 as layer #1, 11-20 as layer #2), then combine all layers together (in Dmax or Pmax). That can probably reduce halo/noise/image contamination from layers above and below. Remember Pmax will accumulate noise, if you do one huge stack, instead of sub layer stacks. I think Steve Beats did sublayer stacks sometimes. It is going to be a lot of work though.
These are relatively small stacks, but of course should I reduce my increments, the count could climb. I'll see how it goes. Thanks for the thoughts.
Rod
Re: Fossil Diatoms of Kamyshlov Russia
I have to say that I like them very much: they are certainly far better than anything I can do.