Fossil Diatoms of Kamyshlov Russia

Here you can post pictures and videos to show others.
Post Reply
Message
Author
User avatar
rnabholz
Posts: 3086
Joined: Thu Jan 08, 2015 10:11 pm
Location: Iowa USA
Contact:

Fossil Diatoms of Kamyshlov Russia

#1 Post by rnabholz » Fri Apr 28, 2017 1:07 am

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
61µm
61µm
Kamyshlov Russia Diatom.jpg (99.05 KiB) Viewed 5323 times
38µm
38µm
Kamyshlov Russia Centric 1.jpg (87.17 KiB) Viewed 5323 times
46µm
46µm
Kamyshlov Russia Centric 2.jpg (104.17 KiB) Viewed 5323 times

User avatar
zzffnn
Posts: 3200
Joined: Sat Jun 20, 2015 3:57 am
Location: Houston, Texas, USA
Contact:

Re: Fossil Diatoms of Kamyshlov Russia

#2 Post by zzffnn » Fri Apr 28, 2017 4:10 am

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.

User avatar
rnabholz
Posts: 3086
Joined: Thu Jan 08, 2015 10:11 pm
Location: Iowa USA
Contact:

Re: Fossil Diatoms of Kamyshlov Russia

#3 Post by rnabholz » Fri Apr 28, 2017 8:54 pm

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

User avatar
zzffnn
Posts: 3200
Joined: Sat Jun 20, 2015 3:57 am
Location: Houston, Texas, USA
Contact:

Re: Fossil Diatoms of Kamyshlov Russia

#4 Post by zzffnn » Fri Apr 28, 2017 10:28 pm

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.

User avatar
lorez
Posts: 735
Joined: Wed Dec 17, 2014 1:48 am

Re: Fossil Diatoms of Kamyshlov Russia

#5 Post by lorez » Fri Apr 28, 2017 11:18 pm

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

User avatar
rnabholz
Posts: 3086
Joined: Thu Jan 08, 2015 10:11 pm
Location: Iowa USA
Contact:

Re: Fossil Diatoms of Kamyshlov Russia

#6 Post by rnabholz » Sat Apr 29, 2017 12:56 am

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.
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 ;^).

But I will try again.

Thank zz.

Rod

User avatar
rnabholz
Posts: 3086
Joined: Thu Jan 08, 2015 10:11 pm
Location: Iowa USA
Contact:

Re: Fossil Diatoms of Kamyshlov Russia

#7 Post by rnabholz » Sat Apr 29, 2017 12:58 am

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
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.

Thanks for thinking on it.

Rod

User avatar
zzffnn
Posts: 3200
Joined: Sat Jun 20, 2015 3:57 am
Location: Houston, Texas, USA
Contact:

Re: Fossil Diatoms of Kamyshlov Russia

#8 Post by zzffnn » Sat Apr 29, 2017 2:47 am

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.

User avatar
rnabholz
Posts: 3086
Joined: Thu Jan 08, 2015 10:11 pm
Location: Iowa USA
Contact:

Re: Fossil Diatoms of Kamyshlov Russia

#9 Post by rnabholz » Sat Apr 29, 2017 9:51 pm

zzffnn 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.
Thanks zz

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

User avatar
gekko
Posts: 4701
Joined: Sun Oct 12, 2014 7:38 am
Location: Durham, NC, USA.

Re: Fossil Diatoms of Kamyshlov Russia

#10 Post by gekko » Thu May 04, 2017 3:07 pm

I have to say that I like them very much: they are certainly far better than anything I can do.

Post Reply