Ant Eyes

Images made through a microscope. All subject types.

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RogelioMoreno
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Ant Eyes

Post by RogelioMoreno »

I was testing a new 20X objective and my ping pong ball diffuser.

Nikon 10/0.3 WI 160/- (without the WI tipping), direct projection (I removed the trinocular head and put the camera on the Nikon Labophot stand), ping pong ball diffuser, single FO illuminator, 273 frames with 4um steps:
Image


L Plan 20/0.4 00/0 with Vivitar 200mm M42 acting as tube lens (I removed the trinocular head and put the camera + Vivitar 200mm + M42 spacers on the Nikon Labophot stand), ping pong ball diffuser, single FO illuminator, 98 frames with 2um steps:
Image

Comments are welcome.

Rogelio

elf
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Joined: Sun Nov 18, 2007 12:10 pm

Post by elf »

The results are quite good. How much post processing have you done to these?

The step distance seems kind of small. The 10X's DOF should be around 15um. Do you get any focus banding if you stack every other frame or every third frame?

RogelioMoreno
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Joined: Fri Nov 20, 2009 11:24 am
Location: Panama

Post by RogelioMoreno »

Elf,

I did contrast adjustment and sharpener. I think that in the first image I should had done some brightness adjustment.

The formula that I am using to calcule DOF says 5.7um DOF for a 0.3 NA. I have to try to stack without some frames.

Rogelio

RogelioMoreno
Posts: 2979
Joined: Fri Nov 20, 2009 11:24 am
Location: Panama

Post by RogelioMoreno »

The previous images were stacked with HF, here are the version stacked with ZS:

Image


Image


Rogelio

rjlittlefield
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Post by rjlittlefield »

Rogelio,

Thanks for posting the comparison. I assume these are ZS PMax. I see the usual tradeoffs: halos are gone around edges but there is more bloom around bright reflections; structure is retained in out-of-focus regions, but there is more pixel noise especially in uniform background.

I also see irregular dust trails and some rotated streaking on edges, especially in the first image. Those artifacts usually mean that the ZS alignment algorithm has gotten confused by how things look as they go severely OOF.

For this sort of high magnification work, it is usually best to go into Options > Preferences > Alignment and turn off Scale and Rotation. I believe your focus mechanism moves straight up and down, so probably you can also turn off Shift X and Shift Y. Doing those things will eliminate the dust trails and streaking.

I am thinking the ZS user interface could use some improvement. Right now there is one set of default settings. Those work well at lower magnifications, but as we see here they can give odd results at higher mags, and it's never obvious to new users what is going on. I will need to think about this...thanks for the illustrations.

--Rik

RogelioMoreno
Posts: 2979
Joined: Fri Nov 20, 2009 11:24 am
Location: Panama

Post by RogelioMoreno »

Rik,

Thank you for the info, I will try that.

Rogelio

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