Yellow Jacket

Post your images made through a compound microscope or made with a stereo/dissecting microscope in this gallery. Images may be of any subject natural or unnatural, living or non-living.

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Frez
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Yellow Jacket

Post by Frez »

I worked hard trying to get a good stack with a 4x objective in both CZ and HF. It just wouldn't come out right. Any suggestions or tips...

Thanks
Frez

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piotr
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Post by piotr »

For me they already look amazing! What unearthly pictures!!! :shock:
Great work!
Piotr

rjlittlefield
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Post by rjlittlefield »

Frez,

In the upper one, I see what looks like radial streaking. Is that the problem you see? Does it show in the individual images? If so, then you've got some sort of optical problem. If not (which I suspect), then you've got some sort of auto-adjustment problem -- the stacking software is not properly registering the various frames. Odd that it would happen with both CZ and HF. What settings are you using?

--Rik

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Frez
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Post by Frez »

Thanks Piotr. I bought a new HBO lamp and it made a big difference. The old one was starting to blacken on the inside and was casting shawdows on the image. There's an eBay seller that offers them at $75 USD. The cheapest found on the web was $110.

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Frez
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Post by Frez »

Rik, The problems are within the left eye and the upper portion of the right one. At first I thought the pattern of the eyes was causing problems with the software, but the lower portion of the right eye looks ok. I tried a stack of 49 and another of 103 with new frames. Both had the same result. Reviewing the frames is difficult because they are all dark. The final image needs a hefty levels adjustment in PS after removing noise in Neat Image. This new bulb is brighter and allows the use of an Olympus FK 3.3x photo eyepiece with a 10x objective. There doesn't seem to be enough light to work with the 4x obj though. Maybe tonight I'll try the 4x with a .36x relay instead. It vignettes, but gathers all of the available light.

Frez

rjlittlefield
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Post by rjlittlefield »

Frez,

I think we're looking at the same effect.

This still feels like a problem getting the images properly aligned.

You might investigate a couple of ways. 1) Run the same stack without auto-adjustment and see if it changes. 2) Shoot a white light stack (I presume you've got plenty of light then), check the individual frames, run it with and without auto-adjustment and see if either composite looks the same as the fluorescent stack.

It could be that your problem is all due to image brightness and noise. Sometimes image processing algorithms have thresholds programmed into them to ignore small differences between pixel values, on the assumption that they're meaningless noise. I haven't studied the auto-adjustment parts of either HF or CZ, so I don't know if they have noise thresholds.

One possibility would be to apply the level adjust and possibly Neat Image to the input frames, before sending them to the stacking software. In Photoshop, File | Automate | Batch... would avoid having to touch each image individually.

Also, in CZ there is some way to specify the scale change between images. You might shoot a white stack with the same step size, see what CZ infers for scale change in that case, then force the same value for the fluorescent stack. This is getting down to hacking, though. It would be a lot better to get the auto-adjustment working.

--Rik

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