My best Amphipleura pellucida result - so far

Begonnen von Beatsy, März 24, 2023, 17:24:18 NACHMITTAGS

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Beatsy

Hello everyone.

I am building a microscope setup to photograph diatoms using 365nm UV illumination (to increase resolution). The project is not complete but I reached a milestone today by capturing a picture of Amphipleura pellucida with the pores nicely resolved. I'm very pleased with it and even though there are obvious flaws with uneven background and halos (being actively resolved) I'm itching to show this result "so far". So here it is.

The 1st image is a crop from the 2nd one to show the detail more clearly. It was enlarged 2x in Topaz Gigapixel AI. The 2nd image is a 100% pixel-for-pixel crop from the captured frame. Single picture (4-shot pixel shift), no stacking or sharpening - only contrast stretching.

Gear used:
Olympus BX61 fluorescence microscope. Heine condenser with light from a Convoy 365nm UV torch reflected into it. Olympus UPlanSApo 100/1.4 objective. The Heine and objective were both oiled to the slide. Camera is a full-spectrum-conversion, Sony A7riv connected to the existing C-Mount adapter with direct projection onto the sensor.

I used DPX (DePeX) as the mountant because it doesn't fluoresce under UV. It's only RI 1.52 but that didn't seem to matter with (effectively) circular oblique light from the Heine. Contrast was good enough. However, I have a UV pass (visible block) filter on the way which will solve the fluorescence problem (by blocking it), so zrax (RI 1.7+) will soon be useable on this system too.

Thanks for looking.
Knowledge is cheap. Experience is not.

Siegfried

Hello Steve
A very challenging project and a convincing first result. Please keep reporting on your progress.
   Greetings from Siegfried

Hallo Steve
ein sehr anspruchsvolles Projekt und ein überzeugendes erstes Ergebnis. Bitte berichte weiter über deine Fortschritte.
   Grüße von Siegfried

Beatsy

Thank you Seigfried.

As it happens, the UV-pass filter arrived today and I had to try it out right away. I was able to photograph another A. pellucida, but this time mounted in zrax. It fluoresced, but only UV light got to the sensor. This yielded more contrast as expected. I grabbed one 16-shot pixel shifted image (4x more megapixels over the same field of view) and cropped in really tight to show the detail captured. No enlargement done on this one, it's just a 100% pixel-for-pixel crop.


Knowledge is cheap. Experience is not.

anne

Hi Steve,
I would not have expected the influence of the RI to be so high in the UV range. However, it is amazing to see the pores so clearly even at a low RI with DPX.
Looking forward for amazing images!
best
anne

(And I learned a new thing, "pixel shift", my Olympus can do this!)