A Pollen Project. Part 4. The Carnation

Images made through a microscope. All subject types.

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Walter Piorkowski
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A Pollen Project. Part 4. The Carnation

Post by Walter Piorkowski »

A Pollen Project. Part 4. The Carnation
The subject of this fourth post is Dianthus caryophyllus, the Carnation. Its pollen grains are the first circular or maybe even hexagonal that I have seen. At 50 micrometers in diameter, they are medium sized and make a nice composition for the 50x objective.
The pollen grains are somewhat translucent and have a muted glow in the light provided by the incident light Ultropak device. Like Part 3 this made setting the condenser and how to judge the exposure more difficult. What was also interesting about imaging this subject was the different colors and shapes that appeared for the pollen. Some had a yellow hue, others a tan. Still others were green and the most beautiful to my eyes were the blue.
My pollen extraction method for this type of flower cannot discriminate as to what location in the anther structure the pollen grains are coming from. That may have some bearing on the matter. Or possibly the white balance on my camera was going nuts?
I have included an image made with a UO 22x objective and lens condenser.
Walt

Notes:
Leitz Wetzlar Ortholux microscope with ULTROPAK and U.O. 50x, 0.65 NA objective in mirror and lens condenser.
Leitz MIKAS 1/3x micro camera attachment with 1/3x relay lens and 5x eyepiece.
Sony NEX-7, APS-C, 24.3 MP sensor camera.
50x objective magnification on sensor 115x. FOV 0.20 mm. 22x objective magnification 55x on sensor. FOV 0.43 mm
33 and 28 images respectively at 1 micron increments on 50x paired and triple images. Stacked in Zerene software.
Photoshop repair of artifacts.
Attachments
2020-11-28-05.59.03.jpg
2020-11-28-05.53.24-3.jpg
2020-11-28-05.41.09.jpg

iconoclastica
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Re: A Pollen Project. Part 4. The Carnation

Post by iconoclastica »

I think you could make your images even better if you'd run the stack a bit deeper, so the periphery of the pollina will be better defined. I run them to the depth where the deepest spore just has started showing the lower surface, or even somewhat further, for deeper projections sometimes are visible beyond the horizon of the periphery. Visual inspection and reading the micrometer scale of the fine focus will tell where that is.
--- felix filicis ---

rjlittlefield
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Re: A Pollen Project. Part 4. The Carnation

Post by rjlittlefield »

This is an interesting project.

For me, stacking full depth and rendering in stereo with Zerene Stacker would give a lot of insight about the 3D shapes.

Alternatively, stacking just the front and rendering with Helicon's 3D model viewer might work well.

In the images shown here, I have the feeling that the second image shows a fully hydrated spherical form, while the first image shows a dehydrated form that has shrunken around each of the small round features. But that's just from visual appearance and general knowledge of how things dry out, no special knowledge about pollen.

Google image search on Dianthus caryophyllus pollen sem finds a lot of relevant images. Unfortunately many of the papers are located behind paywalls, with rather high prices.

--Rik

Walter Piorkowski
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Re: A Pollen Project. Part 4. The Carnation

Post by Walter Piorkowski »

Thank you, gentlemen, for your input. To be honest iconoclastica, I wish the periphery was more clearly defined. I ran into an issue with the pollen types that were translucent. A large amount of flare is present both visually and photographically, extending around the pollen grain. With the 50x objective anywhere outside the plane of focus there is a zone of fog, for lack of a better word. This of course is normal. But the way the Ultropac illuminates the subject, it seems excessive. I will certainly try your suggestion and hope to see better results.
My subjects are imaged while resting on a piece of deep black glass. I wonder if some of the glare could be a reflection off the surface. I will try getting a focus of the surface of the glass and start up from that point to build my stack. I can always toss what isn’t necessary.
One long held concern is going too deep beyond the point that I believe is in focus. Does adding images that exist before the area of focus is presented muddy the ones that are in focus? Maybe Rik can say whether this idea of mine holds any water.
Rik, I would like to give your rendering in stereo a try once I figure out how to do it. On your idea of dehydration, I believe that you are correct. I saw big changes in the shape of the Royal Sunset Lilly as it dried.
Walt

Beatsy
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Re: A Pollen Project. Part 4. The Carnation

Post by Beatsy »

An interesting set of images. I quite like the partially dehydrated grains for the geometric form they take on.

I think mounting in glycerin is the standard trick to prevent dehydration and shrinkage. Might be worth a try.

Cheers
Beats

iconoclastica
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Re: A Pollen Project. Part 4. The Carnation

Post by iconoclastica »

Beatsy wrote:
Fri Jan 24, 2025 12:42 pm
I think mounting in glycerin is the standard trick to prevent dehydration and shrinkage. Might be worth a try.
Indeed, such as Kaiser's glycerinegelatine (7g gelatine, 42ml water, 50ml glycerin + a drop of conservant which used te be phenol but is now something else [what?]). Fern spores, much the same as pollen but not exactly, tend to swell in pure glycerine, sometimes a lot, and often burst shedding their contents. Here for temporary slides I prefer just soft gelatine.
--- felix filicis ---

Walter Piorkowski
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Re: A Pollen Project. Part 4. The Carnation

Post by Walter Piorkowski »

Thank you both for the good tips. I'll put them to good use in the future.
Walt

rjlittlefield
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Re: A Pollen Project. Part 4. The Carnation

Post by rjlittlefield »

iconoclastica wrote:
Fri Jan 24, 2025 2:42 pm
Kaiser's glycerinegelatine (7g gelatine, 42ml water, 50ml glycerin + a drop of conservant which used to be phenol but is now something else [what?]).
In place of the phenol...

https://www.microbehunter.com/microscop ... .php?t=686 suggests Listerine (a commercial mouthwash).

Commercial formulations of Kaiser's list a variety of preservatives: quaternary ammonium compounds, thimerosal, "proprietary", etc.

ChatGPT (4o) suggests thymol, sodium benzoate, methylparaben, and propylparaben (listing only common materials added in tiny amounts).

--Rik

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