Dragging detail from pollen sections - filters & PP

[mrsonchus]

Hi all, I've been slide-surfing with the big-O and giving the x63 Fl 160mm 0.95 objective a run-out. Looking over some un-labelled slides I came across one of Sonchus.oleraceus ('Smooth Sow-Thistle' - a common weed in the UK) pollen, sectioned at 4µ and stained with Safranin (for the nuclei) and Fast-Green (for everything else....), permanently mounted in 'Omnimount' resinous mountant.

Anyway I had a go at using a green filter over the light-source before setting custom WB with the Canon 1200D used, and stacking 5 images to try to get as much information from the slide as I could. The idea being that the green filter would increase detail and contrast, while the PP may help further by converting to B & W then inverting - always good to bring out details I find. Also I rather rashly added an 'emboss' effect just to see what it looks like.

Here are a few pictures, quite a lot of detail has indeed been revealed re the structure of the grains, both of the external 'Exine' - the spiny case that covers these grains, and the internal nuclei of this almost-mature (micro) gametophyte - the first nucleus. the large 'tube nucleus' that builds the pollen-tube is there of course, also two sperm nuclei are seen - although only one discrete cell exists - it just has three nuclei!

Here are a few images you may find interesting,
This is the 'normal' as-on-slide image that has good exine detail,

ws_x63 gr filter 3stack.jpg (39.99 KiB) Viewed 2682 times

and after a rather dodgy application of emboss....

ws_PP textured x63 gr filter 3stack.jpg (39.54 KiB) Viewed 2682 times

Another 'normal' stained-slide image that shows the three nuclei, stained red by the Safranin stain used,

ws_x63 gr filter 5stack copy.jpg (35.52 KiB) Viewed 2682 times

Here after 'some treatment' - see what you think about detail enhancement, I can definitely see some,

ws_x63 gr filt 5stack inverted.jpg (55.43 KiB) Viewed 2682 times

Finally here's a section that is right through the middle of a grain (these grains are bout 30µ in diameter and are sectioned here at 4µ. This is an 'equatorial plane' which shows some very nice geometry of the grain, better than a whole grain focused to the equatorial plane in this respect....

ws_x63 single gr filter.jpg (45.79 KiB) Viewed 2682 times

I like this slide as it has about 200 such grains within and I'm going to put it back into my 'keeper' slide cases...

Great fun to dig-out the old 'waste slide' boxes and have a look at them - you never know what's in there. Hope you find these interesting - just a small exercise this evening to practice with the Orthoplan!

John B

Just found an old post that features this slide! Also a nice 15µ slide that really shows the 3D exine structure well,

here's the link --> Original post with this slide

[billbillt]

Yes Sir, John B.!!... You have done it again!... Another wonderful series of an interesting subject!..

BillT

[zzffnn]

Nice work, John B.

I wonder how "emboss" works in post processing. If it works as the name suggested, it may overemphasize thin stained areas with textural relief, while ignoring thick transparent areas?

[mrsonchus]

Nice work, John B.

I wonder how "emboss" works in post processing. If it works as the name suggested, it may overemphasize thin stained areas with textural relief, while ignoring thick transparent areas?

Hmm, I don't know zz' - an interesting question. I would be a pretty good exercise to look into these PP techniques in an organized way sometime, as they clearly have a place when the main criterion is the revelation of details for taxonomic comparisons for example...

Even the closely related Sonchus oleraceus and asper species show quite a lot of morphological difference in their pollen - not radically so but quite evident under close scrutiny. It's not easy however to get comparable views of the respective grains, hence the adoption I suppose of the 'std' views such as the polar-plane, sectioned or not.

It's a really interesting area, in fact the first I ever tried when I started with microscopy - everyone is easily able to get some pollen onto a slide for a good initial run-out of their 'scope!

This is also a good time of year for Botany the UK as everything is just starting to come out now - early Spring here...

Thanks for looking my friend, John B

[FungusMan]

What is your x63 objective please?

[mrsonchus]

What is your x63 objective please?

Hi, that objective (sold back in 2019 together with a superb Orthoplan when I upgraded to an Olympus BX40) was a surprisingly good performer. The Orthoplan is nominally a 170mm TL 'scope and the 63x is a 160mm TL objective for use with the smaller, later 160mm Leitz models I believe.
Anyway, as I've always wanted to be able to reach-in a little further than 40x whilst not taking the observation over to oil-immersion, with just about every 'scope capable I've looked for some kind of 60x-ish objective.
As I say this nice 63x Fl 0.85 worked rather well with the Orthoplan and filled said niche for me quite nicely.

Here are a couple of images of it from 2019,


and

[FungusMan]

Thanks for your explanation. Yes, I agree with you, it is a great performer. Images are pretty detailed with above-average resolution.

[DonSchaeffer]

Great microscopy!!