A very quick & easy water-mounted pollen stack

Here you can post pictures and videos to show others.
Post Reply
Message
Author
User avatar
mrsonchus
Posts: 3813
Joined: Tue Feb 03, 2015 9:42 pm
Location: Cumbria, UK

A very quick & easy water-mounted pollen stack

#1 Post by mrsonchus » Fri Apr 22, 2016 11:04 pm

Hi all, here's a quick example I made while looking at some 'Colt'sfoot' capitula today - handy that there's a nice big patch growing on a grass verge near the supermarket.
My Darling Wife went into the supermarket - I 'looked after the dog' - dragged her over to the Colt'sfoot and filled a 'pooh-bag' with a few nice flower-heads and seed-bearing heads! I did have several plants (from our last visit) in a trough in the garden, but I was 'persuaded' to remove them and plant 'proper flowers' instead.... :oops:

Anyway, placed several of the (dozens of) tiny flowers that make up the Colt'sfoot's composite flower-head into some tap-water on a slide, coverslipped and squashed lightly to remove air and get the coverslip to 'grip the water' as it were.

This is a stack of about 6 images taken through the 'scope with my 2mp 'Toupcam' - stacked in the super 'Toupview' software that complements the T-cam so well. The whole process from bag to image took about 10 minutes. Simply-done is I think the way to begin...
ws-x60-tussilago_farf.jpg
ws-x60-tussilago_farf.jpg (125.34 KiB) Viewed 2062 times
An easy water-mount, very few images (5 or 6 as I remember) in the stack and the use of the superbly efficient, totally free and easy-to-use 'Toupview' (you can load any images into it, no camera needs to be connected if preferred).

Measurements,
ws_coltsfoot-pollen-x40.jpg
ws_coltsfoot-pollen-x40.jpg (89.34 KiB) Viewed 2047 times
Last edited by mrsonchus on Sat Apr 23, 2016 4:51 am, edited 1 time in total.
John B

User avatar
rnabholz
Posts: 3079
Joined: Thu Jan 08, 2015 10:11 pm
Location: Iowa USA
Contact:

Re: A very quick & easy water-mounted pollen stack

#2 Post by rnabholz » Sat Apr 23, 2016 3:06 am

Very good image John.

I have recently had pollen suggested to me as a "must try" subject. Your image certainly reinforces that!

Thanks for the inspiration.

User avatar
mrsonchus
Posts: 3813
Joined: Tue Feb 03, 2015 9:42 pm
Location: Cumbria, UK

Re: A very quick & easy water-mounted pollen stack

#3 Post by mrsonchus » Sat Apr 23, 2016 3:49 am

rnabholz wrote:Very good image John.

I have recently had pollen suggested to me as a "must try" subject. Your image certainly reinforces that!

Thanks for the inspiration.
Go-get it Rod! :D

A little tip, 'dunk' the anther with it's pollen into your chosen medium, water, 50/50 water glycerin, 25%OH etc.. Air bubbles are far easier to eliminate or render harmless (i.e. out of the field of view) that way. Pollen on it's own can misbehave! :D

Good hunting. :)

Oh yes, the yellow 'oily glue' that often covers pollen and can cause problems optically is called 'pollenkit' and may be removed sufficiently with an OH wash if preferred. I didn't need to here as it separated of it's own accord with this pollen. Here's a picture of some of it on the coverslip's underside, out of the field of view. It and air bubbles etc moved away as I (gently) pressed the coverslip with the rubber bulb of a small pipette (they're ideal as they don't slip!)...

Pollenkit.....
ws_coltsfoot-pollenkit.jpg
ws_coltsfoot-pollenkit.jpg (132.09 KiB) Viewed 2049 times
John B

User avatar
gekko
Posts: 4701
Joined: Sun Oct 12, 2014 7:38 am
Location: Durham, NC, USA.

Re: A very quick & easy water-mounted pollen stack

#4 Post by gekko » Sat Apr 23, 2016 8:35 am

Very nice.

User avatar
rnabholz
Posts: 3079
Joined: Thu Jan 08, 2015 10:11 pm
Location: Iowa USA
Contact:

Re: A very quick & easy water-mounted pollen stack

#5 Post by rnabholz » Sat Apr 23, 2016 4:44 pm

Thanks for the tip John.

I'll see what I can do.

Rod

glennbech
Posts: 55
Joined: Thu Apr 07, 2016 10:34 pm

Re: A very quick & easy water-mounted pollen stack

#6 Post by glennbech » Sat Apr 23, 2016 5:01 pm

That's very nice! As I mentioned, I am new to this, and just wonder if there is a good reason to water mount the pollen. Would a "dry" mount work just as well? Just the pollen and a cover slip?

User avatar
mrsonchus
Posts: 3813
Joined: Tue Feb 03, 2015 9:42 pm
Location: Cumbria, UK

Re: A very quick & easy water-mounted pollen stack

#7 Post by mrsonchus » Sat Apr 23, 2016 8:11 pm

glennbech wrote:That's very nice! As I mentioned, I am new to this, and just wonder if there is a good reason to water mount the pollen. Would a "dry" mount work just as well? Just the pollen and a cover slip?
Unfortunately not Glen - dry pollen + coverslip would be very sub-optimal. The idea of my water-mount is to get quickly and easily as close as practically possible to a fully-mounted specimen with a high RI mountant and level coverslip touching the specimen. It's a compromise but does have some use and merit for a quick taxonomic image for example - a collection of such images alone is a wonderful resource.

Have a go at different approaches - resist going above x40 objective though, any higher and the working-distance will become perilously small without a good mount and damage could easily result.

Have you looked at the late great Walter Dioni's articles on this website - his 'onion epidermis series' is simply a superb guide to starting out - I started with these (and a lot of kind and knowledgeable advice from fellows in this forum) from scratch as a total newbie such as you are now Glen.

Good luck, keep us posted - start experimenting, you'll learn very quickly. :)
John B

User avatar
billben74
Posts: 1020
Joined: Sat Dec 26, 2015 10:33 pm
Location: Cambridge, UK

Re: A very quick & easy water-mounted pollen stack

#8 Post by billben74 » Sat Apr 23, 2016 11:29 pm

Really nice pollen pick.
Now that spring has sprung this seems like a nice relaxing subject.


...except now I've thought of trying to catalogue different plants' pollen...
I really must learn to think small one of these days...

Yet again mrsonchus serves up nice picks and inspiration too :)

User avatar
mrsonchus
Posts: 3813
Joined: Tue Feb 03, 2015 9:42 pm
Location: Cumbria, UK

Re: A very quick & easy water-mounted pollen stack

#9 Post by mrsonchus » Sun Apr 24, 2016 12:02 am

billben74 wrote:Really nice pollen pick.
Now that spring has sprung this seems like a nice relaxing subject.


...except now I've thought of trying to catalogue different plants' pollen...
I really must learn to think small one of these days...

Yet again mrsonchus serves up nice picks and inspiration too :)
Haha Bill, I've been looking at some rather unusual embryos today from obviously fertilized ovules taken from what My Darling Wife informed me were snowdrops... They've a most interesting 'hooked' embryo, the hook is even slightly evident in the dried seed at maturity it would seem. These I may well put through my 'extreme 10hr' protocol in the next week or so (they'll need 48hrs fixation first) - should make nice sections I think.
ws_snowdrop-embryo.jpg
ws_snowdrop-embryo.jpg (86.75 KiB) Viewed 2014 times
Any sections from 'Slicer' coming up old chap? I'd love to see how you're getting on. :)
John B

User avatar
mrsonchus
Posts: 3813
Joined: Tue Feb 03, 2015 9:42 pm
Location: Cumbria, UK

Re: A very quick & easy water-mounted pollen stack

#10 Post by mrsonchus » Sun Apr 24, 2016 12:13 am

gekko wrote:Very nice.
Thanks Gekko, pollens a charming subject I think - I really like the broken spike of one of the grains - reminds me somehow that they're a natural product rather than another man-made piece of plastic. :)

I love pollen, so quick and easy, with so much variety.. :D
John B

User avatar
KurtM
Posts: 1605
Joined: Tue Jun 09, 2015 12:08 am
Location: League City, Texas
Contact:

Re: A very quick & easy water-mounted pollen stack

#11 Post by KurtM » Sun Apr 24, 2016 12:42 am

mrsonchus wrote:A little tip, 'dunk' the anther with it's pollen into your chosen medium, water, 50/50 water glycerin, 25%OH etc..

Oh yes, the yellow 'oily glue' that often covers pollen and can cause problems optically is called 'pollenkit' and may be removed sufficiently with an OH wash if preferred.
I'm who suggested that Rod check out pollen, when he said he was thinking about going diatoming. I enthusiastically agree, and love your choice of words: pollen is charming study! I also love your tip for dipping anthers into medium, because, as you say, pollen grains tend to misbehave ... or stampede on you, as we say here in Texas.

But what's OH? And how do you wash pollen in it? I feel like I should know this, but I'm drawing a blank...
Cheers,
Kurt Maurer
League City, Texas
email: ngc704(at)aol(dot)com
https://www.flickr.com/photos/67904872@ ... 912223623/

User avatar
mrsonchus
Posts: 3813
Joined: Tue Feb 03, 2015 9:42 pm
Location: Cumbria, UK

Re: A very quick & easy water-mounted pollen stack

#12 Post by mrsonchus » Sun Apr 24, 2016 12:53 am

billben74 wrote:Really nice pollen pick.
Now that spring has sprung this seems like a nice relaxing subject.


...except now I've thought of trying to catalogue different plants' pollen...
I really must learn to think small one of these days...

Yet again mrsonchus serves up nice picks and inspiration too :)
Now then Bill, you've probably seen these before, but I've had a quick look through some of my old pollen adventures, how about starting that collection......

from some purple Crocus in our garden last year...
ws_crocus_pollen_detail.jpg
ws_crocus_pollen_detail.jpg (178.97 KiB) Viewed 2007 times
from a week's 'holiday' in Scotland on the coast last year - I spent a week studying a fine population of 'Red Campion' (Silene.dioica) growing in the cottage's garden - turned out they are dioecious and have male & female flowers on male and female plants!
ws_red_campion_pollen-0001.jpg
ws_red_campion_pollen-0001.jpg (44.66 KiB) Viewed 2007 times
this is Epilobium pollen ('Willowherb') with it's viscin-threads - sticky 'tendrils', these pollen grains stay in and are released as tetrads, unlike for example the above which are released as separate grains.. they're all so varied - fascinating things for sure! :D
ws_epilobium_pollen-(3).jpg
ws_epilobium_pollen-(3).jpg (163.33 KiB) Viewed 2007 times
here are some stained grains to finish off, first a Lily grain stacked to bring out the marvellous exine-ornamentation
ws_pollen_grain_2.jpg
ws_pollen_grain_2.jpg (85.63 KiB) Viewed 2007 times
and my study - ongoing - Sonchus ('Sowthistle') pollen 2-stained to contrast the body of the grain with its extraordinarily intricate exine.... my favourite pollen but very tough and brittle - difficult to section well,
ws_x100_2_stain_pollen_stac.jpg
ws_x100_2_stain_pollen_stac.jpg (374.34 KiB) Viewed 2007 times
There's a fascinating cornucopia of pollen waiting to be admired and collected! You know you want to....... just a few teeny grains on a slide to start with, maybe a little Fuchsine-treated glycerin-gel mountant...... :D :D
John B

User avatar
mrsonchus
Posts: 3813
Joined: Tue Feb 03, 2015 9:42 pm
Location: Cumbria, UK

Re: A very quick & easy water-mounted pollen stack

#13 Post by mrsonchus » Sun Apr 24, 2016 1:07 am

KurtM wrote:
mrsonchus wrote:A little tip, 'dunk' the anther with it's pollen into your chosen medium, water, 50/50 water glycerin, 25%OH etc..

Oh yes, the yellow 'oily glue' that often covers pollen and can cause problems optically is called 'pollenkit' and may be removed sufficiently with an OH wash if preferred.
I'm who suggested that Rod check out pollen, when he said he was thinking about going diatoming. I enthusiastically agree, and love your choice of words: pollen is charming study! I also love your tip for dipping anthers into medium, because, as you say, pollen grains tend to misbehave ... or stampede on you, as we say here in Texas.

But what's OH? And how do you wash pollen in it? I feel like I should know this, but I'm drawing a blank...
Hi Kurt, OH is just shorthand for alcohol - sorry about that. I wash grains using the anther as a 'holding-device' if you like, it's possible to do almost anything with the pollen 'stuck to something' such as this, although sticking grains to something else would also work, even something like a tiny piece of cloth would probably work - simply 'release' the grains when treatments are completed and it's time to observe or mount etc... On it's own in a free state pollen is a nightmare!

A similar technique with actual cloth/mesh is used for extremely small tissue in histology practice also - cells/tissue are 'wrapped' then processed right through all stages before being released for study.

If OH is ineffective try something like 'Wintergreen oil' (Methyl salicylate) as it's a good organic solvent also.. :)
John B

User avatar
KurtM
Posts: 1605
Joined: Tue Jun 09, 2015 12:08 am
Location: League City, Texas
Contact:

Re: A very quick & easy water-mounted pollen stack

#14 Post by KurtM » Sun Apr 24, 2016 3:57 am

mrsonchus wrote:...sticking grains to something else would also work, even something like a tiny piece of cloth would probably work - simply 'release' the grains when treatments are completed...
How does "simply releasing" them work? I have problems releasing things almost constantly... :P
Cheers,
Kurt Maurer
League City, Texas
email: ngc704(at)aol(dot)com
https://www.flickr.com/photos/67904872@ ... 912223623/

User avatar
mrsonchus
Posts: 3813
Joined: Tue Feb 03, 2015 9:42 pm
Location: Cumbria, UK

Re: A very quick & easy water-mounted pollen stack

#15 Post by mrsonchus » Sun Apr 24, 2016 4:38 am

KurtM wrote:
mrsonchus wrote:...sticking grains to something else would also work, even something like a tiny piece of cloth would probably work - simply 'release' the grains when treatments are completed...
How does "simply releasing" them work? I have problems releasing things almost constantly... :P

:D :D
John B

glennbech
Posts: 55
Joined: Thu Apr 07, 2016 10:34 pm

Re: A very quick & easy water-mounted pollen stack

#16 Post by glennbech » Sun Apr 24, 2016 10:55 pm

I think I saw something interesting when I observed the pollen of Alpine Pennygrass recently. The flower produces "fruit" that contains seeds in clusters. Almost like a raspberry I guess.

I was not aware of the fruit/seed production when I looked at my sample. But it sounds reasonable one plant sample could contain several fruit as well as pollen?

Well... off slide surfing again ... :)

/g

User avatar
mrsonchus
Posts: 3813
Joined: Tue Feb 03, 2015 9:42 pm
Location: Cumbria, UK

Re: A very quick & easy water-mounted pollen stack

#17 Post by mrsonchus » Sun Apr 24, 2016 11:12 pm

glennbech wrote:I think I saw something interesting when I observed the pollen of Alpine Pennygrass recently. The flower produces "fruit" that contains seeds in clusters. Almost like a raspberry I guess.

I was not aware of the fruit/seed production when I looked at my sample. But it sounds reasonable one plant sample could contain several fruit as well as pollen?

Well... off slide surfing again ... :)

/g
Certainly does Glen, it's probably one of many strategies that plants have to avoid self-pollination - in this case the timing of the production of the plant's pollen is after the receptive period during which the plant's stigmas will accept and admit pollen (from other plants as it's own pollen has not yet been released....
Sounds morphologically similar to the truly bizarre flowers of some Spurges, I've such a beast in my graden (a 'petty-spurge' or Euphorbia.peplus) - the fruits are 'nutlets' as I remember...

Other strategies include raising stigmas above the anthers, chemical prevention and consequent incompatibilty etc - some plants even have 'male only' and 'female only' flowers on the same or even separate individuals - fascinating! :D

I'll have a look when I get the chance, sounds interesting! :D
John B

User avatar
gekko
Posts: 4701
Joined: Sun Oct 12, 2014 7:38 am
Location: Durham, NC, USA.

Re: A very quick & easy water-mounted pollen stack

#18 Post by gekko » Mon Apr 25, 2016 6:48 pm

Great images and a lot to learn!

glennbech
Posts: 55
Joined: Thu Apr 07, 2016 10:34 pm

Re: A very quick & easy water-mounted pollen stack

#19 Post by glennbech » Wed Apr 27, 2016 9:12 am

mrsonchus wrote:
glennbech wrote:I think I saw something interesting when I observed the pollen of Alpine Pennygrass recently. The flower produces "fruit" that contains seeds in clusters. Almost like a raspberry I guess.

I was not aware of the fruit/seed production when I looked at my sample. But it sounds reasonable one plant sample could contain several fruit as well as pollen?

Well... off slide surfing again ... :)

/g
Certainly does Glen, it's probably one of many strategies that plants have to avoid self-pollination - in this case the timing of the production of the plant's pollen is after the receptive period during which the plant's stigmas will accept and admit pollen (from other plants as it's own pollen has not yet been released....
Sounds morphologically similar to the truly bizarre flowers of some Spurges, I've such a beast in my graden (a 'petty-spurge' or Euphorbia.peplus) - the fruits are 'nutlets' as I remember...

Other strategies include raising stigmas above the anthers, chemical prevention and consequent incompatibilty etc - some plants even have 'male only' and 'female only' flowers on the same or even separate individuals - fascinating! :D

I'll have a look when I get the chance, sounds interesting! :D
I got curious about this and took a look at my Alpine pennycress with my vintage Carl Zeiss Jena Stativ VA (from 1910ish) that has an objective that can do reflected light observations. It looks like some of the flowers on the plant expose the stigma but keep the stamen within the petals. I don't know if this is a part of the self-pollenation prevention. Maybe they flowers only open up when pollinated?

But. Now am derailing your thread: -)


Image

User avatar
mrsonchus
Posts: 3813
Joined: Tue Feb 03, 2015 9:42 pm
Location: Cumbria, UK

Re: A very quick & easy water-mounted pollen stack

#20 Post by mrsonchus » Wed Apr 27, 2016 11:31 pm

Hmm, it looks to me like that situation is really by virtue of the flower being partly open. When fully open it seems that the anthers and their pollen-load are definitely within reach of the stigma as they (the stigmas) aren't above them or distant from them in any way really. Perhaps this would signify that there's a chemical restriction in place to preclude self-pollination, this is very commonly the case and is a fascinating subject. Without the right chemical signals between pollen-grain and stigmatic surface germination and consequent penetration by the emerging pollen-tube is not possible. Amazing! :D

Beautiful anthers, such colour!

Here're a couple of superb papers re self-incompatibility mechanisms...
http://aob.oxfordjournals.org/content/8 ... 5.full.pdf
and
http://dev.biologists.org/content/devel ... 1.full.pdf

Fascinating! :)
John B

Post Reply