A Mixture of Things Within Moss

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mrsonchus
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A Mixture of Things Within Moss

#1 Post by mrsonchus » Sat Sep 28, 2019 3:45 pm

Hi all, while I was examining some mosses this morning I thought I'd 'hook-out' one of the ubiquitous cyanobacteria 'globules' so often found in moss.
Anyway, this seemed a good time to have a practice with the darkfield and phase capabilities of the condenser of my Olympus BX40. I don't have much use for these techniques but why not have a go at them if they're there....
The Objective for the Ph2 images is a simple Olympus Achr 40x Ph2 objective. I actually quite like it's performance although I have very little phase experience. The phase contrast of my old Orthoplan never really seemed to 'cut the mustard' with me for some reason, aesthetic probably rather than technical.... As soon as I tried the phase of the BX40, with this simple Achr objective, I preferred the look of the phase images, and do now occasionally use phase for live temporary-slide examination.

Here are a few of the DF images using the darkfield position on the phase condenser, of the somewhat flattened and coverslipped cyanobacteria 'blob' as above. Not very good images compared the the amazing DIC versions I know, but quite interesting to me as a brightfield practitioner. Water-mounted with coverslip.

Like a great-big bundle of beaded necklaces, the cyanobacteria from their gelatinous 'blob' found in my moss samples....
Image
and
Image

Other 'things' lurk also it seems, althoguh I don't actually know what they are apart perhaps from 'algae'?
Again in the simple darkfield of the Olympus phase condenser's DF position which I believe is usable to about 0.65 n.a..
Image

Here's a phase contrast 40x Ach objective version of the last image, maybe even a diatom or desmid-type thing in there?
Image

Another mixture, phase again,
Image

A 2-image stack, does anyone have an idea what the tiny black rods are in the image below?
Image

More beads, some larger ones in the chains?
Image

Some brightfield, a couple of objects with 'tails'....
Image
and
Image

Not very good images, but an idea, especially for a beginner, of just what may be found in a miniscule gelatinous 'blob' stuck in between moss leaves!
John B

mnmyco
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Re: A Mixture of Things Within Moss

#2 Post by mnmyco » Sat Sep 28, 2019 4:22 pm

The “beads” are probably cyanobacteria, such as Nostoc species.

Hobbyst46
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Re: A Mixture of Things Within Moss

#3 Post by Hobbyst46 » Sat Sep 28, 2019 4:53 pm

John B,
1. Especially the darkfield and phase contrast are just beautiful. Did you post process, any besides resize ?
2. The only thing that is missing here is the images is the moss... :)
3. I would risk a guess that the tiny black rods are bacteria.
Zeiss Standard GFL+Canon EOS-M10, Olympus VMZ stereo

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mrsonchus
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Re: A Mixture of Things Within Moss

#4 Post by mrsonchus » Sat Sep 28, 2019 6:31 pm

Hobbyst46 wrote:
Sat Sep 28, 2019 4:53 pm
John B,
1. Especially the darkfield and phase contrast are just beautiful. Did you post process, any besides resize ?
2. The only thing that is missing here is the images is the moss... :)
3. I would risk a guess that the tiny black rods are bacteria.
Hi Hobby' - yes I used a touch of 'auto-level' correction. I very often take images with lowered contrast as a camera-setting (I don't bother with RAW as for my type of subject the focus isn't critically sharp) to optimise detail retention. Then, the PP level auto-adjustment with restraint. No sharpening applied though as with this type of image I think it would be counter-productive.

For example the backgrounds of the darkfield images are easily darkened if necessary with an adjustment at the 'dark-end' of levels - namely ensuring that the darkest level in the image is at or very close to black, with a simple drag of the dark-end level slider in PSEv9 that I have used for years - no subscription post-purchase like today's PS versions.

Here's an out-of-camera phase image,
Image

This is after the aforementioned level adjustment to increase contrast, quite a difference, and I personally think very good for forum content viewed across so many different device screen-types and colour-spaces.
Image

This type of image/microscopy isn't my forte (if I have one!) as I use brightfield with prepared permanently-mounted & stained Botanical slides almost exclusively, but I obviously still look at live tissue frequently when preparing or selecting tissue for processing/microtomy.

That-said, I am beginning to consider the addition of DIC capability to my system - in fact I'm so impressed with the Olympus BX40 and it's UIS2 optics I'm mulling-over an upgrade to a more versatile stand such as the BX50 series, if I can spot perhaps a bare stand at a good price.
The DIC images that I've seen across this forum over the years have so-impressed me that I may even go for DIC in the not-too-distant future, after I've added polarisation, simple to start with then perhaps with various retardation-plates and/or analysers.

This thought beyond basic polarisation (i.e. fixed analyser and rotatable polariser over light-port) came when I read a little about the Olympus 'gout-kit', which enables the actual types of birefringent materials to to some degree be distinguished, as for example in the case of gout (which I have no interest in, just the principles).

Here's an excellent Nikon page re polarisation (and gout as it happens).

I discovered not long ago in fact the usefulness of polarisation, which I used to rather ignorantly think of as 'all about crystals', for examination of plant anatomy. The orientation of certain fibers, microfibrils etc within cell-walls is very effectively imaged with polarisation, and indeed phase-contrast! It seems that when the basic polarisation is built-upon, that a whole World of additional imaging opens-up.

Thanks for the feedback Hobby'! :D :D
John B

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75RR
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Re: A Mixture of Things Within Moss

#5 Post by 75RR » Sat Sep 28, 2019 6:46 pm

Great phase!
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Re: A Mixture of Things Within Moss

#6 Post by mrsonchus » Sat Sep 28, 2019 6:56 pm

mnmyco wrote:
Sat Sep 28, 2019 4:22 pm
The “beads” are probably cyanobacteria, such as Nostoc species.
Hi 'myco' - yes I found some information that details the association between Nostoc and mosses, in particular 'Feather-moss'.
My images appear to show (according to a quick search at least...) a motile stage called "hormogonia" - the shorter filaments, the free-living stage - the long filaments, the "heterocysts" - the larger 'beads' included in the filaments.....

Fascinating, and all from a 'blob' on a moss-leaf!
John B

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Re: A Mixture of Things Within Moss

#7 Post by mrsonchus » Sat Sep 28, 2019 7:03 pm

Here, especially for Hobby'!

A couple of images of this morning's moss, whence came the cyanobacteria colony 'blob' above....

View 1 of a lump of damp moss from a plantpot,
Image

View 2,
Image

View 3,
Image

Lovely! My advice to any beginner, go and get a lump like this, and explore.............. :D :D
(and of course let us know what you find!)
Last edited by mrsonchus on Sat Sep 28, 2019 7:07 pm, edited 1 time in total.
John B

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mrsonchus
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Re: A Mixture of Things Within Moss

#8 Post by mrsonchus » Sat Sep 28, 2019 7:05 pm

75RR wrote:
Sat Sep 28, 2019 6:46 pm
Great phase!
Thanks 75' - I'm really starting to like phase. :)
John B

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Re: A Mixture of Things Within Moss

#9 Post by Wes » Sat Sep 28, 2019 9:40 pm

Very nice images, I especially like the darkfield ones. Now I kind of want to get some moss too :)

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Re: A Mixture of Things Within Moss

#10 Post by mnmyco » Sun Sep 29, 2019 3:26 am

Keep an eye out for tardigrades in your moss.

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