Licmophora (one more)

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75RR
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Licmophora (one more)

#1 Post by 75RR » Wed Aug 14, 2019 10:56 am

Plan 40x/0.65, DIC, 300µm, stacked and stitched in Photoshop, Marine sample, Alboran Sea.

Slightly angled which gives a better idea of the structure
Attachments
163-254(2).jpg
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Last edited by 75RR on Thu Aug 15, 2019 1:54 pm, edited 1 time in total.
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Re: Licmophora (one more)

#2 Post by Hobbyst46 » Wed Aug 14, 2019 3:09 pm

A very nice image, again! from an angle that gives a 3D view.

I copied the image into my Irfanview software and zoomed in. The dots are fairly visible in what I believe are stria. Done some measurements and calculation, and again I find that there are 16stria/10um in the "head" and 17/10um in the tail. Not within the stated 23-27/10um for L. colosalis.
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Re: Licmophora (one more)

#3 Post by tgss » Wed Aug 14, 2019 4:39 pm

Another beautiful image - thanks again for posting!
Tom

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Re: Licmophora (one more)

#4 Post by 75RR » Wed Aug 14, 2019 8:10 pm

Many thanks Hobbyst46 and tgss
I copied the image into my Irfanview software and zoomed in. The dots are fairly visible in what I believe are stria. Done some measurements and calculation, and again I find that there are 16stria/10um in the "head" and 17/10um in the tail. Not within the stated 23-27/10um for L. colosalis.
Thanks. I haven't sent the other images in yet.

Looks like I am going to have to find another suspect. Can't think which, as at 300µm, I don't suppose there are that many candidates.
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Re: Licmophora (one more)

#5 Post by 75RR » Thu Aug 15, 2019 7:19 am

Here is another one at 365µm in Black and White

I have attempted to measure the stria in 10µm and get 27ish which is as defined for Licmophora colosalis - perhaps you can remeasure

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Re: Licmophora (one more)

#6 Post by Hobbyst46 » Thu Aug 15, 2019 9:43 am

Hi 75RR - this is becoming better every day!

1) The UV image is a revelation in itself. Irrespective of the following stria discussion, please, disclose how you iamged it at 365nm ? a new specific modification of the Zeiss setup, illumination etc ? is it still DIC ?

2) The UV image is surprising - how comes all stria are visible "at the expense" of the chloroplasts and other morphological features of the live diatom ? or, did you clean the diatom before imaging ?

3) Of course, I rushed to re-measure at all three-four photos.
The UV image gives the same figure as yours, 27stria/10um.
The valve view {file Licmophora(2)} still gives 15-16 stria/10um.
The girdle view {file Licmophora(1)}, gives a really very rough estimate of 12stria/10um.

So, perhaps the very beautiful presentations of the live diatom in plain visible light DIC provide an unreliable display of the stria. Sounds blasphemy/stupidity, I know. I doubt that this is a result of the different wavelength alone. UV should improve resolution relative to 550nm, but not that much .

Last, but most important - it is L Colosalis after all!
Last edited by Hobbyst46 on Thu Aug 15, 2019 9:53 am, edited 1 time in total.
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Re: Licmophora (one more)

#7 Post by Hobbyst46 » Thu Aug 15, 2019 9:51 am

75RR,

I have a crazy idea how to clean a single diatom and watching it all the way, from live diatom to a frustule or separate valves: build a microfluidic setup where the internal outflow tubings are of a smaller diameter than 20um (say). Catch a diatom, put it inside, and flush it with water, then chemicals, then water again, all under continuous microscopical inspection.
Piece of cake... :twisted: although doable.
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Re: Licmophora (one more)

#8 Post by 75RR » Thu Aug 15, 2019 9:56 am

Hi Hobbyst46,

this is the original source for the image - using 63x/1.4, DIC, 5 stacks stitched in photoshop (reduced in size and converted to jpeg - original is tiff and 316 MB)

I rotated image and converted to B/W to simplify.

Note: I might have got away with 4 stacks - but I tend to err on the side of caution. It is frustrating when images don't stitch!

Did not actively clean the sample - but it may have occurred naturally (i.e. died + bacterial action) as I have had this same sample jar on my desk for many days now.
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75-232(a) copy.jpg
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Re: Licmophora (one more)

#9 Post by Hobbyst46 » Thu Aug 15, 2019 10:22 am

In the "staircase" stitch photo, at the wider pole (on the right) I find 20+ stria/10um.
Seemingly, as Maria Dolores Belando and coworkers stated, it is difficult to identify... but I wonder, if this is an example that the apparent resolution of the stria (or even dots) in a live diatom can differ from that of the clean silica frustule/valve ?
Last edited by Hobbyst46 on Thu Aug 15, 2019 10:45 am, edited 1 time in total.
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Re: Licmophora (one more)

#10 Post by 75RR » Thu Aug 15, 2019 10:45 am

Note the the number of stria varies between areas with red circles and areas with a blue circle
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Re: Licmophora (one more)

#11 Post by Hobbyst46 » Thu Aug 15, 2019 10:53 am

Agreed, the variation is between 16 per circle and 18-19 per circle. In the literature I noticed that in some diatoms the stria density at the two poles (of Licmophoras) is the same whereas in others it is different, but by a factor of up to 1.5 or so.

By the way (I avoid "BTW" now!), how is the monochrome image related to 365um ?
Last edited by Hobbyst46 on Thu Aug 15, 2019 11:36 am, edited 1 time in total.
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Re: Licmophora (one more)

#12 Post by 75RR » Thu Aug 15, 2019 11:31 am

... how is the monochrome image related to 365nm ?
Not 365nm but 365µm
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Re: Licmophora (one more)

#13 Post by Hobbyst46 » Thu Aug 15, 2019 11:37 am

75RR wrote:
... how is the monochrome image related to 365nm ?
Not 365nm but 365µm
sorry about that. 365um, editted the above response.
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Re: Licmophora (one more)

#14 Post by 75RR » Thu Aug 15, 2019 12:55 pm

Added a blue and 2 yellow squares aligned with the stria and of more precise length than the previous one with the circles which was just to draw attention to the different type of stria in the central and the side bands.

Image
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Re: Licmophora (one more)

#15 Post by Hobbyst46 » Thu Aug 15, 2019 1:22 pm

75RR wrote:Added a blue and 2 yellow squares aligned with the stria and of more precise length than the previous one with the circles which was just to draw attention to the different type of stria in the central and the side bands.
Aha, I think that quite clarifies (pun) the situation. The density in the central part of the girdle view (blue square) is much lower than in the external parts of the "sandwich" (red squares). I also think (cannot prove) that the same area of low density of striae is visible in image 163-254(1), whereas in that photo on the whole, only a few striae are really visible.
So, consistently, the high striae density areas are invisible in the valve view of the live diatom.

So,
(1) we may safely assume that your initial identification of L. colosalis is appropriate.
(2) the images of the live and kicking 8-) diatom do not represent the true striae density in this case
(3) keeping the diatom in the wet mount trapped and pressed by the coverslip for a couple of days, until chloroplasts and mucilage deteriorate can remedy issue (2) above.

Thanks for initiating the mental adventure...
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Re: Licmophora (one more)

#16 Post by 75RR » Thu Aug 15, 2019 1:30 pm

So,
(1) we may safely assume that your initial identification of L. colosalis is appropriate.
:) :)
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