Spirogyra and Trachelomonas

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75RR
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Spirogyra and Trachelomonas

#1 Post by 75RR » Thu Dec 18, 2014 3:26 pm

16x objective, alga width 50µm, Trachelomonas 20µm, Darkfield.
Apart from what some might construe as a seasonal image – reddish, green and twinkly things – anyone know what the moving specs within the alga envelope are?

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vasselle
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Re: Spirogyra and Trachelomonas

#2 Post by vasselle » Thu Dec 18, 2014 4:08 pm

Bonjour 75RR.
Très belle vidéo.
Ce que l'on vois bouger ce sont des cristaux de baryum.
Cordialement seb
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Re: Spirogyra and Trachelomonas

#3 Post by gekko » Thu Dec 18, 2014 6:54 pm

Very nice video! I don't know if this helps answer your question: http://jcb.rupress.org/content/106/5/1545.abstract
I have taken a video of the brownian motion of those particles that shows up very nicely in cross-polarized light, and if I may be so bold, I would suggest that next time you get a nice alga under your objective, perhaps you may want to consider cross-polarized light (that ususally gives dramatic results in green algae) in addition BF, oblique or DF.

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75RR
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Re: Spirogyra and Trachelomonas

#4 Post by 75RR » Thu Dec 18, 2014 8:06 pm

Thanks vasselle and gekko.

gekko, that link went right over my head :)
Managed to dig out the phrase "cytoplasmic streaming". I take it that could account for the movement?

Perhaps I should have tried additional techniques, problem is so far I only have BF, DF and Offset DF (Oblique).

Hope to add Phase soon, picked up an old Phase condenser the other day that will go on my Zeiss Standard RA which is almost ready, including "inventive" trinocular head, just awaiting light.
... perhaps you may want to consider cross-polarized light ...
Is that not a major expensive upgrade/dedicated microscope?
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Re: Spirogyra and Trachelomonas

#5 Post by gekko » Thu Dec 18, 2014 8:37 pm

75RR wrote:Thanks vasselle and gekko.

gekko, that link went right over my head :)
Managed to dig out the phrase "cytoplasmic streaming". I take it that could account for the movement?

Perhaps I should have tried additional techniques, problem is so far I only have BF, DF and Offset DF (Oblique).

Hope to add Phase soon, picked up an old Phase condenser the other day that will go on my Zeiss Standard RA which is almost ready, including "inventive" trinocular head, just awaiting light.
... perhaps you may want to consider cross-polarized light ...
Is that not a major expensive upgrade/dedicated microscope?
Good luck on both light and phase! No, polarized light is very inexpensive. All it involves is a couple of linear polarizing filters (old camera polarizers, even plastic polarizing sunglass will do) or filters cut from inexpensive polarizing plastic sheet. If you have a camera linear polarizing filter you can put it over the light port where you can rotate it to get extinction. The other one (analyzer) goes in any convenient place between objective and eye (or camera) but I would avoid a conjugate of the image plane in order to avoid dust & scratches on the filter being visible. Most convenient place might be between the objective turret and the binocular/trinocular head (and it can be left there even when not using cross-polarized light-- its side effect will be to reduce the light level only slightly). This is best if it is very thin, as adding a thick glass or plastic filter there will slightly alter the effective tube length and may introduce some spherical aberration (one advantage of infinity optics but only if you can place it between objective and tube lens). I understand that one can remove the polarizing thin plastic film from a (damaged) LCD display (I never tried it so I may be wrong), and that may be nice and thin to use as analyzer. For a retarder over the bottom polarizer (to get "nice" colors) you can try any transparent plastic piece (e.g., broken CD "jewel" case, cellophane wrapper, etc.) and rotate it for different effects. Seb (Vasselle) uses a circular polarizer for the bottom filter as both polarizer and retarder, but I've not had success with one (I tried both directions).

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Re: Spirogyra and Trachelomonas

#6 Post by vasselle » Fri Dec 19, 2014 2:03 pm

Bonjour 75RR
Pour me filtre polarisant j'utilise un filtre polarisant d'un objectif photo par dessus la lumière de microscope puis je mes par dessus du filtre polarisant un morceau de plastique genre CD et après je remet un autre filtre polarisant.
Et les filtres qui fonctionne bien pour faire un filtre polarisant ce sont les écrans des téléphones portable mes par contre il faut retiré la fine couche qui il y a dessus et la garder car c'est cette fine couche que l'on fait le filtre polarisant et ça fonctionne très bien.
Dans la pratique le mieux c'est essayer différentes technique et voir ce que ça donne.
Bonne observation
Cordialement seb
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Re: Spirogyra and Trachelomonas

#7 Post by 75RR » Fri Dec 19, 2014 4:52 pm

Thanks gekko and vasselle, will look into it.
I suspect the top polarizing filter (the one above the objective) will be the challenge.
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Re: Spirogyra and Trachelomonas

#8 Post by vasselle » Fri Dec 19, 2014 5:30 pm

Bonjour 75RR
Demain je te posterais quelques photos du montage que j'utilise sur mon microscope.
Comme ça si ça peux aider.
Cordialement seb
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75RR
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Re: Spirogyra and Trachelomonas

#9 Post by 75RR » Fri Dec 19, 2014 5:33 pm

Demain je te posterais quelques photos du montage que j'utilise sur mon microscope.
Thanks. That would be helpful.
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Re: Spirogyra and Trachelomonas

#10 Post by vasselle » Mon Dec 22, 2014 6:25 pm

Bonjour 75RR
Voici en photo le filtre polarisant que j'utilise pour obtenir des fonds colorer etc..
Pour le filtre polarisant je le mets sur la base de éclairage du microscope.
Et par dessus du filtre je mets un morceau de plastique genre pochette de CD.
Il y a aussi les écrans des téléphones portable qui fonctionne très bien ailleurs sur la première photo on en vois deux de filtre polarisant fait avec des écrans de téléphones portables.

Photo 1.
IMG_0338 (Copier).JPG
IMG_0338 (Copier).JPG (96.82 KiB) Viewed 6436 times
Photo 2.
IMG_0341 (Copier).JPG
IMG_0341 (Copier).JPG (123.79 KiB) Viewed 6436 times
Photo 3.
IMG_0340 (Copier).JPG
IMG_0340 (Copier).JPG (119.11 KiB) Viewed 6436 times
Photo 4.
IMG_0342 (Copier).JPG
IMG_0342 (Copier).JPG (100.13 KiB) Viewed 6436 times
J'espère que cela pourra aider .
Bonne observation
Cordialement seb
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Boitier EOS 1200D + EOS 1100D

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75RR
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Re: Spirogyra and Trachelomonas

#11 Post by 75RR » Mon Dec 22, 2014 10:07 pm

Hi vasselle,
Thank you for posting the photos.
I had understood that the second polarizing filter had to be somewhere in between the objective and the eye/camera.
I take it that you have found that placing both (one above the other) between the field diaphragm and the objective also works well.
On the lookout for a CPL filter at the moment.
Thanks again.
Zeiss Standard WL (somewhat fashion challenged) & Wild M8
Olympus E-P2 (Micro Four Thirds Camera)

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Re: Spirogyra and Trachelomonas

#12 Post by JimT » Mon Dec 22, 2014 11:38 pm

Here is another possibility from this site:
http://www.microbehunter.com/simple-pol ... icroscopy/
I really don't want to take apart one of my camera polarizers or break my sun glasses so I haven't done it yet.
I am going to look for polarizing plastic sheet.

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Re: Spirogyra and Trachelomonas

#13 Post by 75RR » Mon Dec 22, 2014 11:59 pm

Thanks for the link JimT

Interestingly the second filter is also below the objective but in this case above the specimen.
I am going to look for polarizing plastic sheet.
I think you are right, cheap and cheerful is the way to go.
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Re: Spirogyra and Trachelomonas

#14 Post by gekko » Tue Dec 23, 2014 12:33 am

My 2-cents' worth: In theory, the bottom polarizer ("polarizer") can be anywhere between the light source and the specimen; the top polarizer ("analyzer") can be anywhere between the specimen and the eye or camera sensor. But I think it is quite impractical to place the analyzer between the slide and the objective except when a low power or long working distance objective is used, and it is not a good place anyway because it is close to the image plane (so dust and scratches on the filter may mar the image). More importantly, if the objective specifies a cover glass thickness, this would be upset by the added thickness of the filter. I think by far the best and most convenient place for the analyzer is under the microscope "head" (if the monocular/bninocular/trinocular) head is removable. And it can stay there semi-permanantly for convenience. Another would be on top of the eyepiece or projection lens. The analyzer could be placed between the condenser and the slide, but that again is rather awkward, and placing it under the condenser (over the light port, for example, where it can be easily rotated to get extinction) would be where I'd put it. Putting the two polarizing filters there on top of one another may be a good way to vary the light intensity (without changing the color temperature of the illumnation if one is using tungsten/halogen illumination) bit it will not work to get polarized light images (once crossed there will be virtually no light going to the specimen, anyway).

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