Pediastrum

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hkv
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Pediastrum

#1 Post by hkv » Sun Jan 15, 2017 9:47 pm

Pediastrum, might be Pediastrum Integrum.

1st image. 100X DIC Oil. Stack of 4.

ImagePediastrum Integra by Håkan Kvarnström, on Flickr

2nd image. 40X DIC + 1.6X (Mag changer). Stack of 11.

ImagePediastrum Integra by Håkan Kvarnström, on Flickr

3rd image. 40X DIC + Autofluorescence in combination. Single image

ImagePediastrum Integra by Håkan Kvarnström, on Flickr

4th image. 100X Autofluorescence. Stack of 2.

ImagePediastrum Integra by Håkan Kvarnström, on Flickr

5th image. 100X DIC. Stack of 4.

ImagePediastrum Integra by Håkan Kvarnström, on Flickr
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75RR
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Re: Pediastrum

#2 Post by 75RR » Sun Jan 15, 2017 10:20 pm

Great images!
Do like Pediastrum, this reminded me that I have not seen one in quite a long time.
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Re: Pediastrum

#3 Post by kit1980 » Mon Jan 16, 2017 12:38 am

Beautiful! The second image is especially eye-pleasing.
What is this 1.6X (Mag changer)? Is this a feature of the stand?
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Re: Pediastrum

#4 Post by hkv » Mon Jan 16, 2017 7:35 am

Thanks!
kit1980 wrote:Beautiful! The second image is especially eye-pleasing.
What is this 1.6X (Mag changer)? Is this a feature of the stand?
Yes, it is a magnification changer mounted on the stand that give 1.25-2X extra magnification when needed.
Normally it is set to 1X and is then just a hollow tube.

It is the "round" box in the middle

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JimT
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Re: Pediastrum

#5 Post by JimT » Mon Jan 16, 2017 3:15 pm

They are all great. If I had to pick a favorite it would be the last one.
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Re: Pediastrum

#6 Post by rnabholz » Mon Jan 16, 2017 3:23 pm

Beautiful series.

Well done

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Re: Pediastrum

#7 Post by hkv » Mon Jan 16, 2017 10:03 pm

Thanks Jim and Rod! The last one is also my favourite.
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Johann
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Re: Pediastrum

#8 Post by Johann » Tue Jan 17, 2017 6:02 am

Nice find - great images :)
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gekko
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Re: Pediastrum

#9 Post by gekko » Tue Jan 17, 2017 5:00 pm

Excellent images! What excitation wavelength did you use? I assume that you used epi-illumnation for the excitation? If so, how would DIC work then? Please forgive me if my questions are too dumb... Also if you've already answered them elsewhere, in which case please only give the link (I have not been following the forum for quite some time, so I probably missed it).
Thanks for a very nice set of images.
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Re: Pediastrum

#10 Post by hkv » Tue Jan 17, 2017 7:57 pm

gekko wrote:Excellent images! What excitation wavelength did you use? I assume that you used epi-illumnation for the excitation? If so, how would DIC work then? Please forgive me if my questions are too dumb... Also if you've already answered them elsewhere, in which case please only give the link (I have not been following the forum for quite some time, so I probably missed it).
Thanks for a very nice set of images.
Excitation: Band Pass 400-440 nm.
Diachromatic mirror: 455 nm
Emission filter: Long Pass 475 nm

It is a standard Olympus U-MWBV2.

http://www.tengrant.com/upLoad/file/201 ... 715070.pdf

Yes, it is EPI-fluorescence. Exfo X-Cite with liquid light guide to the microscope EPI-illuminator.

The technique I used here (3rd image) is to use weak fluorescence light (EPI) in combination with weak ordinary halogen lightning from below through the condenser. So in practice the subject is illuminated from below and above simultaneously. The gives a weak DIC image where the autofluorescence color shines through in the final image. It is no problem to do EPI fluorescence with all the DIC prisms and polarizers installed. They seem to be transparent for a wide range of spectra.
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Re: Pediastrum

#11 Post by gekko » Thu Jan 19, 2017 11:22 pm

Many thanks for taking the time to explain in detail. Not much leeway between excitation and emission ranges. Nice sharp cut-off filters.
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