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Amoeba - Scanning Electron Microscope images

 
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Ecki



Joined: 13 Aug 2008
Posts: 773
Location: Cape Town, South Africa

PostPosted: Wed Oct 09, 2013 12:52 pm    Post subject: Amoeba - Scanning Electron Microscope images Reply with quote

<disclaimer mode on>
Looking at Scanning Electron Microscope images gives a very different perspective as the amount of detail is sometimes overwhelming. Also, the image is comparable to a reflected light image - no details of the inside are revealed.

As I have very little comparable reference images, it is hard to say whether things are artefacts of the preparation or "real".
<disclaimer mode off>

Having said all this, here are some images of an amoeba. The amoeba has a very interesting pattern on it's cell coating. Amoebas very often have stowaways, here a fairly large diatom. The smaller diatom in the foreground is just being eaten.






Looking at microbes from the top is pretty boring, you want to tilt the stage. This gives very nice and fascinating perspectives in very small dimensions: http://www.photomacrography.net/forum/viewtopic.php?t=20341&highlight=choano%2A

With the Scanning Electron Microscope, the beam comes from the top and the detector is at the side, at an angle of approx. 45°. To compare this with a light microscope, this is as if your light comes from 45° and your lens is parallel to the specimen. If you tilt the image in the direction of the detector (the light) there are areas that will have shadow. In addition to this, edges emit more electrons and thus tend to be overly bright which easily results in clipping. To make matters worse, when I sit in front of black and white images in a darkened room all evening, my sense for correct image brightness seems to get confused, too.

I am sometimes struggling with post processing of SEM images. I can take 256 bit TIFF images, up to approximately 3000x2000 pixels. Ideas and tips for post processing are highly appreciated. At the moment I am using Topaz details.

Anyhow, I hope you enjoy these images.

Best regards
Ecki
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Fredlab



Joined: 09 Nov 2012
Posts: 304
Location: Burgundy

PostPosted: Wed Oct 09, 2013 12:56 pm    Post subject: Reply with quote

wow...
amazing.

Thanks.
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ChrisR
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Joined: 14 Mar 2009
Posts: 7256
Location: Near London, UK

PostPosted: Wed Oct 09, 2013 1:09 pm    Post subject: Reply with quote

FredLab wrote:
wow...
amazing.

Thanks.

The English is perfect!!


Ecki - are you telling us you have a new toy in your shed?
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JH



Joined: 09 Mar 2013
Posts: 945
Location: Vallentuna, Stockholm, Sweden

PostPosted: Wed Oct 09, 2013 1:09 pm    Post subject: Reply with quote

Very interesting!

Regards Jörgen
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Bruce Taylor



Joined: 23 Jun 2011
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Location: Wakefield, Quebec / Ottawa, Ontario

PostPosted: Wed Oct 09, 2013 1:25 pm    Post subject: Reply with quote

Those images are just staggering, Ecki. Absolutely wonderful.
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leonardturner



Joined: 14 Mar 2013
Posts: 314
Location: Atlanta, GA, USA

PostPosted: Wed Oct 09, 2013 2:23 pm    Post subject: Reply with quote

Very good images. I can be of no help to you, as my SEM experience was many years back, but I am curious. How do you keep the amoeba from totally desiccating in the vacuum of the SEM? Does the gold coating persist even if the organism disappears?

Thanks--
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pwnell



Joined: 18 Dec 2009
Posts: 1939
Location: Tsawwassen, Canada

PostPosted: Wed Oct 09, 2013 5:37 pm    Post subject: Reply with quote

Really well done. I wish we have more SEM / TEM images - the details and 3D structures they reveal is just incredible. Keep them coming.
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RogelioMoreno



Joined: 20 Nov 2009
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Location: Panama

PostPosted: Wed Oct 09, 2013 6:12 pm    Post subject: Reply with quote

Ecki,

Very nice, a love the details, please more.

Rogelio
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Ecki



Joined: 13 Aug 2008
Posts: 773
Location: Cape Town, South Africa

PostPosted: Thu Oct 10, 2013 2:53 am    Post subject: Reply with quote

Quote:
Ecki - are you telling us you have a new toy in your shed?


The toy is not in my shed but in my new lab in the basement - on a 1800 kg block of concrete Mr. Green

Thanks for the nice comments and yes, there will be more.

Best regards
Ecki
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carlos.uruguay



Joined: 23 Feb 2012
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Location: Uruguay - Montevideo - America del Sur

PostPosted: Thu Oct 10, 2013 3:44 am    Post subject: Reply with quote

Impressive images, thank you for sharing!
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Jacek



Joined: 02 Oct 2011
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Location: Poland

PostPosted: Thu Oct 10, 2013 4:05 am    Post subject: Reply with quote

I really like high magnification is impressive
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Olympusman



Joined: 15 Jan 2012
Posts: 3422

PostPosted: Thu Oct 10, 2013 10:41 am    Post subject: Amoeba Reply with quote

Nice shots, but how do you preserve an organism that is over 90% water to be photographed in a vacuum? From what I know you infuse the protozoa with a uranium compund, but I find it hard enough to apply chemicals in light microscopy.
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g4lab



Joined: 23 May 2008
Posts: 1423

PostPosted: Fri Oct 11, 2013 11:38 am    Post subject: Reply with quote

Ecki's new toy is wonderful as are the images.

Is it an environmental SEM. Some of those have the capability to look at , for example , live specimens. They don't subject the specimen to high vacuum and don't require dessication.
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rjlittlefield
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Joined: 01 Aug 2006
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Location: Richland, Washington State, USA

PostPosted: Fri Oct 11, 2013 12:15 pm    Post subject: Reply with quote

Indeed a wonderful image.

I'm guessing freeze-dried.

Environmental SEMs are great for imaging the surface of subjects that have water in them, but they won't see through bulk water where the amoeba would be when alive.

--Rk
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Ecki



Joined: 13 Aug 2008
Posts: 773
Location: Cape Town, South Africa

PostPosted: Sat Oct 12, 2013 12:29 am    Post subject: Reply with quote

These are high vacuum mode images. The preparation process was similar to the one I explained here: http://www.photomacrography.net/forum/viewtopic.php?t=19666&highlight=electron

Best,
Ecki
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