Luminous Landscape Forum

The Art of Photography => User Critiques => Topic started by: PSA DC-9-30 on August 09, 2012, 03:14:19 pm

Title: Microbe cover (December 2011)
Post by: PSA DC-9-30 on August 09, 2012, 03:14:19 pm
I am a biologist and imaging specialist, and have developed new techniques for imaging microbes (especially protozoa) with scanning electron microscopy. My images have been featured on seven journal/magazine covers, textbooks, newspapers, etc.

This is a recent one I wanted to share. This image was taken while I was a postdoc at the University of British Columbia.

(http://www.microbemagazine.org/images/stories/dec2011/webcover.jpg)

Caption: Scanning electron micrograph of an anaerobic parabasalid flagellate protist (about 40 um long) from the hindgut of the termite Neotermes castaneus covered with elongate bacterial symbionts. Protists are globally significant but often overlooked microbes (see p. 516). (Image © Kevin Carpenter and Patrick Keeling, Lawrence Livermore National Laboratory and University of British Columbia.)

http://www.microbemagazine.org/index.php/issue-archive/2011-issues/december-2011

Title: Re: Microbe cover (December 2011)
Post by: Richowens on August 09, 2012, 05:46:42 pm
Congratulations Kevin.

Rich
Title: Re: Microbe cover (December 2011)
Post by: Eric Myrvaagnes on August 09, 2012, 07:43:18 pm
Stunning shot! Congrats!
Title: Re: Microbe cover (December 2011)
Post by: WalterEG on August 09, 2012, 07:56:05 pm
Well deserved recognition right there.
Title: Re: Microbe cover (December 2011)
Post by: RSL on August 09, 2012, 09:09:01 pm
Fascinating stuff.
Title: Re: Microbe cover (December 2011)
Post by: Slobodan Blagojevic on August 09, 2012, 11:16:16 pm
Simply beautiful! And beautifully simple. A cross between a fashion shot and fine art, let alone science.
Title: Re: Microbe cover (December 2011)
Post by: PSA DC-9-30 on August 10, 2012, 12:03:28 am
Thanks very much for the kind words. I will be posting some other covers and projects soon. And one really big, exciting announcement for April 2013.

In the mean time, I must be the only photographer here using a Hitachi "camera"  ;D

(http://electronmicroscopy.org/images/FE_SEM_Microscope.jpg)
Title: Re: Microbe cover (December 2011)
Post by: wolfnowl on August 10, 2012, 02:21:23 am
In the (many) years I worked with the rabies unit we had a Zeiss scope with a camera for UV and phase shift microscopy, but nothing quite so fancy as that!

Mike.

P.S. Congratulations well deserved.
Title: Re: Microbe cover (December 2011)
Post by: Eric Myrvaagnes on August 10, 2012, 08:54:08 am
I'd like to hear Kent Rockwell's ("Your camera doesn't matter") response to this image and your camera!  :D
I'll readily admit I couldn't have done it with my Canon 5D2.
Title: Re: Microbe cover (December 2011)
Post by: francois on August 10, 2012, 11:30:38 am
Late congrats... Beautiful image!
Well done.
Title: Re: Microbe cover (December 2011)
Post by: Justan on August 10, 2012, 11:35:02 am
How does the camera get the dof? Is this the result of multiple images and focus stacking or ??
Title: Re: Microbe cover (December 2011)
Post by: PSA DC-9-30 on August 10, 2012, 10:50:18 pm
Greater DOF with secondary electron imaging is largely a matter of working distance--defined as the distance (in mm) from the objective lens to the top of the sample being imaged. Of course, the lenses in this type of instrument are electromagnetic (not glass) lenses, and can effect different crossover (focus) points based on current supplied to the lens coils.

 The longer the WD, the greater the DOF, (but this entails other tradeoffs as with every operating parameter). Of course, this is a familiar principle to any photographer; the closer you move to an object, the shallower the DOF is.

The WD I used for this shot was 28mm, which is considered very long. I also use a tilt of around 30 degrees. This adds an additional sense of depth. If you were trying to convey the three dimensionality of a sphere, or ping pong ball for example, the worst way to photograph it would be from directly above. Better to come in obliquely from the side.  

The protozoa (protists is a better word) that live in the guts of lower termites are often very large, and this presents a challenge for DOF. This one in question is about 40 microns long, but others can be up to 300 microns long. We beleive that they have evolved large size in order to engulf the relatively large wood fragments that make their way to the hindgut after being chewed by the termites jaws.

Focus stacking is something I've never tried, but for some large cells, I've taken multiple images with different portions in focus. If someone can point me to a tutorial for focus stacking, in Photoshop (I use CS2) I would appreciate it!
Title: Re: Microbe cover (December 2011)
Post by: Dave (Isle of Skye) on August 12, 2012, 12:56:37 pm
Kevin,

I don't think you can do focus stacking in CS2 if memory serves me correctly, I think it came along in CS3 and onwards. But you can go here (http://www.heliconsoft.com/heliconfocus.html) for the software that most people seem to prefer, although I do not use it myslef, so cannot comment on how good it is.

Fantastic shot by the way, so how much does that setup cost and where would I go to buy one??? ;D

Dave
Title: Re: Microbe cover (December 2011)
Post by: RSL on August 12, 2012, 05:07:25 pm
In the mean time, I must be the only photographer here using a Hitachi "camera"  ;D

Is there an Upstrap available for this camera?
Title: Re: Microbe cover (December 2011)
Post by: Eric Myrvaagnes on August 12, 2012, 05:52:13 pm
I bet it doesn't even have a mirror lockup button.  ;)
Title: Re: Microbe cover (December 2011)
Post by: Justan on August 13, 2012, 01:12:33 pm
Greater DOF with secondary electron imaging is largely a matter of working distance--defined as the distance (in mm) from the objective lens to the top of the sample being imaged. Of course, the lenses in this type of instrument are electromagnetic (not glass) lenses, and can effect different crossover (focus) points based on current supplied to the lens coils.

 The longer the WD, the greater the DOF, (but this entails other tradeoffs as with every operating parameter). Of course, this is a familiar principle to any photographer; the closer you move to an object, the shallower the DOF is.

The WD I used for this shot was 28mm, which is considered very long. I also use a tilt of around 30 degrees. This adds an additional sense of depth. If you were trying to convey the three dimensionality of a sphere, or ping pong ball for example, the worst way to photograph it would be from directly above. Better to come in obliquely from the side.  

The protozoa (protists is a better word) that live in the guts of lower termites are often very large, and this presents a challenge for DOF. This one in question is about 40 microns long, but others can be up to 300 microns long. We beleive that they have evolved large size in order to engulf the relatively large wood fragments that make their way to the hindgut after being chewed by the termites jaws.

Focus stacking is something I've never tried, but for some large cells, I've taken multiple images with different portions in focus. If someone can point me to a tutorial for focus stacking, in Photoshop (I use CS2) I would appreciate it!


Thanks very much for this explanation. Never worked with or read about this kind of equipment before so I still didn’t quite get it. Due to that I asked to Mr. Google who provided the following reference. Of course there could be a lot of different SEMs but the key details that caught my eye were:

“The scanning electron microscope has many advantages over traditional microscopes. The SEM has a large depth of field, which allows more of a specimen to be in focus at one time.” http://www.purdue.edu/rem/rs/sem.htm#2

My next question is: does a SEM require a light source? The shading on the image you displayed is so delicate it made me wonder how one could position one or more lights to produce the result on such a small object.

Quote
We believe that they have evolved large size in order to engulf the relatively large wood fragments that make their way to the hindgut after being chewed by the termites jaws.

Any time a case for selection can be exemplified that is pretty cool! I guess it’s a completely different discussion but I have to ask:  Are only the larger protists found in adult termites? Do they grow in size as the termite does?

This is all way outside of my experience, so I hope you don’t mind some questions.

WRT focus stacking, there are a few here who have a lot of experience with focus stacking software. I think that kind of technology might be very useful for this kind of work.

Remember us when you get a Nobel for your future work! <big toothy grin>
Title: Re: Microbe cover (December 2011)
Post by: sailronin on August 13, 2012, 01:26:28 pm
Stunning image and great explanation in simple terms of the setup.
Really interesting to see a different type of imaging presented.
Title: Re: Microbe cover (December 2011)
Post by: cmi on August 16, 2012, 07:24:49 pm
Thanks very much for the kind words. I will be posting some other covers and projects soon. And one really big, exciting announcement for April 2013.

In the mean time, I must be the only photographer here using a Hitachi "camera"  ;D

(http://electronmicroscopy.org/images/FE_SEM_Microscope.jpg)

Why dont you write up an article for Lula. That would fit nicely with the Space and Astro-Landscape Photography articles.
Title: Re: Microbe cover (December 2011)
Post by: Eric Myrvaagnes on August 16, 2012, 11:24:09 pm
Why dont you write up an article for Lula. That would fit nicely with the Space and Astro-Landscape Photography articles.

Second the motion!
Title: Re: Microbe cover (December 2011)
Post by: Tony Jay on August 17, 2012, 01:06:14 am
I did both transmission and scanning electron micrography in the late 1980's while still at university.
It was a lot of fun.

It would really be very interesting to see what has changed.
So, I would add my support to the suggestion that you write an article for LuLa about electron micrography.

Regards

Tony Jay
Title: Re: Microbe cover (December 2011)
Post by: PSA DC-9-30 on August 17, 2012, 12:14:42 pm
Hi,

Have been meaning to post more about the biology of these organisms, but I have been busy and tired these days.

What is LuLa? Luminous Landscape? But what section?

I am also interested in going on Science Friday and will apply to TED one of these days. My research seminars and talks always go over quite well, and people love the images. I think people have very little understanding of what protists actually are and why they are so important in many different respects.

Does anyone have any ideas on how I might get publicity for the images, and what galleries may be interested in this type of work? (will post more photos later).

Thanks
Kevin
Title: Re: Microbe cover (December 2011)
Post by: Tony Jay on August 17, 2012, 07:08:33 pm
Yes Kevin, LuLa is shorthand for Luminous Landscape.

Hopefully we are getting Michael Reichman's attention and the idea is that he might invite you to submit an article on the subject of electron micrography. If it were to happen it would not be posted on the forum but published onto the main site.

Regards

Tony Jay
Title: Re: Microbe cover (December 2011)
Post by: PSA DC-9-30 on August 18, 2012, 01:15:55 am
Hi Tony,

Well if someone wants to bring this to Michael's attention, I would be glad to do it.

Kevin