Composite marsh organisms and flatworm

Images made through a microscope. All subject types.

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Linden.g
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Composite marsh organisms and flatworm

Post by Linden.g »

The first image is a composite of organisms found in a fresh water marsh. One of the most complex ecosystems I've come across to date. Dominated by dismids rather than diatoms. DIC flash, Olympus 100x achromat. Second image from a spring, large ciliate with injested rotifer. DIC flash, Olympus 20x achromat.
Image
Image
Linden
Last edited by Linden.g on Mon Apr 04, 2011 11:37 am, edited 1 time in total.

Franz Neidl
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Post by Franz Neidl »

Hello Linden,

I like very much your pictures! Congratulations.
In the second picture is not a ciliate, but a flatworm.
A question to the first picture: How do you make a composition? With photoshop?

Franz

Mitch640
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Post by Mitch640 »

Beautiful images. I think the worms name must be Casper. :)

Linden.g
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Post by Linden.g »

Franz, thanks for the correction on the ID. With respect to the composite, these are the steps:
1. Once your happy with a sample slide and decide to make a composite, you tune the DIC and then don’t adjust again until all images have been collected.

2. Batch process raw images in Photoshop CS4, critical to treat them all the same way

a. adjust colour balance to more neutral gray via temperature

b. center the exposure

c. Reduce chromatic aberration via adjustment of saturation of HSL

d. Reduce noise

e. Save files as TIF

3. For organisms which you plan to do a focus stack, stack in Zerene stacker and save as TIF

4. Open all TIF images in PS CS4

5. Choice one image which has the smallest organism and duplicate the background layer to make a new layer

6. Blur the new layer with Gaussian blur to make a smooth even layer which you will use as a mask

7. Cut each organism from the individual images and past as new layers on the first image. No need to be careful with cutting just use square sections. Move them around to make a pleasant composition.

8. Move the blurred layer to the top and make it 50% intensity. You will then see all of the other organisms through this layer

9. Using the eraser tool, erase the top layer in the areas where you see the organisms below, this makes windows in the top layer and allows the organisms below to show through.

10. Make the top layer 100% intensity and this then gives a the whole stack a smooth background.

11. Flatten the layers

12. Adjust colour balance, contrast etc, via curves, levels

13. Reduce image size, sharpen, convert to sRBG colour space and change to 8 bit, save as JPEG

Hope that helps

Linden

Franz Neidl
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Post by Franz Neidl »

Thank you, Linden, for your detailed answer. Now I know clearer that I still need to learn a lot!

Franz

Linden.g
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Post by Linden.g »

Hi Frans, at least you can identify organisms :oops: :) , If you need any help, let me know.
Linden

Charles Krebs
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Post by Charles Krebs »

Very nicely done Linden. Excellent work!

Cactusdave
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Post by Cactusdave »

Linden.g wrote:
a. adjust colour balance to more neutral gray via temperature

c. Reduce chromatic aberration via adjustment of saturation of HSL
I really appreciate you detailing your workflow for this neat compositing technique. Can I ask you to give a bit more information on these two steps please.
Leitz Ortholux 1, Zeiss standard, Nikon Diaphot inverted, Canon photographic gear

Linden.g
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Post by Linden.g »

Hi Dave, here is a little more detail

a. adjust colour balance to more neutral gray via temperature

My DIC gives a strong blue colour so during RAW conversion I use the "temperature" slider which is calibrated in Kelvin's to reduce this. Incandescent light is a around 3500K the flash and DIC gives a colour bias as high as 10,000K http://www.3drender.com/glossary/colortemp.htm . You have to be careful not to over do it or you will loose reds, yellows and greens. I also tweak the "tint" to compensate for any off colour. You have another chance to reduce the blue with the colour saturation sliders once you are in PS4.

c. Reduce chromatic aberration via adjustment of saturation of HSL

The RAW converter also has chromatic aberration slides which really work well with standard photographic lens. I haven't found them sufficient for my S-Plan Achromats. So I aften resort to the "Hue, Saturation, Luminance" controls in the RAW converter. By using the saturation slides you can reduce the aberration due to magenta, purple, blues and Aquas. This also make the background closer to gray. As most organisms don't show these colours this wont take wanted detail out of the subject. Again take care that this doesn't remove detail you want to keep.

Hope that helps

Linden

Cactusdave
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Post by Cactusdave »

Thanks very much Linden. The tip about reduction of chromatic aberration during RAW conversion is especially helpful.
Leitz Ortholux 1, Zeiss standard, Nikon Diaphot inverted, Canon photographic gear

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