Nikon E600/E800/E1000/TE200/TE300 vs 80i/90i/Ti DIC system

RogelioMoreno · Post by **RogelioMoreno** » Sat Jan 28, 2012 1:20 pm

Here are the results of the test.

The first two pictures were taken with the 80i/90i/Ti DIC system, the last two with the E600/E800/E1000/TE200/TE300 DIC system.

All the images with Nikon TE300 microscope, Nikon Plan Apo 20x/0.75 objective, DIC (for the 80i... system: N2 condenser prism with the 20x objective prism, for the E600... system: M condenser prism with PF 20x objective prism), flash and Canon T3i mounted on the front port of the TE300. All crop from the full frame.

I do not see too much differences.

Rogelio

Will Milne · Post by **Will Milne** » Sat Jan 28, 2012 6:00 pm

Hi

You make some of the most compelling and beautifull scope images I have ever seen . Both you and Charles K have demonstrated that DIC is a very seductive lighting option .

I am sure I am not alone in pondering /wishing for a DIC rig for the wonderfull color/tonal/detail imagery it seems to produce in skilled hands . .

Despite it's obvious and undeniable aesthetic appeal I am curious about the information that suggests that the results from DIC are tempered by the manner in which DIC creates the appearance of visual information. In particular the the concept that some of the information in a DIC image is a result of the approach itself.

I was curious if anyone has compared images done of the same subject at the same magnifications using say oblique lighting ( or some other contrast enhancing approach)compared to DIC and looked at the results without consideration for aesthetics.

Maybe the question I am asking in my usual roundabout way- why would you use DIC and why would you not instead use another contrast enhancing approach????????? What does DIC reveal compared to other approaches and what does it obscure????

Will

Litonotus · Post by **Litonotus** » Sun Jan 29, 2012 1:50 am

I see no difference here. maybe a difference is noticable when the high contrast or high resolution modules from the i system are used. by the way - did you try to mix those systems...?

both are wonderful.

using say oblique lighting compared to DIC and looked at the results without consideration for aesthetics

that would be extremely hard

Cactusdave · Post by **Cactusdave** » Sun Jan 29, 2012 2:01 am

The evenness and colour depth of you DIC background is compelling, as well as the sharpness of the image. I assume all your backgrounds are 'as shot'? To get anything like that background I have to play around in Photoshop, to the detriment sometimes of the main image.

RogelioMoreno · Post by **RogelioMoreno** » Sun Jan 29, 2012 4:31 am

Litonotus wrote:I see no difference here. maybe a difference is noticable when the high contrast or high resolution modules from the i system are used. by the way - did you try to mix those systems...?

I do not have the high contrast neither the high resolution modules from the i system, so no test can be done. I did try to mix the E600... with the 80i... DIC and that did not work.

Rogelio

RogelioMoreno · Post by **RogelioMoreno** » Sun Jan 29, 2012 4:39 am

Cactusdave wrote:The evenness and colour depth of you DIC background is compelling, as well as the sharpness of the image. I assume all your backgrounds are 'as shot'? To get anything like that background I have to play around in Photoshop, to the detriment sometimes of the main image.

The as shot background is more blue, I had to use a color cast correction to remove some amount of blue from the pictures. These are crop and the TE300's front port has a 2x photo lens so the images a little part of the image generated by the objective, I will try to post full frame images (using the 2x of the front port) so you can compare background evenness.

Rogelio

RogelioMoreno · Post by **RogelioMoreno** » Sun Jan 29, 2012 5:20 am

Will,

Check the following link for comparison of Phase Contrast and DIC:

http://micro.magnet.fsu.edu/primer/tech ... rison.html

Thank you for your comments.

Rogelio

RogelioMoreno · Post by **RogelioMoreno** » Sun Jan 29, 2012 5:55 am

Here full frame from both systems, picture 1 with 80i... and picture 2 with E600... DIC system.

I only applied: contrast adjustment, color cast correction, sharpenning and noise reduction.

Rogelio

Cactusdave · Post by **Cactusdave** » Sun Jan 29, 2012 7:29 am

Thanks Rogelio. That's very impressive.

Litonotus · Post by **Litonotus** » Sun Jan 29, 2012 11:32 am

I did try to mix the E600... with the 80i... DIC and that did not work.

one of important for me questions is answered, thank you (:

discomorphella · Post by **discomorphella** » Sun Jan 29, 2012 1:37 pm

Hi Rogelio,

Your objective / scope / camera combination is really very sharp, and I don't think your protozoan sample is really testing its ultimate resolution. If you can find something like a Cymbella or similar diatom, which has a pore pitch of ~0.5 micron, then we could see the contrast difference in the pores for each optical condition, or choice of DIC equipment etc. At the diffraction limit, your 0.75 NA objective should be capable of resolving the 0.5 micron pore pattern at approximately 0.25 contrast, assuming all other sources of aberration etc are absent and you have an adequate number of pixels per Airy spot etc.

For Will, DIC has several advantages over oblique (off-axis) illumination or phase contrast (which also has off-axis illumination in its setup). Among the most significant advantages, it is less sensitive to out-of-focus light. The gives you increased contrast for the in-focus part of the image (the part we care about). Because it allows you to get decent contrast with more of the objective back plane illuminated (less spatial coherence) a DIC image is less degraded by out-of-plane light. I should qualify this because the actual transfer function for a DIC scope is quite a bit more complex than for a regular scope with off-axis illumination and the things that degrade a DIC image are not the same as for simpler optics. However, since a lot of the things we like to observe, particularly live, aquatic specimens, are more than a few focal planes thick for a higher NA (> 0.65) objective, DIC has an advantage over phase or oblique. On the other hand, for very thin specimens, e.g., a flagellum on a Euglena or similar, phase may give better contrast. As you try and focus through the entire animal, you'll notice that the phase image will be degraded by information from above/below the focal plane. Hopefully this wasn't too confusing.

David

RogelioMoreno · Post by **RogelioMoreno** » Sun Jan 29, 2012 2:05 pm

David,

I have a slide with Lyrella lyra, would that diatom ok for tthe test?

Rogelio

discomorphella · Post by **discomorphella** » Sun Jan 29, 2012 2:13 pm

Hi Rogelio,

I am not sure of what the pore pitch is for that particular diatom, but it should be sufficiently less than 1 micron to give a good test. The main advantage of the diatom is that it will be easier to focus on the exact same structure each time. This will ensure that you can test your combinations of DIC optics consistently. The protozoa look nice, but they are hard to maintain in the exact same position and orientation. The orientation of the various structures with respect to the optical axes of your prisms also affects the DIC resolution.

David