Vibrant DIC videos

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microscopeboi
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Vibrant DIC videos

#1 Post by microscopeboi » Wed Mar 16, 2022 1:11 pm

Hello,

Are the DIC videos on youtube edited to add color?

I assume they have to be because DIC is in black and white usually right?

If so, whats a good software for this?

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Re: Vibrant DIC videos

#2 Post by zzffnn » Wed Mar 16, 2022 2:11 pm

Please read on DIC first, then put forward an assumption, if you have to. Hint: DIC color is usually grey, blue or pink, sometimes dark navy blue or almost black and white, but usually not black and white, unless you purposely set it that way.

What I learned from graduate school is, always assume I don’t “know enough”; if I have to put forward a HYPOTHESIS, be ready to accept that it is wrong.

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Re: Vibrant DIC videos

#3 Post by Microscopy_is_fun » Wed Mar 16, 2022 7:23 pm

microscopeboi wrote:
Wed Mar 16, 2022 1:11 pm
If so, whats a good software for this?
Hi,

the vibrant colours you see in many DIC images are typically achieved using the right "hardware".

When utilizing the DIC equipment, you will usually start by setting the prisms to a neutral position. This is done by first putting the two polarizers to a position where all light is blocked. Then you will move in one DIC prism and take off the eyepiece. You will then see some kind of rainbow, and in the initial position the black bar is in the center. The same is done with the second prism.

Now you can plug in in the eyepiece and put in the sample. By turning the screw on one of the prisms, the "rainbow" will move, and in further consequence the contrast changes and the background turns from light to dark. Depending on your sample, you will also see some colour changes, but basically the image is black and white in case of uncoloured samples (e.g. many ciliates).

To get the "popping" colours which DIC is known for, you need a lambda-plate (in my case, this is a specific polymer film, see image) which is placed between polarizer and the condensor prism. By rotating the lambda-plate you will provoke the psychedelic colours, which you are probably referring to.

Software is not necessary for that.
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Re: Vibrant DIC videos

#4 Post by Phill Brown » Wed Mar 16, 2022 9:38 pm

Then there is the digital imaging system connected to the computer which runs graphics drivers and image enhancement software.

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Re: Vibrant DIC videos

#5 Post by Microscopy_is_fun » Wed Mar 16, 2022 10:34 pm

Phill Brown wrote:
Wed Mar 16, 2022 9:38 pm
Then there is the digital imaging system connected to the computer which runs graphics drivers and image enhancement software.
You are right, image processing certainly has a large effect on the finally obtained colours. And not to forget the camera, which (in my case) has quite some trouble with DIC-images. I guess the histograms of the RGB-channels differ quite a lot from conventional photographs. Thus the camera software wants to bring in some more colour into the mostly greyish DIC images, easily yielding oversaturation of the recorded image.

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Re: Vibrant DIC videos

#6 Post by Tom Jones » Wed Mar 16, 2022 11:23 pm

microscopeboi,

For a little bit of introductory homework, see:

https://en.wikipedia.org/wiki/Different ... microscopy

https://www.olympus-lifescience.com/en/ ... /dicintro/

https://en.wikipedia.org/wiki/Waveplate

https://www.youtube.com/watch?v=FUa1GTc69y4

Colors, including on some systems a near darkfield black background, will be determined by the system (Zeiss, Olympus, etc., will not be the same on any given specimen as the prism shear values are different), the amount of additional shear applied to the second prism by translation, the specimen, the rotation of the specimen in the light path (remember, it's polarized light with extra prisms), and the presence or absence of any given wavelength waveplate, or slightly misaligning the polarizers.

It is a very complex and expensive research method, and the colors produced are real, just not particularly relevant. Post processing is quite unnecessary for color DIC, and much more difficult in video than stills.

Tom

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Re: Vibrant DIC videos

#7 Post by Scarodactyl » Wed Mar 16, 2022 11:50 pm

My Nikon E800 does not produce color by default except with the 100x prism which shows some when pulled out of position. I think it would need the wave plate to do so.

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Re: Vibrant DIC videos

#8 Post by Tom Jones » Thu Mar 17, 2022 12:04 am

My Olympus BHS system produces nice dark backgrounds as well as the full range of colors depending on how far the analyzer prism is translated. On the other hand, my Olympus BX system with the DICT slider will not produce a good darkfield effect with or without the 530nm waveplate. Only the full range of colors with prism translation. I can get the dark grey background with the DICTS slider.

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Re: Vibrant DIC videos

#9 Post by Microscopy_is_fun » Thu Mar 17, 2022 8:44 pm

Tom Jones wrote:
Wed Mar 16, 2022 11:23 pm
..., and much more difficult in video than stills.
A quite useful and professional video-editing tool, which is available for free and provides powerful color correction tools is Davinci Resolve. I am rather new to editing of microscope-videos, and certainly use less than 10% of Davinci's capabilities, but it seems to work quite well for footage taken with DIC


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Re: Vibrant DIC videos

#10 Post by josmann » Tue May 17, 2022 1:25 am

Microscopy_is_fun wrote:
Thu Mar 17, 2022 8:44 pm
A quite useful and professional video-editing tool, which is available for free and provides powerful color correction tools is Davinci Resolve. I am rather new to editing of microscope-videos, and certainly use less than 10% of Davinci's capabilities, but it seems to work quite well for footage taken with DIC
One trick I can share about Davinci is to increase the parameter called Mid/Detail on the color wheels tool. It sharpens the image by some form of magic that doesn't cause it to look oversharpened.

I use Davinci for all my video work and to generate LUTs for my live streamed DIC footage.

Here's a clip of me adjusting my DIC prism translation to generate the fun colors: https://www.twitch.tv/diettoms/clip/Bor ... lsggcdUwu7
Live DIC microscopy T/TH/SU at 8PST: https://www.twitch.tv/diettoms

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Re: Vibrant DIC videos

#11 Post by rogeliomoreno » Fri Aug 19, 2022 5:15 am

Scarodactyl wrote:
Wed Mar 16, 2022 11:50 pm
My Nikon E800 does not produce color by default except with the 100x prism which shows some when pulled out of position. I think it would need the wave plate to do so.
With my Nikon TE300 to produce background color you need to pull out the objective's prism (around 3 mm) then you rotated the polarizer to change the background color, no need of the lambda plate. I am using the same DIC system that use condenser's prisms N1, N2; I also have the old DIC system that use L, M condenser's prisms and can get background color with the same method. You can also get background color using the lambda plate and rotating the polarizer (in this case you leave the objective's prism full inserted).

Rogelio

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Re: Vibrant DIC videos

#12 Post by Macro_Cosmos » Thu Nov 10, 2022 12:24 am

I cannot really fault the OP for making the false assumption. I believe Nikon and Zeiss DIC is basically monochromatic with the nosepiece prism slid all the way in. Pulling the prism out slightly will generate the "colours".
There are two mechanical implementations of DIC from the big 4 players with different characteristics. Zeiss and Nikon uses small nosepiece prisms for each objective, Leica and Olympus uses a common nosepiece slider which allows the prism to be translated precisely (the type I prefer for my work).

Assuming the normal DIC is being used.
Leica: higher contrast, common nosepiece slider with condenser prisms for specific objetives.
Nikon and Zeiss: contrast and resolution balanced, most people prefer the look of these two. Each objective has its own slider while condenser prisms are tailored for different NA.
Olympus: Between Leica and Nikon/Zeiss. Common nosepiece slider with condenser prisms for specific objectives. High contrast at lower magnifications while contrast and resolution is balanced at higher magnifications.

Generally, I am of the opinion that Nikon/Zeiss is the "best" for video since contrast and resolution is very well balanced. Leica and Olympus produces way too much contrast at lower magnifications and the variation is huge.

Here is a visualisation using the best software I have access to:
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"Normal" DIC only, the big 4 also offers high contrast and high resolution variants and those are very different. For example, Olympus' HR type at 10x is basically the same as Nikon's normal type. This is why many would claim Zeiss and Nikon DIC has higher resolution. The claim is technically true but it really is about the manufacturer's vision and how they balance between contrast and resolution, and therefore the type of sample (too thick or thin = destructive glare with normal DIC!). :lol:

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Re: Vibrant DIC videos

#13 Post by Sure Squintsalot » Thu Nov 10, 2022 4:40 am

"pulling the prisms out will generate the colors"

Edited: pulling one of the prisms partially out prevents sheared light waves from being properly realigned. This generates the wild colors, at least, in the case of my static images of crystal surfaces.

It really is too bad that the colors are relatively meaningless, despite the fact that they can mesmerize me for hours.

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Re: Vibrant DIC videos

#14 Post by Macro_Cosmos » Sat Nov 12, 2022 12:54 pm

Sure Squintsalot wrote:
Thu Nov 10, 2022 4:40 am
"pulling the prisms out will generate the colors"

Edited: pulling one of the prisms partially out prevents sheared light waves from being properly realigned. This generates the wild colors, at least, in the case of my static images of crystal surfaces.

It really is too bad that the colors are relatively meaningless, despite the fact that they can mesmerize me for hours.
So basically, both Nomarski prisms are fixed on the current iteration of the Nikon system? I think the tiny Zeiss sliders can be adjusted though.
A full-wave retarder can be used I suppose, those colours are lovely.
Otherwise, a de Sénarmont compensator can be inserted and the analyser can be rotated to bring in bias.

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Re: Vibrant DIC videos

#15 Post by PeteM » Sat Nov 12, 2022 4:32 pm

Macro_Cosmos wrote:
Sat Nov 12, 2022 12:54 pm
. . . So basically, both Nomarski prisms are fixed on the current iteration of the Nikon system? . . .
Nikon Eclipse systems use individual prisms on sliders above each objective for their infinity systems, much like Zeiss. They are also commonly equipped with a wave plate. So, there is a lot of flexibility.

My system (an ME600) is the earlier type with three prisms in the condenser - low, medium, and high. The latest Nikon infinity system gets by with two prisms in the condenser and then a dizzying and costly array of individual objectives and prisms.

In the finite world, Nikon made systems with fixed condenser prisms and a slider above (much like Olympus finite and infinite) AND a system much more rarely seen with individual prisms above each objective. Those prisms are also on small sliders, with some adjustment.

Olympus also made different infinity sliders - one is normal, one is aimed at the higher resolution, and the third at higher contrast.

Personally, I'm a fan of the Leica DIC. It can achieve very high contrast and a stunning false "3D." Combinations of prisms are available to get some meaningful DIC effect all the way down to a 2.5x objective and stunning DIC effects from 5x to 60 or 100x 1.4na. Still, any time I look at Nikon, Olympus, or Zeiss - those immediately become new favorities.

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Re: Vibrant DIC videos

#16 Post by Macro_Cosmos » Mon Nov 14, 2022 9:54 pm

PeteM wrote:
Sat Nov 12, 2022 4:32 pm
Macro_Cosmos wrote:
Sat Nov 12, 2022 12:54 pm
. . . So basically, both Nomarski prisms are fixed on the current iteration of the Nikon system? . . .
Nikon Eclipse systems use individual prisms on sliders above each objective for their infinity systems, much like Zeiss. They are also commonly equipped with a wave plate. So, there is a lot of flexibility.

My system (an ME600) is the earlier type with three prisms in the condenser - low, medium, and high. The latest Nikon infinity system gets by with two prisms in the condenser and then a dizzying and costly array of individual objectives and prisms.

In the finite world, Nikon made systems with fixed condenser prisms and a slider above (much like Olympus finite and infinite) AND a system much more rarely seen with individual prisms above each objective. Those prisms are also on small sliders, with some adjustment.

Olympus also made different infinity sliders - one is normal, one is aimed at the higher resolution, and the third at higher contrast.

Personally, I'm a fan of the Leica DIC. It can achieve very high contrast and a stunning false "3D." Combinations of prisms are available to get some meaningful DIC effect all the way down to a 2.5x objective and stunning DIC effects from 5x to 60 or 100x 1.4na. Still, any time I look at Nikon, Olympus, or Zeiss - those immediately become new favorities.
Thanks for the information. I have only used Nikon and Zeiss in the lab.
I can get decent DIC with my Olympus 4x by using a 25x prism and fiddling with the BFP1 slider.
I am personally not a fan high contrast, that is something I am happy to add or subtract during the post processing stage. Both Nikon and Zeiss has/had high contrast and high resolution types as well. Nikon even went further with their DIC-SS which offered higher resolution than HR, though only for E-series (my bad!).
(SS means short shear, this was used for VEC = video enhanced contrast).
Last edited by Macro_Cosmos on Thu Nov 17, 2022 12:55 am, edited 1 time in total.

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Re: Vibrant DIC videos

#17 Post by Scarodactyl » Mon Nov 14, 2022 10:12 pm

Nikon made ss prisms for the eclipse.

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Re: Vibrant DIC videos

#18 Post by Macro_Cosmos » Thu Nov 17, 2022 12:55 am

Scarodactyl wrote:
Mon Nov 14, 2022 10:12 pm
Nikon made ss prisms for the eclipse.
Thanks, I messed up and did not read the graph I had properly. :roll:

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