DIC versus other forms of Microscopy

Images made through a microscope. All subject types.

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Robert Berdan
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DIC versus other forms of Microscopy

Post by Robert Berdan »

I have just posted a new article on my website about DIC microscopy and how it compares to other microscopy methods.
https://www.canadiannaturephotographer. ... scopy.html

I have also been trying to find software previously described that simulates DIC and Phase contrast images from focus stacking three bright field images described by B.E. Allman et al 2002. Optical Phase Microscopy: Quantitative Imaging and Conventional Phase Analogs 2002. Microscopy and Analysis. 13-15
https://www.researchgate.net/publicatio ... se_Analogs

Anyone with information about the software or similar software I would appreciate learning about it.

Below are a couple of images from my article about DIC that I think might of interest to anyone interested in DIC imaging.
Attachments
Amoeba DIC 200X
Amoeba DIC 200X
Ciliate Phacodinium found in Moss DIC
Ciliate Phacodinium found in Moss DIC
Air Bladders DIC Chaoborus midge fly larva
Air Bladders DIC Chaoborus midge fly larva
Peritich Ciliate 400X
Peritich Ciliate 400X

Scarodactyl
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Re: DIC versus other forms of Microscopy

Post by Scarodactyl »

That was a nice read and the images are fantastic.
I wonder if Sanderson DIC is going to end up being a big deal. I've only seen Saul's sample images but they looked extremely promising, and it should be much cheaper than Nomarski.
Epi DIC systems can be surprisingly cheap, and from time to time I wonder if you could pair them with a polarizer and slit/prism below the condenser and have a chance at getting transmitted DIC. I'd guess it would at least be pretty hard, since transmitted DIC is so fiddly (vs episcopic where I seem to have a decent chance at getting good DIC on my scope even with random lenses).

Lou Jost
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Re: DIC versus other forms of Microscopy

Post by Lou Jost »

Yes, a very interesting article. Thanks for writing and sharing it.

I think you may have been overly pessimistic about the cost of fluorescent microscopy. Secondhand fluorescence epi-illuminators are often available on eBay for older microscopes (e g Optiphot) for around $200. Bright UV LEDs are less than $100. Filter cubes might cost a few hundred dollars.
And you can get by without most of that, by using a stripped-down normal epi-illuminator and a UV flashlight and some filters. Even a dichroic mirror is not absolutely necessary. Indeed, even an epi-illuminator is not strictly necessary for low magnifications.

I also think that the scope of fluorescent microscopy is not limited to specimens treated with fluorescent marker dyes. I think for non-research purposes, natural fluorescence is much more interesting and is practically cost-free. And unlike most of the other techniques you mention, fluroescence photography is feasible at even very low m, including the ordinary macro realm.

Robert Berdan
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Re: DIC versus other forms of Microscopy

Post by Robert Berdan »

Hi Lou - I understand one can even make their own Fluorescence microscope, they still require dichroic filters etc and the cost of new Fluorescence attachment rrom China are in the order of $5,000 e.g. visit Motic.com web site - sometimes you can get lucky and get lower cost scopes or irridiate t0eh specimens directly with UV lights like from Night Sea. My used Nikon Fluorescent scope cost $600 online, but after adding a new bulb, etc and and proper high NA objectives etc it's cost is now closer to $6,000. I wish I could find an LED fluorescent attachment for Axioscope but no luck so far and most commercial light sources I have inquired about especially LED are several thousand - hopefully this will get lower in the future. Zeiss has a very nice LED light source for $11,000 but out of my price range for now. Also I have read tutorials for teachers on how to make a Fluorescent microscope using LED flash lights - so your comments are correct for those that can build their own microscope with cheaper components.

I do use my flourescence microscopes with autoflourescence subjects like minerals, algae - e.g. spirogyra etc , and lichens etc that are autofluorescent . I have photographed these things and I also have a UV light for my stereoscope for looking for fluorescent minerals and lichens etc. My comments are meant as a "generalization" about the costs of new fluorescent microscope compared to a bright field, or phase scope in general .

I have not found a DIC microscope for less then $10, 0000 even a used microscope (not including epi-DIC) scopes. I get a lot of questions folks from folks wanting DIC until they find out how expensive it is. I am hoping software will reduce the price. Many of us make or refurbish older microscopes from parts and that helps. DIC isn't something that is easy to make at least I have never seen it and many Chinese microscope companies don't offer them maybe because of patents or they are just difficult to make I am not sure.

Thank you kindly for you comments my goal is to promote microscopy and simply show different modes of lighting generally available with a rough idea of the costs for each type - I think a Rheinberg filter set for $40 on E-bay is one of the best deals out there and their many different types of microscopes and approaches to microscopy. Most folks I talk to want to own a DIC scope before they have ever used a basic scope and I am trying to point them toward phase contrast or bright field microscopy first and then add Rheinberg or polarizing filters.
Thank you Lou I appreciate your comments - and agree with you.
RB
Last edited by Robert Berdan on Fri May 14, 2021 6:42 pm, edited 1 time in total.

Robert Berdan
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Re: DIC versus other forms of Microscopy

Post by Robert Berdan »

Hi Scarodactyl - I looked at some articles about Sanderson contrast technique sometime ago, but I wasn't that impressed with the images and ended up filing the paper somewhere. I am hoping that DIC and phase contrast will eventually be available to everyone by using software to process bright field images. Phase is not that expensive. I had to wait a long time before I could purchase a DIC scope for home use. I searched EBay and online for a used one. Finally found one from Vermont Optical for $10,000 - it was on an older Olympus DIC scope and said what the hell I going to buy a new scope before I die - at my age I deserve it. Instead of a new car I bought a new microscope :-) so that is what I say I drive these days is a Zeiss Axioscope and it is wonderful. Also had to sell most of my other toys, Kayaks, telescope etc.

Epi-DIC scopes are designed for reflective surfaces e.g. silicon wafers, micro-chips, electronics etc and I have not had a chance to try one - hence no pictures, but they are much cheaper for sure. I have a used Olympus metallurgical microscope which uses epi-scopic illuminaton, but it can not be used for aquatic organisms or cells so I rarely use it. Maybe one day I will try epi-DIC.

My Zeiss DIC was straight forward to set up and no more difficult than Phase contrast. Hoffman modulation I have was finicky to set up and I much prerfer DIC - which is my favorite technique. Getting an older Fluorescence scope took some work, first finding the correct bulb was a challenge. The mercury bulbs aren't standard and those form AMscopes that I thought would work did not and I finally found a bulb from https://www.bulbamerica.com/ that worked for $200 - then I had to purchase new objectives with high NA to get brighter images.

Thank you kindly for your comments.
RB

Scarodactyl
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Re: DIC versus other forms of Microscopy

Post by Scarodactyl »

This is just a single image but I was pretty impressed with it http://www.photomacrography.net/forum/v ... 33#p269633 (the onion skin is good too but looks more obliquey to me)
Hard to say until you have a wider range of samples photographed, but I'm definitely ordering some polycarbonate to eventually try it myself. Just need to clear out some current projects first...

Robert Berdan
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Re: DIC versus other forms of Microscopy

Post by Robert Berdan »

The Sanderson picture of onion skin is spectacular and I remember reading the paper now - can't remember why I didn't try it. The paper itself does not show good pictures, but the onion skin photos by Saul was amazing for sure.
RB

Sumguy01
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Re: DIC versus other forms of Microscopy

Post by Sumguy01 »

:D Thanks for sharing the article and the photos.

Macro_Cosmos
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Re: DIC versus other forms of Microscopy

Post by Macro_Cosmos »

I personally think COL+POL produces very good results. DIC is nice, Phase Contrast is a bit specific, but both are very expensive in my opinion. Especially if you want apochromatic phase contrast objectives, they are very expensive and insanely rare.

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