Very interesting photo of a testate amoeba

[Pat Thielen]

This is a very interesting, and different, sort of photo (in my opinion anyway). This is a testate amoeba taken using an LED light lit from above. It is also a stacked image, which gives it some depth of field. I thought I'd try lighting things from above using an LED light that I bought from Ikea (thanks to a suggestion from a forum member). It actually worked better than I had thought; I like how the test has a very glass-like texture and how it just kind of floats there in space.

The sample of pond scum I took from that small lake is proving to be a very good source for testate amoebas. I think I've seen three different species so far.

As always, let me know what you think and if you have any suggestions for improvements.

[JimT]

I agree. Worked out nicely.

[billbillt]

The surface quality of your subject is very interesting.. Great job!...

BillT

[mrsonchus]

Very intriguing image Pat, it really does seem to float-in-space, even looks a little spooky! Very nice and very interesting - looking at this image I find myself staring closely to glean extra almost hidden details - unusual and crystal-clear too!

[zzffnn]

Thank you for sharing, Pat.

If your amoeba test was imaged in water, you may try to remove all water and image it again. That will provide even better contrast, as the refractive index difference is more between test-air than test-water.

Is your 20x NA 0.45 objective a long working distance one? 0.17mm corrected or no cover type? No cover type objective is preferred, if you image the test dry in air. I also wonder if you had to use long exposure to obtain enough light, as a non LWD NA 0.45 objective may block most overhead light.

[Pat Thielen]

I agree. Worked out nicely.

Thanks!

[Pat Thielen]

Very intriguing image Pat, it really does seem to float-in-space, even looks a little spooky! Very nice and very interesting - looking at this image I find myself staring closely to glean extra almost hidden details - unusual and crystal-clear too!

Thanks John. I find myself doing the same thing -- Seeing if there are any little details I can see. I think it's also cool that you can look right into the test as well; it's like looking into an ornate glass pitcher. One thing I'm considering is adjusting the color balance a bit as it is obviously shifted. At first I liked it, but I'm thinking I may like it better with more natural looking color. Thanks for looking!

[Pat Thielen]

Thank you for sharing, Pat.

If your amoeba test was imaged in water, you may try to remove all water and image it again. That will provide even better contrast, as the refractive index difference is more between test-air than test-water.

Is your 20x NA 0.45 objective a long working distance one? 0.17mm corrected or no cover type? No cover type objective is preferred, if you image the test dry in air. I also wonder if you had to use long exposure to obtain enough light, as a non LWD NA 0.45 objective may block most overhead light.

I hadn't thought about removing all the water; that may be something to try! All my objective are infinity / .17, so they are calibrated with a cover glass in mind. I'm not sure if my 4x / .1 lens is or not, but it isn't one I use for most of my photography anyway -- It's more of a "hunting" lens. The working distance of my lenses are really quite close so it is a challenge getting the light in there. You're right -- The lens itself does block a lot of the light. I had to move it around to find the best position for it. I was actually able to use this technique, to some degree, with my 40x / 0.65 lens. This surprised me a bit as I didn't there would be any way for me to sneak any light in there at all. I'll have to do more experimentation all around, but I am liking how it looks so far. I may look for a much smaller light source on a flexible neck; I've heard tell of such things. But I am happy I got what I did; I think it's off to a good start.

Next thing to do is to start combining traditional under-stage lighting with above stage lighting to see how that work. I did use this with darkfield which worked not too badly.

Can anyone recommend a bright, small, flexible light that could be used for over-stage lighting? It would be really cool if I could find one that could run parallel to the lens and shine light more or less straight down onto the slide. That would help eliminate flare from the cover glass.

Thanks for your suggestions; I do appreciate it!

[Pat Thielen]

Thanks to Josh's response to my other testate amoeba post, and his website, I believe this is a species of Mediolus. And, he was actually home although you can't see any of the psuedopods in the photo. Also, this guy was bigger than the other testate amoebas I came across in my exploration (actually, about twice the size of the others I observed). I really like the jagged aperture; it makes him look even more fierce than he already is!

[charlie g]

Your lighting here evokes the dark power of Goya paintings, Pat. This could be the crown renched from a cruel despot and tossed to the ground...I complete your picture to right of this screen! charlie guevara.

It's been years..and I still have not utilized my nightvision monocular ("Night Hawk" brand device) with the microscope to see if protists and meiofauna behave differently in low light situations. I always wonder of how unnatural the powerful lightblast we bathe our hydras, water fleas, rotifers, and others in..how it may strongly alter their behaviors and body forms.

LED small lights are very useful for my microscopy, thanks again, Pat.

[zzffnn]

Pat,

Sorry to be so blunt - you were using your 0.17mm corrected objective out of spec (without cover slip). Though your image looks fine.

Using light parallel to objective and straight down onto slide will actually produce unwanted flare. Nikon made BD series no cover objectives, which allow through the objective lighting: http://www.optotek.net/Microscope_Inven ... tives.html

But they are prone to flare and need polarizers at the very least. Instead, most macro/micro photographers use long working distance no cover objectives and oblique incident light.

I would suggest using some zoomable LED torch at high angle (close to perpendicular with objective) and wrap some paper cone around objective as diffusion. Shine LED onto paper cone.

I have tried 4 Maglite Solitaire LEDs. They can rest on your scope stage, with help of some mounting putty.

This more powerful torch is bigger and heavier, so it
needs a gooseneck holder:

https://www.amazon.com/gp/product/B00SW ... UTF8&psc=1

https://www.amazon.com/gp/product/B00G3 ... UTF8&psc=1

You may need at least two sets of light and holder for even lighting, preferably 3 sets.

[JimT]

Can anyone recommend a bright, small, flexible light that could be used for over-stage lighting? It would be really cool if I could find one that could run parallel to the lens and shine light more or less straight down onto the slide. That would help eliminate flare from the cover glass.

Some body on this forum has used a half ping pong ball with a hole cut in the top for the objective lens to reflect bottom light and also illuminate the top. As I remember the image was very good but my sense is it is mostly for lower magnifications.

I haven't tried it yet but the next time I am at my Son's house I am going to take one of his ping pong balls.

JimT

[Pat Thielen]

Pat,

Sorry to be so blunt - you were using your 0.17mm corrected objective out of spec (without cover slip). Though your image looks fine.

Using light parallel to objective and straight down onto slide will actually produce unwanted flare. Nikon made BD series no cover objectives, which allow through the objective lighting: http://www.optotek.net/Microscope_Inven ... tives.html

But they are prone to flare and need polarizers at the very least. Instead, most macro/micro photographers use long working distance no cover objectives and oblique incident light.

I would suggest using some zoomable LED torch at high angle (close to perpendicular with objective) and wrap some paper cone around objective as diffusion. Shine LED onto paper cone.

I have tried 4 Maglite Solitaire LEDs. They can rest on your scope stage, with help of some mounting putty.

This more powerful torch is bigger and heavier, so it
needs a gooseneck holder:

https://www.amazon.com/gp/product/B00SW ... UTF8&psc=1

https://www.amazon.com/gp/product/B00G3 ... UTF8&psc=1

You may need at least two sets of light and holder for even lighting, preferably 3 sets.

Thanks for your response. Actually, I was using a coverglass when I took the image as I don't have a water immersion lens. So, that part was in spec with the lens. I did have to move the light around a bit to find the best angle -- I could watch through the scope as I did this so it wasn't too difficult. I would like to try brighter lights; I thought of the Maglite idea some time ago but I haven't put any to the test yet (no pun intended but what an awesome geeky pun!). That may be my next experiment; using multiple Maglites and lighting the stage as I would (more or less) a studio portrait. That'll be fun -- Hopefully I'll be able to get to that soon and post my results.

Thanks again for your response; I really appreciate it!

[Pat Thielen]

Can anyone recommend a bright, small, flexible light that could be used for over-stage lighting? It would be really cool if I could find one that could run parallel to the lens and shine light more or less straight down onto the slide. That would help eliminate flare from the cover glass.

Some body on this forum has used a half ping pong ball with a hole cut in the top for the objective lens to reflect bottom light and also illuminate the top. As I remember the image was very good but my sense is it is mostly for lower magnifications.

I haven't tried it yet but the next time I am at my Son's house I am going to take one of his ping pong balls.

JimT

I remember that thread but I can't recall the details or who posted it. And I think you're right -- It was intended more for lower magnification. Another thing to try!

[Pat Thielen]

Your lighting here evokes the dark power of Goya paintings, Pat. This could be the crown renched from a cruel despot and tossed to the ground...I complete your picture to right of this screen! charlie guevara.

It's been years..and I still have not utilized my nightvision monocular ("Night Hawk" brand device) with the microscope to see if protists and meiofauna behave differently in low light situations. I always wonder of how unnatural the powerful lightblast we bathe our hydras, water fleas, rotifers, and others in..how it may strongly alter their behaviors and body forms.

LED small lights are very useful for my microscopy, thanks again, Pat.

Thanks for your comments. Yeah, it does have a painterly feel to it. I hope I can make more like this.

I would be very interested to see what you discover regarding low light behavior will be. I'm sure they aren't used to ridiculously bright light shining up at them. Not all have photo-receptors but certainly some animals do. When you have a chance to experiment with this let us know!

[gekko]

Wonderful that you are experimenting and getting such beautiful results.

[Pat Thielen]

Wonderful that you are experimenting and getting such beautiful results.

Thanks!