another look at BMSB highly sculptured cuticle

Images taken in a controlled environment or with a posed subject. All subject types.

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rjlittlefield
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another look at BMSB highly sculptured cuticle

Post by rjlittlefield »

A few weeks ago I showed you a first look at "Highly sculptured cuticle around stink gland of BMSB" (Brown Marmorated Stink Bug). The most informative image that I could get then was using a 100X NA 0.80 objective that produced so much false color that eventually I showed only its green channel as B/W.

I recently received a Nikon 40X NA 0.80 M Plan apochromat, which I hoped would give a much better image. It required some surgery, which I have described separately, but indeed the image is much better.

So, here we have the same specimen in color.

Let's start with the highest mag stereo pair. This is actual-pixels crop from a Canon T1i, heavily sharpened, using Photoshop USM first with 85% at 2.3 pixels, then 200 pct at 1 pixel. Field size here is about 0.12 mm high by 0.06 mm wide. Stereo separation is 12 degrees, which for me makes the structure more clear, at the cost of exaggerating the depth relief.

Image

Here is the whole frame area from which the stereo pair was cropped.

Image

For context (and so I can find this area again if I want to!), here is a sequence of images: 2X, 10X, 40X, and 100X equivalent crop. In each image, the rectangular outline shows the area of the next higher magnification image. The outline in the last image shows the area of the stereo pair.

Image

A keen observer will notice that the area I've chosen here for stereo pair is not quite the same as I used previously with the other objective. The reason for that choice is that I thought this crop gave a more clear impression of how the sculptured cuticle is structured, since some of the protrusions are seen almost sideways.

If you'd like to peruse the whole frame area as a stereo pair, grab the image HERE (10 MB) and run it through StereoPhoto Maker.

I hope you find this interesting!

--Rik

Edit: link to description of the objective's surgery
Last edited by rjlittlefield on Thu Jan 23, 2020 5:30 pm, edited 1 time in total.

rjlittlefield
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Post by rjlittlefield »

Shooting info: Canon T1i camera (sensor size 22.3 x 14.9 mm, 4752 x 3168 pixels), ISO 100, illuminated by 3 flashes at 1/16 power, diffused through one layer of kleenex tissue, mirror lockup, 1/200 sec shutter. 251 frames at 0.25 micron, so total field size 0.56 mm wide by 0.37 mm high by 0.062 mm deep. Stereo separation +-6 degrees = +-1.18% .

--Rik

abpho
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Post by abpho »

I can view stereograms just fine. Whether that is in print or on the monitor. But these stereo-pairs don't work out for me.
I'm in Canada! Isn't that weird?

rjlittlefield
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Post by rjlittlefield »

abpho wrote:I can view stereograms just fine. Whether that is in print or on the monitor. But these stereo-pairs don't work out for me.
Interesting! I wonder if I've chosen a separation that exceeds some limit for you.

I would be curious to know if you can handle all the pairs at https://www.photomacrography.net/forum/ ... hp?t=33871 .

--Rik

David Sykes
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Post by David Sykes »

It takes a slight effort but I can view that image just fine, including the space ship emerging from the black hole !

Incidentally, to show friends your stereo images you can obtain cell phones with lenticular screens.
Just before Xmas, Masuji SUTO (SPM author) gave me a PPTV King 7s phone, the stereo display is good.


David

MacroLab3D
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Post by MacroLab3D »

Just wanted to let you know that your 3D shots all work for me no problems and it is my favorite format when i can just cross my eyes and see immediate result. Especially impressive high magnification ones.
-Oleksandr

Troels
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Post by Troels »

Very interesting investigation.
This stereo is fascinating, but needs a little more effort from my side than usual.

I suppose two resons:
1. The immage is big. Using a big screen I haveto lean back to get a little more distance than normal.

2. There are numerous repeated features along the vertical axes. That confuses my focusing. Which one to choose? The black contours around the single pictures are of great help. I use them to focus on.
Hope this can help somebody.
Troels Holm, biologist (retired), environmentalist, amateur photographer.
Visit my Flickr albums

Ultima_Gaina
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Post by Ultima_Gaina »

I don't mean to hijack your thread with my own stuff, but, coincidently, I also did an x4 stink bug macro this weekend, and now I see your post.

Image

.. and here is a full-size view:

Image

I thought that the white spots ("balloons") you flagged in your previous post were dust particles.
I gave the bug an ultrasonic bath before the photo session. It looks to me that some of them were dislodged, but many remained trapped in those volcanos.
So aren't these shiny "balloons" dust particles, after all?

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Post by rjlittlefield »

Ultima_Gaina wrote: I gave the bug an ultrasonic bath before the photo session. It looks to me that some of them were dislodged, but many remained trapped in those volcanos.
So aren't these shiny "balloons" dust particles, after all?
It may be that you had some dust also. But most of what you're showing here do look like the "balloons in pits" that I documented in earlier threads, https://www.photomacrography.net/forum/ ... hp?t=40448 and https://www.photomacrography.net/forum/ ... hp?t=40624 . I still have not learned what their function is.

As illustrated by your image, there seems to be a smoothly graded distribution of shapes. They range from fully expanded "balloons" as shown in my earlier threads, to the very narrow post-like shapes that appear in the high mag shots of my thread here. I like David Sykes' description of "the space ship emerging from the black hole". But I suspect the projection has uniform color for its entire height, and the base is simply invisible in the strong shadow created by side lighting.

--Rik

Ultima_Gaina
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Post by Ultima_Gaina »

Rick, I still have the bug pinned in from of my camera and I used an x10 objective this time, to capture a dorsal view of the stink bug.

Image

I can see more clearly the ballons at the bottom of those pits. But these are not white.

The shiny white stuff still looks very close to dust particles to me, when zoomed in at 100%. Maybe the particles get a spherical shape while moving inside the pits and hitting the walls. I can see particles inside the pits that are not spherical, and sometimes there is more than one inside, on top of a dark "balloon"

Image


Image

Image

BugEZ
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Post by BugEZ »

I very much enjoy this thread. Stereo works fine. Shape and arrangement of whitish “corpuscles“ in Rik’s stereo I find as interesting as the balloons in previous threads.

A Christmas cactus I brought up from my basement had a hitch hiker stink bug onboard. I smiled as I eyeballed the pits on this true insect. It chose to remain silent on the “balloon in pit” topic. After showing my wife the blossoms the plant and passenger were returned to the basement ecosystem unharmed. I see some damage (puncture wounds) on some of my assorted Christmas cactus, but they continue to thrive. I have a non aggression treaty with the stink bugs.

Keith

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Post by rjlittlefield »

Ultima_Gaina wrote:Rick, I still have the bug pinned in from of my camera and I used an x10 objective this time, to capture a dorsal view of the stink bug.

<image snipped>

I can see more clearly the ballons at the bottom of those pits. But these are not white.

The shiny white stuff still looks very close to dust particles to me, when zoomed in at 100%. Maybe the particles get a spherical shape while moving inside the pits and hitting the walls. I can see particles inside the pits that are not spherical, and sometimes there is more than one inside, on top of a dark "balloon"

<images snipped>
Sorry for the delayed response; I did not see your post until just now.

Your photos are interesting and I sympathize with your doubt.

It's hard to tell for sure what's going on at this limited resolution, but I'm pretty confident that you're looking at two different sorts of "white stuff".

The two unusually large white blobs that you've circled in your image look to me like debris. The lower one could be a large balloon, with another chunk of white debris sitting in front of it.

Most of the other white things, the ones that are centered in their pits and have narrow bases, broader tops, look to me similar to the balloons that I imaged. I have marked the more definite of these on your image with yellow circles.

Image

In addition you're showing a lot of pits that look empty, nothing projecting from the bottom of them. I've marked some of those with cyan. I think that these are what you're describing as "dark balloons", but as far as I can tell there's actually nothing in those pits. The dark rounded shapes are just from the shape of the pit, either non-reflective or more likely reflecting dark parts of the environment.

My specimens have those too, but mostly in places farther back on the body. I have no idea what determines whether a pit develops an expanded balloon, a collapsed balloon, a compact post (as seen in the sculptured cuticle images), or nothing at all.

Here I have marked some empty pits on mine. There are not many.

Image
I don't mean to hijack your thread with my own stuff
The problem I see is that we're now discussing balloons in a thread about structured cuticle. I suggest that we split the discussion about balloons into a separate thread with an appropriate title, so that it can be found and referenced more easily. I can do that as Admin, after you've seen this so that you know what's going on.

--Rik

Adalbert
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Post by Adalbert »

Hi Rik,
I have already taken some photographs of the true bugs but never seen such balloons (only empty pits) :-(
Probably I cleaned, scrubbed them too intensively :-)
In the case of the cuticle I have only observed something like that up to now:
https://www.flickr.com/photos/156413690 ... ed-public/
BR, ADi

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Post by rjlittlefield »

Adalbert wrote:I have already taken some photographs of the true bugs but never seen such balloons (only empty pits) :-(
Probably I cleaned, scrubbed them too intensively :-)
The balloons on my specimens seem pretty tough. They're not vulnerable to any solvent that I tried (as described at https://www.photomacrography.net/forum/ ... 005#255005). Ultrasonic cleaner didn't touch them either, and the pits are so small that I don't have any brushes that would reach into them. I think I would have trouble getting rid of my bugs' balloons if I wanted to, though I have not specifically tested that.

So, my guess is that your bugs never had balloons to start with. There are big differences between species, sometimes even big differences between individuals of the same species. Were you looking at Brown Marmorated Stink Bug, Halyomorpha halys? (I searched the forum, and the only posts I see of yours for "bug" were showing a western conifer seed bug, which is a very different beast.)
In the case of the cuticle I have only observed something like that up to now:
https://www.flickr.com/photos/156413690 ... ed-public/
The cuticle in that image shows several types of common patterns. You might be interested to look at "A Glossary of Surface Sculpturing", which can be downloaded for free from https://zenodo.org/record/26215#.XY05sy70nhd . The images begin on page 26.

Unfortunately I do not see anything in the 44 SEM pictures of that publication that matches well with the sculptured cuticle that I have photographed here.

The closest match I know is from Thomas Eisner's book, "For Love of Insects", page 234, where this image appears:

Image

At first impression, the sculpturing shown by Eisner in his SEM image of Chelinidia vitiger (bottom right panel) does not look much like what shows in my optical image of Halyomorpha halys. But I do not know how much of that difference is a matter of geometry, versus a difference in imaging technique. I've approached the local university to see if they could help me get an SEM image of my specimen, but no reply yet on that route.

--Rik

Adalbert
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Post by Adalbert »

Hello Rik,
So, my guess is that your bugs never had balloons to start with
yes, probably.
So, I have just checked the next two pictures but haven't found any balloons :-(
https://live.staticflickr.com/4230/3541 ... aacd_o.jpg
https://live.staticflickr.com/4254/3541 ... af21_o.jpg
but sometimes small, white points in the pits.

Unfortunately I do not see anything in the 44 SEM pictures of that publication that matches well with the sculptured cuticle that I have photographed here.
maybe at the top of the picture 19?

And thank you for the interesting links!

BR, ADi

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