Mounting *very* small beetles for macro stacking

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Beatsy
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Mounting *very* small beetles for macro stacking

Post by Beatsy »

I managed to pick three tiny beetles out of some sphagnum moss today and they are currently wandering around in a small sample bottle. My problem is that they are barely 0.3-0.4mm long and I'm not sure how to proceed from here. They are very interesting to look at under magnification (like glossy, jet-black ladybirds with slots in the lower edge of the carapace where legs and antennae poke through) and I want to stack them in a near-lifelike pose, preferably on the sphagnum moss leaves they were picked from. The 50x Mitty will get an airing for these as they really are that small.

But how to handle and mount them? Freezing might work as a first step but if the legs curl I can't fathom how to straighten them at this scale. Similarly, I thought of using ethanol instead, but I suspect it will have the same effect as freezing once the alcohol evaporates. Then there's the physical handling to get them mounted - I may be "re-purposing" my diatom arranging gear to achieve this.

So, I'm going to leave them wandering about 'in-vitro' for tonight and have a go tomorrow. In the meantime, am I taking this beyond the realms of practicality, or has anyone else managed to mount and stack insects at this scale? Hints, tips or comments gratefully received and please feel free to post pics on this thread too if you have examples to show. Thanks.

rjlittlefield
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Post by rjlittlefield »

Since they are currently still alive, and you have several, I suggest testing one to see how it reacts to progressively colder environments. I would not be surprised if cooled gradually to near freezing, it retains a natural pose but stops moving. Rather like those dew-covered flies that we see photographed by John Hallmén and others.

--Rik

zzffnn
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Post by zzffnn »

The beetle book I have says that killing by ethyl acetate will keep beetles relaxed for several days or weeks. The same book also says that you can relax dry beetles by quickly soaking in warm water + a drop of dish soap, or putting them in between tissue paper saturated with warm water + moth ball.

I remember small (5mm) lady beetles freshly drown in white vinegar have relatively flexible joints. You can drown them directly in white vinegar, which itself is used in entomology as a (color) fixing agent. An entomology professor told me to use that for preserving caterpillars (which can be put in live and stay preserved for months that way). Though beetles' colors are mostly structural, so they won't have much to fade to start with.

You can also kill them quickly by generous amount of acetone. That may take less than an hour. Small (5mm) lady beetles killed this way do curl up their legs with slight rigidity, but you can still spread their legs if you patiently do so under a dissecting scope. But with a 0.5mm beetle this may be difficult. I would think drowning in white vinegar would work better.

Whatever you end up doing, please report back with your findings :twisted:
Last edited by zzffnn on Thu Nov 23, 2017 10:35 am, edited 1 time in total.
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johan
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Post by johan »

It's not really the killing that's the challenge, it's the position afterwards. I share your pain - manipulating these through relaxation isn't a task for the easily frustrated :)
My extreme-macro.co.uk site, a learning site. Your comments and input there would be gratefully appreciated.

Ichthyophthirius
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Post by Ichthyophthirius »

Hi,

0.4 mm is very small; Ptiliidae fall into that range (Nephanes titan is the smallest UK beetle at 0.55-0.65 mm).

Ptiliidae are usually mounted in resin on slides. With a lot of patience and skill, they can be carded which looks like this: http://www.markgtelfer.co.uk/beetles/ptiliidae/ You use very fine paint brushes and insect needles to arrange them like this.

If you want them to look life-like, the best way is to keep them alive and slow down their movements in the cold and/or CO2 (baking powder + citric acid).

When you say "glossy, jet-black" I immediately think of moss mites: http://www.alexanderwild.com/Insects/In ... /i-rxchfdB

Regards, Ichty

Lou Jost
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Post by Lou Jost »

What if you were to put them in glycerine + water + surfactant and then freeze them? They would slowly die but maybe the glycerine would keep the legs from curling, if the solution were thick enough. Then perhaps the glycerine could be slowly replaced by ethanol (by adding ethanol to the solution and removing glycerine) and left to evaporate, leaving your specimen....

Beatsy
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Post by Beatsy »

Lou Jost wrote:What if you were to put them in glycerine + water + surfactant and then freeze them? They would slowly die but maybe the glycerine would keep the legs from curling, if the solution were thick enough. Then perhaps the glycerine could be slowly replaced by ethanol (by adding ethanol to the solution and removing glycerine) and left to evaporate, leaving your specimen....
This sounds quite feasible Lou. I wish you'd posted this about 10 hours earlier. Before I opted for the simple freezer route :(

No big deal really. Tomorrow will reveal if straight freezing was a mistake and I know where to get more if it was.

Thanks everyone else for the comments too. These may be mites rather than beetles after all, but they don't look like the common moss mites - the slots in the carapace are definitely different. Unless they're just mechanical damage. Again - tomorrow will reveal all - or nothing :)

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Post by Lou Jost »

I hope one or more of these methods work for you!

Beatsy
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Post by Beatsy »

Update (while the glue dries): I'm pleased to report that these mites/beetles are so rotund and their legs so short, they came out of the freezer looking pretty good. The legs aren't long enough to curl round the body and tangle, and the antennae stayed pretty much straight and natural-looking too.

On the down side, I already lost one by breathing too heavily :( Luckily, the remaining two were the best examples anyway.

On the premise that there are more of them in the sphagnum sample, I decided to skip mounting on a moss leaf until the next attempt. Too much risk of loss or damage - and I'd have to do a much deeper stack to get the moss in focus (not much fun at 50x and 1 micron steps). So for this first attempt I've simply glued them to the points of 00 ent pins using a microscopic dot of PVA. The pins come in at an angle from the side and should be pretty easy to hide or edit out in the final image. Stacking next - I'll post pics in the studio macro section when it's done.

Beatsy
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Post by Beatsy »

Hmm, struggling to light these nicely. I managed to ping one off the pin and straight up my left nostril too (PVA doesn't hold quite as well as it appeared to :) ). The one remaining specimen is now on the back burner until I figure out a better way of lighting it. Couple of test stacks here to conclude (for now). First is 20x Mitty, APS-C mode, 300 images. Second is 50x, also APS-C, 650 image stack. Nothing special, but enough to ID them (probably common moss mites). This one is just under 0.5mm long.
Image
Image

tevans9129
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Post by tevans9129 »

Fascinating !! What lighting are you using for these shots?

Beatsy
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Post by Beatsy »

tevans9129 wrote:Fascinating !! what lighting are you using for these shots?
Thanks. This used a "double diffused ping pong ball". Just a ping pong ball with two holes in (one for the objective to look through and the second to introduce the specimen from underneath). This is surrounded by a second, larger dome diffuser with various halogen sources lighting that. I think I need a smaller ping-pong ball (or equivalent) to get the exact lighting I'm after. Struggling to apply much light from the front with the shorter working distance of 20x and 50x mitties. Something around 15mm diameter should help that too. That blimmin' inverse square law plays havoc with blown highlights when you're using such small subjects. I'll have another go once I find a suitably-sized diffuser.

tevans9129
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Post by tevans9129 »

Thanks for the info Beatsy...being a rank rookie, I would never have guessed ping pong balls. Do you have any images of your setup for thses shots?

Beatsy
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Post by Beatsy »

tevans9129 wrote:Thanks for the info Beatsy...being a rank rookie, I would never have guessed ping pong balls. Do you have any images of your setup for thses shots?
This is the latest shot of my rig, though with only the ping pong ball in place. The secondary dome diffuser is at the back on the right - it's just placed to surround the ping-pong ball. If you plan to try ping pong balls yourself, get top quality ones. Cheaper ones are thicker walled (variably so) and let much less light through.
Image

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Post by Saul »

Both stacks are IMPRESSIVE :shock:
Saul
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