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"Factory" insect releasing

 
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Grahame



Joined: 30 Sep 2014
Posts: 50
Location: New Zealand

PostPosted: Sat Nov 11, 2017 6:27 pm    Post subject: "Factory" insect releasing Reply with quote

Hi folks,
I'm just starting work on insects that have been caught in 70% Ethanol.
This is my first time working with it.
The images being taken are stacks, to be used for identification purposes.
The problem I'm having is they are not particularly flexible.
I'm not wanting to pin them or take high res stunning shots like we see in the galleries here.
But to take "nice" shots of them with their legs moved from in front of id features and antennae visible is the aim.
Lateral, ventral, dorsal and wing detail if appropriate.
So I'm looking for a quick way of getting some form of pliability into them.
I have large amounts to deal with and want to get them through the system reasonably quickly.
2 plus days in an acetic acid etc environment is not really and option.
There are space issues as well.
Once I've posted this I'm going to try and put together some form of steam environment, it seams like the most likely thing from what I've read to now.
I'm working with arachnids, diptera, coleoptera, crane flys, acari etc so of course a few different techniques will come into play.
Have thought of Barbers (minus benzine), but Ethyl acetate is had to come by and expensive where I live.
Any thoughts folks ??
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Ichthyophthirius



Joined: 07 Mar 2013
Posts: 729

PostPosted: Mon Nov 13, 2017 10:12 am    Post subject: Reply with quote

Hi,

Can you tell us more about from what angle you approach this? There are different answers to your questions depending on your prior knowledge.

a) You do this professionally.

b) You have been hired to do this work.

c) You're a beginner, working on some kind of ecology or entomology project.

Good specimen preparation is essential in entomology. Each individual must be properly prepared (techniques differ from group to group), otherwise identification and documentation (like photography) are not possible or unnecessarily difficult.

Any shortcuts taken during sample preparation will result in time wasted during identification and documentation or even loss of specimens because pieces break off etc.

For many groups pinning is the preferred method of preparation precisely because this allows the specimen to be viewed (and photographed) conveniently from all angles.

Material stored in ethanol will be stiff; even worse when it was originally killed in ethanol as well. If any of the relaxation methods work on your sample, you're lucky.

Test a number of methods and see what works. Going through a large sample with hundreds of individuals will take you weeks anyway; there isn't really a problem with waiting 2 days for a batch to wait in acetic acid while you work on another batch.

Popular relaxation methods are dilute acetic acid (liquid or moist chamber); steam; swelling solution (for beetles I use Scheerpeltz solution (70 % ethanol, 29 % water, 1 % acetic acid; don't know what this would do to Diptera).

If you work with certain chemicals (formaldehyde, acids), the samples will no longer be suitable for DNA extraction afterwards (something to keep in mind).

One way of cutting down on the time required for this process is to exclude duplicates. If it is possible to determine the insects in the alcohol collection reliably, return all the duplicates back into alcohol and process just one or two individuals per species for photography.

Regards, Ichty


Last edited by Ichthyophthirius on Mon Nov 13, 2017 11:57 am; edited 2 times in total
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Grahame



Joined: 30 Sep 2014
Posts: 50
Location: New Zealand

PostPosted: Mon Nov 13, 2017 11:24 am    Post subject: Reply with quote

Hi Ichty,
Thanks for the reply.
While no where near a beginner, I'm not a pro either ie paid.
And no money for assistants or anybody with the skill set to call on.
This is an invert survey using malaise traps for what is primarily going to be for inventory purposes.
We are repeating it for one week per mth for the 5 mths over the summer.
3 sites are stationary and one trap will be use at a variety of sites.
We are using sites that were sampled several years ago and have another report from many yrs ago that gives us a species list.
The area is a large piece of urban wetland.
I'm used to working with much lower numbers of inverts and taking images for id etc, and have worked on all sorts of different surveys (still am for that matter)
Have been playing with macro since the early 80's and have been stacking for several yrs.
The imaging work flow is a well practiced efficient one.
The sorting is not too much of a problem, theres 2 of us with a pretty good knowledge of inverts.
Have good back up from specialists for bringing what we don't know down to sp. if possible.
Yes there will be little imaging of duplicates, less as time goes on as the sp. will already have been imaged.
The first round is the major problem as even though I have previous images of a % of them, I'm starting from scratch for this project.
Hence looking for a "quick" way to release them.
Yes they are being caught and stored in 70% ethanol.
There will never be DNA work done on them.
My "lab" studio is at home, I live in a small space with no more room to expand into.
Formaldehyde will never be finding it's way in here Smile
Again a reason to look for "quick" ways to release.
Steam looks promising and may have to resort to acetic acid.
Thanks for the Scheerpeltz solution recipe.
I guess as always invent and learn as I go when using new to me chems.
Thanks again
Grahame
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Ichthyophthirius



Joined: 07 Mar 2013
Posts: 729

PostPosted: Mon Nov 13, 2017 11:55 am    Post subject: Reply with quote

Hi Grahame,

Thanks for the update. Now I understand this much better; I shouldn't have been so sarcastic Embarassed

But yes, relaxing insects after capture in these traps is always a struggle. Beetles I would transfer into Scheerpeltz as soon as possible after capture (can stay in Scheerpeltz for years) for carding or pinning later.

Diptera I would also pin; it makes it much easier for photography and external experts would probably prefer it that way as well.

Mites usually require preparation as slides.

Arachnids I've mostly seen preserved in alcohol. It might be easiest to build a cuvette/little glass chamber and photograph it in liquid, for example alcohol or glycerol.
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Grahame



Joined: 30 Sep 2014
Posts: 50
Location: New Zealand

PostPosted: Mon Nov 13, 2017 12:14 pm    Post subject: Reply with quote

Cheers Ichty,
Will have to give the Scheerpeltz a try.
There wasn't a lot of coleoptera in the trap I've sorted and I know all bar 2.
Strangely enough I invented a curved chamber on Sun.
Cut a plastic specimen tube in half and then in half longways.
Worked well.
Have a good one
Grahame
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zzffnn



Joined: 22 May 2014
Posts: 1487
Location: Texas USA

PostPosted: Mon Nov 13, 2017 2:07 pm    Post subject: Reply with quote

Grahame,

I would suggest what Ichthyophthirius has proposed. I am not sure steaming would work more conveniently/faster than soaking in vinegar (I would suggest vinegar soak first). Maybe steam or carefully/quickly boil in vinegar, if you have to?

It would be hard anyway, since your insects were killed in alcohol. Pining them would take a long time too (sometimes, with a few insects, you can carefully bend legs and pin them; but apparently you have too many subjects to work with).
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Grahame



Joined: 30 Sep 2014
Posts: 50
Location: New Zealand

PostPosted: Mon Nov 13, 2017 3:13 pm    Post subject: Reply with quote

Hi zzffnn,
Thanks for the reply.
Boiling in vinegar ?
Not something I have thought about.
Any idea as to strength, 5% acetic ? ie pretty much normal white vinegar.
I had been thinking about adding it to the water when steaming.
Yes too many sp. to pin this first time round, I'm a one man band after sorting.
I'm just trying to get good but not perfect images.
Can work on the special images of the special finds at a later date.
Cheers
Grahame
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zzffnn



Joined: 22 May 2014
Posts: 1487
Location: Texas USA

PostPosted: Tue Nov 14, 2017 9:31 pm    Post subject: Reply with quote

Sorry, I don't know for sure, Grahame. Boiling may be too harsh for some delicate subjects. Steaming in vinegar should be more gentle.

Yes, white vinegar is usually 5%. That concentration works for fixing insects (by deactivating digesting enzymes).
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Grahame



Joined: 30 Sep 2014
Posts: 50
Location: New Zealand

PostPosted: Tue Nov 14, 2017 9:37 pm    Post subject: Reply with quote

Thanks zzffnn,
Will have a play with it, see what I can come up with.
For anyone who finds this topic sometime looking for ideas.
I don't recommend steaming crane flys (Tipulidae).
Changes their colour and doesn't loosen leg joints.
Lesson learnt Smile
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