If you have interest in trying to build a similar focus stacking system, I described it in detail in the amazon review for the Mitutoyo 5X microscope objective:
I was planning to do one big post of everything I learned since taking this on, but it is pretty clear than aphids deserve their own post.
Aphids are a tough group to identify. In many cases my 5X magnification is not enough, and in extreme cases I was told a 200X phase contrast microscope is absolutely necessary. Still, there are many species which can be identified from a good photo and focus stacking aphids on a slide increases that number substantially.
The first step is collecting some adult aphids, and putting them on a slide. For this I use some combination of the technique described in the USDA video on slide mounting, and the instructions which came with a slide mounting kit I bought to mount the aphids in Euparal
- Put the aphids in a ~10% sodium hydroxide solution for 12-24 hours. I am not real scientific here, and just dump some crystals in until it looks like about 10%. Unfortunately this ruined a couple when I got the concentration a bit high but I am getting a good feel for it. For a supply of sodium hydroxide, I found that some drain cleaner works.
- Squish the aphids as shown in the USDA video.
- Put the aphids in specimen clearing solution, and heat them to 120-150 degrees F for a few hours until they look clear.
- Run them a few minutes each in 70% alcohol, 95% alcohol, and 99% isopropyl alcohol.
- Put a drop of Euparal on a slide and place 3-5 aphids on the slide bottom up. After some usually futile attempts to position such that all limbs are visible, put on a cover slip.
After drying the slides I use my microscope setup to photograph with as much magnification as my setup can do.
Alata:
- Full body
- Third antenna segment
- Full body
- Third antenna segment
- Head, dorsal and ventral focus
- Apex of rostrum
- Hind Tarsus
- Abdomen, including cornicles and cauda, dorsal focus
- Cornicle, especially apical 1/4 or so.
- Cauda, especially setae number and placement
Some examples of aphids rarely reported on iNaturalist I have been able to identify since taking this on:
Metopolophium dirhodum
Myzus persicae
Macrosiphum euphorbiae
Hysteroneura setariae
Neotoxoptera formosana
Aphis craccivora
Wahlgreniella nervata
Sitobion fragariae
Eulachnus rileyi
A couple of those are likely to get pushed back to genus when someone points out a look-alike, but most of them are correct.
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