Nematode spicules tinted with eosin y

[lperepol1]

https://flic.kr/p/2nRaXt1

[macnmotion]

Looks like you got your nematode staining figured out? Nice photo.

[dtsh]

Nice image, just need some cheesy music and a terrible script.....

Some details on the process would be appreciated. Most of my dealings with nematodes are as eggs.

[lperepol1]

Nematode Staining Experimental.

Fix nematodes hot 90°c in TAF or straight TissuePro (https://tissueprotech.com/fixative-solu ... 32-oz.html)
Let them soak for a month in fixative.
Run fixative and nematodes through a fine 43-um nylon sieve.
Wash nematodes off with 100 ml of distilled water.
Add 4 ml of modified Harris hematoxylin to the 100 ml of distilled water and nematodes.
Should be translucent, not dark.
Soak for 60 mins.
Run hematoxylin and nematodes through a fine 43-um sieve.
Wash nematodes off with 100 ml of distilled water.
Add 1ml of eosin y to the 100 ml of distilled water and nematodes.
Soak for 10 mins
Run eosin y and nematodes through a fine 43-um sieve.
Mix 2 ml of 95% lactic acid, 2 ml glycerin, 1 ml of 95% ethanol and 100 ml of distilled water.
Use the mixture to wash nematodes off of the sieve.
Let water slowly evaporate off until nematodes are in pure glycerin. (2 weeks to a month at room temperature and a fan blowing over the container)

[lperepol1]

Histological stain (Modified Harris Hematoxylin and Eosin Y counter stain) on a broken nematode.
If the stains get under the cuticle, they work well. This could be just luck, though.
https://flic.kr/s/aHBqjAc8mi

[LouiseScot]

Seems to me you need a lab to do all those procedures!
Louise