(solved) What fluorescent stain was used on that mite?

[WalterD]

Edit: "Experiment continued-fluorescent stained Amaryllis pollen" was the previous title of this post. Here I would like to discuss different types of fluorescent stain.

As I replied in my previous post, I continued with testing fluorescent stain or to be more precise : with pollen of an Amaryllis. When adding a droplet of water with dissolved rhodamine, the nearby pollen actally jump from the stamen into the stain.
After drying they were observed with similar filter combo as the radiolaria, only this time with maximum magnification, 95x. This is a bit more difficult given the short working distance and the required immersion oil. Due to the stain the pollen were stuck to the slide, which made life easier.

As you saw at the end of my previous post, the stain is initiating red emission, with increased brightness, contrast and detail according my observation. The tiny -65 mu- pollen are completely absorbed by the drop of stain. Result was an interesting observation. The distinct ridges on the surface I was familiar with, however they showed a bead-like structure I have not seen before. Just checked it in an online SEM picture https://www.sciencesource.com/archive/A ... 30383.html and did not notice it there either. The colours in the final image were tweaked a bit with Photoshop, stacking was done with Zerene (dmap).

[Jacek]

SUPER !

[WalterD]

Glad you like it Jacek, you're my biggest fan
I continued with another specimen (a mite) and a different stain.
The riddle is : what kind of stain did I use on this mite. Hint: it rhymes with mite.

Don't forget to visit my homepage for double resolution.
www.waltermachielsen.com

PS: one of the aft legs was out of focus and not included in the picture.

[ChrisR]

Ethidium Bromide ?

[Pau]

The distinct ridges on the surface I was familiar with, however they showed a bead-like structure I have not seen before. Just checked it in an online SEM picture and did not notice it there either.

Nice images and interesting observation. The similar but bigger Lilly pollen grains shows it both with fluorescence and SEM
http://www.photomacrography.net/forum/v ... 041#215041

Please, see PM

[Jacek]

[WalterD]

Glad you like it. I used Calcofluor White, not part of the original kit. What was in the kit was "optical brightener" (25ml), just discovered when cleaning up my cupboard. I read it works more or less the same as calcofluor, have to try it. Calcofluor White is used for staining cellulose and chitin. Already had interesting results on botanical sections, better to use that for next post.

So I wanted to continue with staining mites and show you "before" and "after" pictures, to illustrate the effect. What actually happened was that the mites I selected turned out to be autofluorescent, so no further staining was required. Both images of dust mites @ 17x (respectively ventral and dorsal view) show fluorescence combined with understage light, to present details of the isolated hairs (that are like sensors for the mite)

The similar but bigger Lilly pollen grains shows it both with fluorescence and SEM

Yes, I remember that, nice set of fluorescence pictures, in particular the 40x/0.65 one. I should give it a try with my fresh lillies I guess.

[carlos.uruguay]

Nice!!!!!!!!!!!!!!!!!!!!!!!

[WalterD]

Thank you Carlos!

I've added 3 50% crops, as some details disappear when resizing to 1024 pixels (compared in 3rd image) . Have a look at the shape of the hairs (possibly with small scales) the tools on the mouth parts and the structure of the antenna shaped legs.

[carlos.uruguay]