Genius Well done that manOriginally Posted by Ruary
it's £12
Ruary
you had a post on BBKA a couple years ago on making stained permanent (ish) nosema slides
what is your latest advice on making them ? or is it not worth bothering with?
Genius Well done that man
it's £12
Ruary
you had a post on BBKA a couple years ago on making stained permanent (ish) nosema slides
what is your latest advice on making them ? or is it not worth bothering with?
Last edited by The Drone Ranger; 11-04-2013 at 06:01 PM.
Well I suppose I could say come to Gormanston and ask me there, but I am feeling kind so my process is to place a drop of distilled water containing the spores on the sldie and add nigrosin stain. Spread the drop so that it covers the extent which would be covered by the cover-glass. Dry it on a hot plate until almost dry and then mount in plain glycerine jelly. Check that it is OK. allow to set etc and seal with whatever varnish you use.
I am wondering whether it would work with drying the slide completely.
Last edited by Ruary; 11-04-2013 at 06:03 PM. Reason: typo
I'm No expert Ruary
This method dries the specimen completely but they are fixed in ethanol so that would seem to be an important step and I don't think the slides would store
When air-dried, ethanol-fixed smears of infected tissue are stained with Giemsa’s stain (10% in 0.02 M phosphate buffer) for 45 minutes.
Nosema apis spores will have a distinctive appearance, with thick unstained walls and an indistinct blue interior, without visible nuclei. Insect cells, fungal spores and other
protozoa stained in this way will generally have thinner walls, blue/purple cytoplasm and magenta-coloured nuclei
Clipped from http://www.oie.int/fileadmin/Home/en..._NOSEMOSIS.pdf
No wish to spoil your fun, but wet (water) mounts of bee abdomen juice are fast and easy. When Nosema is there it is usually abundant and really obvious - those jiggling rice grains. OK, you can't keep the slides, but that's all I've ever done. Pollen is different. You can usually see the structure better if it is stained in one way or another.
It might be interesting to make dry mounts of Nosema to see if you can spot the difference between the fat ones (N. ceranae) and the thinner ones (N. apis). I think.
Yes, fresh is best but what if you need to show nosema and don't have a sample of infected bees. That is what my method is designed to do.
DR I have a home lab no Giemsa but plenty of nigrosin, I tried the glycerine jelly as an experiment and it has worked. It won't cater for examination at X1000 but the thickness of the mount is OK for X400.
Gavin I think you might have your nosemas reversed apis is more square ended, nosema ceranae is slighly almond shaped. See next post
Last edited by Ruary; 12-04-2013 at 08:01 AM.
nosema combined.jpg This shows the two nosemas at the same magnification the sample of nosema ceranae obtained via Dave Cushman from FERA, the Nosema apis from my collection and PCR'ed by Randy Oliver in USA who was looking for a sample of genuine Nosema apis.
Last edited by Ruary; 12-04-2013 at 08:02 AM. Reason: added material
I did indeed - thanks for putting me right.
Here's an interesting aside. N vespula is of course a wasp parasite.
My first attempt at looking for Acarine.
Acarine Dissection.jpg
Nice dissection, you removed the collar so that the whole of the tracheae can be examined. Now the real question positive or not??
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