|From:||"kevin williams" |
DAKO makes an aqueous mounting medium called Faramount. It works great.
It's liquid at RT but it dries hard when coverslipped.
We love it.
Hazel Horn, HT/HTL (ASCP)
Arkansas Children's Hospital
When using AEC as a chromogen, I had good results using Crystal/Mount which polymerizes and can be viewed without a coverslip or can be post-mounted with regular mounting medium and coverslipped.
Now I specialize in muscle biopsies and some enzyme stains require aqueous mounting. I got plenty of bubbles with Aquamount and glycerol/gel. I switched to Crystal/Mount and let it polymerize at room temp and then coverslip and the bubble problem was eliminated.
Mary North, HT(ASCP), HTL
Oregon Health and Science University
I think that all aqueous mountants are a pain, but I find that Dako's Glycergel is user friendly. I put into warm water and let it melt completely, then invert the bottle with the cap on to let the bubbles rise, then take off the cap and dispense just enough onto the coverslip. If you still get air bubbles you can place the slide onto the slide drying hot plate for just long enough to soften the mountant and the bubbles run off without much trouble.
Can't help regarding your kit though. My experience with fast red is with the Sigmafast tablets which seem to go off if left for too long, I use them within 10 minutes of preparation.
University of Cambridge
several things: 1. aqueous mount try to add some disH2O, use probe to pop bubbles, put glue on the coverslip not the slide then drop DisH20 onto the glue, touch slide to the coverslip ...try it it works great.
2. The argument for overnight incubation is to save $$$ on primary antibodies by diluting them way out, otherwise you can find procedures that can be done start to finish in 1-2hrs with low cost and great consistency.3. Melanoma cases will vary significantly as far as the positivity of HMB45 for each lesion; the S100 should be more consistent. We run HMB45 with and without AR this may prove interesting to review.
4. Fresh solutions are great but you want to be able to use solutions that you can make up in advance and put into a refrigerator to store and reuse later. So, I am pro DAB which doesn't fade, however it is brown and not preferred for the dark Derm lesions... there is a " so called permanent mount by Novacastra for RED"
5. The antibody companies : Dako, Cell Marquee, Biogenex, Ventana, Zymed, Novacastra, Biocare, Signet can all do better than Thermo in my book... review as much as you can and balance the $$$ with the reproducible quality. I did use the Shandon system by hand many moons ago and it was OK for very small numbers of antibodies, but the other guys can just beat them out for the " best results " every time.
Just my two bits MATE,
Be Well young man, contact me anytime... RichRich Your email address dose not work. KEVIN