Best mounting medium for microscopy?


#1

I’ve been using Prolong Gold and I’m not a huge fan.

What mounting media do people prefer for spinal cord or DRG?


#2

We are huge fan of the Mowiol mounting medium.
Good results at a cheap cost!

Protocol:

  1. Add 4.75 ml (6 g) of glyercol in a 50 ml centrifuge tube and add a small stir bar.
  2. Add 2.4 g of Mowiol and stir to mix.
  3. While stirring, add 6ml distilled water and leave for 2 hours or overnight at RT.
  4. Prepare 0.2 M Tris Buffer pH 8.5 ( Add 2.44 g of Trizma to 800 ml distilled H2O, adjust pH to 8.5 with HCl, adjust volume to 1000 ml with distilled H2O)
  5. Add 12ml 0.2 M of Tris (ph 8.5) to centrifuge tube and a small amount of NaN3 to final concentration of 0.02%.
  6. Incubate the tube in hot water (50-60˚c) for 10 minutes to dissolve solids. This can be repeated over several hours, if necessary.
  7. Centrifuge at 5000 x g for 15 minutes to remove any undissolved solids. Store 1ml aliquots in dark microcentrifuge tubes at -20˚C.
  8. Warm tubes to room temperature for use. Opened tubes can be stored at 4˚c for approximately 1 month. Discard if any crystalline material is seen in the tube or on the slides.

Reagents:

  • Mowiol 4-88 (Sigma-Aldrich, cat. no. 81381)
  • Glycerol (Sigma-Aldrich, cat. no. G5516)
  • Trizma Base (Sigma-Aldrich, cat. no. T1503)
  • Sodium Azide (NaN3, Sigma-Aldrich, cat. no. S2002)

#3

Thanks @tberta. This is what we always use daily but I switched to commercial product to save some time. But I don’t think it’s great. I may go back to mowiol.


#4

For a commercial product I like Vectashield (https://vectorlabs.com/vectashield-mounting-medium.html)


#5

Thanks @liz. Do you use this or Mowiol like @tberta?


#6

I routinely use the Vectashield for anything that I will store for longer than a day. For just a quick look when nice pictures are not required I’ll use 50% glycerol in PBS.


#7

I second Vectashield–don’t mistakenly get the hard set, Stick with the stuff Liz included in her first comment–it works pretty well.


#8

Confirm - I tried the hardset thinking it would be advantageous but the slides don’t last nearly as long in storage, becoming discolored and dried out over time.