Mounting worms on slides

Model Organism: Worms Methods & Reagents
1

Hi all,

I have some worms growing on a plate which has Nile Red added to the bacteria. I want to mount these worms on a slide, to look at the Nile Red staining. What’s a good mounting medium to use? Is 70% ethanol okay?

I guess the protocol would be something like this:

  1. wash worms off plate with M9 buffer
  2. spin down buffer + worms
  3. discard supernatant (buffer)
  4. add ~100 uL of 70% ethanol, pipet ethanol + worms onto a slide
  5. cover with coverslip and seal?
2

The standard way to mount worms is in M9 (or another buffer) on an agar (or agarose) pad. See the Cell Biology methods chapter in WormBook for more info:

http://www.wormbook.org/chapters/www_intromethodscellbiology/intromethodscellbiology.html

You need to use a pad so that the coverslip doesn’t squash the animals. And don’t use 70% ethanol; your worms will be very unhappy!

3

I found that EtOH damages my worms.
For short term analysis I usually make 3% agarose pads on microscope slides, add a drop of 1-2mM levamisole add worms and cover with coverslide, you can also seal the coverslide. Good thing about levamisole is that it paralyzes worms while you look at them. Later you can just pick them back onto your plate without any sacrifice.
Hope this helps,
daniel