Mounting for imaging

Jalal

After incubating in focus clear, the protocol says to mount in a chamber covered with a Wllco dish. Has anyone tried using simple coverslips?

We have Corning 1 1/2 cover slips that are about the same thickness of the glass bottoms of the Willco dish (~.17mm). May give them a try, but maybe the refractive properties are different.

If anyone has come up with a fun mounting solution that works, I'd be happy to hear it!

gjones

My understanding was that you only need to use Wilco dish if you're going to image with an immersion lens because it's just a well with a coverslip as the base. If you're going to use a dry lens then you could certainly just use regular coverslips.

nicolasrenier

I use regular coverslips with blu-tack and qwik-sil (the same protocol described online, but with regular glass instead of the super expensive wilco. I don't understand your comment @gjones : you can also use immersion objectives with a regular coverslip, even water immersion.

@Jalal : did you get a sample worthy of imaging? I would be very interested to know what kind of fluorophore you're about to image!

gjones

I meant for immersion objectives with a large working distance, i.e. 1-8mm.

Jalal

@nicholasrenier : no imaging yet but I'll post some pictures of embryos that I've attempted to clear. Looks promising i.e. they look more transparent than uncleared embryos, but sadly I cannot read Ramon y Cajal through them yet

David

I invite you to consult that link for your imaging set up:
http://www.gracebio.com/life-science-products

aysegul

The page we requested was not found @David ?

Clara

Hi! has someone tried to replicate the mounting technique described in Poguzhelskaya paper on 2014? they simply put the tissue slices (1 or 1,5 mm thick) on a glass slide and used a regular coverslip, no mounting media involved, just some drops of PBST to avoid drying. I was wondering about trying this instead of using 85% glycerol.... any thoughts?

MariaSuessmilch

Hey Clara, I am wondering whether you have tried this method yet and if so, did you have any luck with it?