Clearing protocole for Lightsheet microscopy

Probes & Prep
1

Hello,
I read some article, and I found a lot of different protocol for the clearing.

We already use some protocol in our platform with a lightsheet Z1 from Zeiss.

  • X clarity for adult mice brain
  • CUBIC for organoid and embryo mice brain
  • RapiClear +/- RIMS for small samples (with +/- result)
  • Reverse BABB (an organic solvent based method) for lungs. Careful organic solvent properties can be very corrosive for the lens. This is why We reverse a bit the protocol.

Do you have some other protocol to share with me based on your experiment?

2

Are you looking for more of a review of clearing techniques?

One of our users is trying FlyClear

3

CubicL and Ethyl Cinnamate work very well an a large variety of samples…

4

This article from the Chung lab was suggested to me while we were demoing a Z1: https://static1.squarespace.com/static/561beb9ae4b0edc5a7075cf2/t/57ba20d69f7456e36f2f6ce3/1504722155038/Chung+lab_CLARITY+protocol_08192016.pdf
It has some interesting cost breakdowns, among other things.

5

Dear Mix,
yes I looking for new protocol. If possible between RI 1.33 and RI 1.45 or 1.49
Thank you for this protocol.

6

Hello,
We have in the plateform a Z1 from Zeiss and I’m worried about Ethyl Cinnamate because RI will be 1.56
My microscope is set to work with 1.33 or 1.45. I already try the CUBIC 2 with success +/- RI 1.48
Do you have some advice to reduce RI with Ethyl Cinnamate?

7

Thank your for your answer. We already use the X-Clarity protocol with success on brain but this protocol is not very good for small samples and lungs.