CLARITY of Skeletal Muscle

kbabos

We will be using CLARITY to image mouse skeletal muscle, but we are wondering if anyone has tried CLARITY on any muscle samples? We are aware that the cartilage and tendons in the muscle might prove tricky to clear with CLARITY, but we're hoping to find out if anyone has had success with any sort of muscle to-date. Also, if successful, are there any tricks with the process that you can recommend we include in our protocol?
Many thanks!
Kim

Fleck

Hi, did you already proceed? I plan to clear a VNO which is encapsulated in cartilage. It will also be possible to remove the cartilage but then the organ will be very fragile! Are there some ideas what would be the best approach.

KatherineHolzem

@kbabos. We have been working with heart muscle, and the clarity procedure does work. However, in our hands, muscle takes much longer than neural tissue to clear (presumably due to the higher ratio of ECM to lipids). We are trying many adaptations to the original procedure to make the process faster, including using bis-free gel as suggested on this forum. It is difficult to put muscle through the original ETC process long enough to clear without damage.

That being said, we have some clear tissue and fluorescence labeling. We have been able to image through a whole mouse heart and stain for various markers. The ECM also presents a problem with staining though because there is a lot of autofluorescence... particularly in the green and red wavelength regions.

helen

@KatherineHolzem. I have been clearing brain tissue for the last couple of months, but am about to start working on hearts as part of a collaborative project. Could you tell me the settings you use for your heart muscle protocol? Thanks, Helen

neuro100

@KatherineHolzem, I'm also interested in your muscle protocol, if you'd be willing to share it! I'd be grateful to not have to re-invent the wheel. NB we have no plans to work on heart.