Lavision biotek and Zeiss now have commercially available light sheet systems. The approach looks interesting because when combined with clearing agents, and fluorescently labelled structures it allows very large specimens to be imaged. So for instance using the lavision system an entire mouse brain could be imaged (assuming that it is a GM animal with GFP tagged molecule) without the need to section it. It comes at the expense of a lower resolution than what would be achievable if the specimen was sectioned. Has anybody out there had experiences good or bad with this technology?
I have recently started using this technology to image lungs from mice and currently, I am optimizing the use of fluorescent labelled antibodies, as well as transfer of cells tagged with fluorescent dye prior to chemical clearing. I have had both good and bad experiences, but otherwise it is a great technology.
Hello Fredrerick,
You can use the "traditionell" clearing agents (Murrays clear): EtOH and BABB (1part benzylalcohol plus 2 parts benzylbenzoat)/ peroxidfree.
Or
THF/DBE- Clearing: Tetrahydrofuran/Dibenzylether (Becker & Jährling et al. 2011)http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0033916
This clearing is also called: "3Disco-Clearing".
Be careful with the peroxides; they should be removed before the experiment....
Best wishes from Vienna,
Nina
Can light sheet microscopy be used for imaging the brain of anesthetized mice repeatedly?
Dear Carmel,
The simple answer is no. The real benefit of LSM is when you remove remove the lipids from your specimen, which when combined with light sheet scanning and refractive index matching medium, will allow light to penetrate deeper into the tissue and allow more to come back to the detector, thus allowing you to image larger structures. You can't clear living tissue (at least at this stage, as far as I'm aware).
Rohan.
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