ScaleS clearing and multiphoton

[Phillips, Thomas E.]

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Can anyone using ScaleS as a clearing medium/protocol comment on what objectives and immersion fluids they are using and how deep they can image? We have Leica confocals and they don't sell an objective designed for Scale like Olympus and Zeiss do. Is the RI of ScaleS the same as ScaleA2 or is it higher? Tom Phillips

[Olaf Selchow]

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Hi Tom,

As mentioned in the paper (Table on page 9)
Hama, H. et al. ScaleS: an optical clearing palette for biological imaging. Nat Neurosci (2015). doi:10.1038/nn.4107
the RI of Scale S should be 1.44 ... so comparable to CLARTIY and the like. It is different (!) from ScaleA2 (1.38).
I remember that ScaleS can be imaged quite well with the Zeiss 20× objective lens (NA = 1.0, WD = 5.6 mm, RI = 1.45).
Penetration depth in brain tissue was several millimetres and absolutely comparable to CLARITY or CUBIC. I have not see other tissue types cleared with ScaleS though.
I guess the Olympus and Leica lenses would work just as well.
Note that the spectral properties of CLARITY, Scale and other methods are not the same --> keep this in mind when doing multi-color / coloc.

Hope this helps,
Olaf

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Olaf Selchow
Microscopy & BioImaging Consultant
[hidden email]