[Histonet] Re: clearing of whole tissue

[aget liang]

thanks, i contact the author, according to Dr.Atsushi Miyawaki, there
should be no salt in the scaLe solution for clearing of tissues. so the
urea is dissolved in miliQ pure water and adjustment of pH is not
necessary, i guess, for urea.

2011/11/19 John Kiernan <jkiernan <@t> uwo.ca>

> The solution described in the paper does not mention a buffer.  Hama et al
> (2011) wrote: "The most effective solution,which we named ScaleA2, was
> composed of 4 M urea, 10% (wt/vol) glycerol and 0.1% (wt/vol) Triton
> X-100."  In the next sentence they said the pH was 7.7.  Later in the paper
> they describe a modified solution suitable for embryos, which has more
> glycerol: "Thus, we developed a modified solution, ScaleU2, composed of 4 M
> urea, 30% glycerol and 0.1% Triton X-100. ScaleU2 requires longer
> incubation times to achieve clearing, but it reduces tissue expansion and
> the fragility of cleared brain samples and is therefore suitable for soft
> samples such as mouse embryos." No mention of pH here.
>
> The paper contains spectacular images.
>
> John Kiernan
> Anatomy, UWO
> London, Canada
>  = = =
> On 18/11/11, *aget liang *<agetliang <@t> gmail.com> wrote:
>
> Did anyone know what is the buffer system for the 4M urea? it is important
> to adjust and maintain pH to 7.7 according to the paper:
> Scale: a chemical approach for fluorescence imaging and reconstruction of
> transparent mouse brain
>
> 2011/11/18 aget liang <agetliang <@t> gmail.com>
>
> > Hi, anyone has experience in tissue clearing? i want to clear spinal cord
> > after fixation. Thanks. YJ
> >
> > --
> > Truth Shall Make You Free
> >
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