. 2019 Jun 10;9:8374. doi: 10.1038/s41598-019-44897-7

Table 2.

Summarized inCLARITY protocol.

	Fixation	Hydrogel polymerization	Lipid-clearing	Wash	IHC	Wash	Mounting
Procedure	Transcardial PFA 4%-fixation	Acrylamide/Bisacrilamide + V-50 in 4% PFA in PBS, pH 7.4	8% SDS, pH 7.4	PBST 0.1%, pH 7.4	Primary & secondary antibodies in PBST 0.1%	PBST 0.1%, pH 7.4	Mowiol
Time	<1 h Post-fixation PFA 4% Overnight 4 °C	2 h at 4 °C followed by 4 h at 37 °C	7–20 days at RT	1 day at RT	5–8 days at RT	1 day at RT	Until dry

Fixation

Hydrogel polymerization

Lipid-clearing

Wash

IHC

Wash

Mounting

Procedure

Transcardial PFA 4%-fixation

Acrylamide/Bisacrilamide + V-50 in 4% PFA in PBS, pH 7.4

8% SDS, pH 7.4

PBST 0.1%, pH 7.4

Primary & secondary antibodies in PBST 0.1%

PBST 0.1%, pH 7.4

Mowiol

Time

<1 h

Post-fixation PFA 4% Overnight 4 °C

2 h at 4 °C followed by 4 h at 37 °C

7–20 days at RT

1 day at RT

5–8 days at RT

1 day at RT

Until dry