Parvalbumin-tdTomato transgenic whole mouse brain
|Visikol® HISTO-1™||20 mL|
|Visikol® HISTO-2™||20 mL|
Anhydrous (reagent grade)
|Phosphate buffered saline (PBS)||75 mL|
Glass, polypropylene, or polyethylene containers with lids.
Polystyrene is not compatible with Visikol HISTO-2
Warning! Do not treat specimens with other media after clearing. For optimal results, imaging should be performed directly in Visikol HISTO reagents. Visikol HISTO solutions contain anti-fade agents, and specimens cleared with Visikol HISTO do not need to be mounted in an additional anti-fade media. Other solutions (particularly aqueous media) will cloud the tissue or completely reverse the clearing, interfering with 3D volume imaging. Once the cleared tissues have been successfully imaged, see below for additional information on reversing the clearing to perform downstream assays.
For best results, samples should be imaged in Visikol HISTO-2. However, it will damage water dipping objectives present in many OpenSPIM setups. To avoid this, we suggest placing the specimen in a quartz, Simax, or glass cuvette filled with Visikol HISTO-2 then submerging the cuvette in the imaging chamber filled with 2,2’-thiodiethanol, which is compatible with water dipping objectives.LaVision UltraMicroscope II
The UltraMicroscope is directly compatible with Visikol HISTO-2 and requires approximately 150 mL of additional reagent to completely fill the imaging chamber. Visikol HISTO-2 is hygroscopic and imaging quality will degrade in the open imaging chamber for long periods of time.Zeiss Lightsheet Z.1