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-The new science "CLARITY" enables -
VA-044

"CLARITY" as a way of a new organization transparency was announced by PhD.Karl Deisseroth and his colleagues at Stanford University at the journal Nature in March 2013 . Since CLARITY makes it possible for the immune staining using a fluorescent protein and antibody, it is expected as a useful tool for the analysis of brain circuits.
VA-044 is a reagent used in the transparent process of the paper protocol.

Chung,K et al. : Nature., 497, 332(2013).



1. A mouse brain after CLARITY process

Moue brain

2. Imaging of a mouse brain after CLARITY process

Imaging of a mouse brain after CLARITY process
image

Figure 2. fluorescence observation of Thy1-YFP (H Line) mouse brain after CLARITY process
(A) 3-D observation image from the brain cortex to the hippocampus
(B) dendrites image of the pyramidal cell of the cerebral cortex layer V

Photographing condition: two-photon
Equipment: Olympus FV1000 upright microscope, a 25-fold immersion objective lens

Imaging of a mouse brain after CLARITY process and using antibody
image

Figure 3. fluorescence observation of Thy1-YFP (H Line) mouse brain which has been stained on Iba1 antibody after CLARITY processing (A)  3-D observation image of microglia cells in the brain cortex
(B) an image of observing cortex Ⅰlayer from the surface layer

Shooting conditions: 1 photon
equipment: Olympus FV1000 upright microscope, a 25-fold immersion objective lens



Windows Media format file (3MB) 

Windows Media format file (2MB)



Product Name Package Size Wako catalog No. Storage Condition
VA-044 25 g 223-02112 Keep in a dark place.
100 g 225-02111
500 g 227-02115
VA-044*
Available for sale in the US only
50 g LB-VA044-50GS Keep in a dark place.
500 g 225-02111
Anti Iba1, Rabbit (for Immunocy tochemistry) 50 μg 019-19741 Keep at -20°C.

3. CLARITY Protocol

Protocol

[Hydrogel Monomer Solution composition]
  • VA-044・・・・・・・・・・・・1 g
  • 40% Acrylamide ・・・・・・40 mL
  • 2% Bisacrylamide ・・・・・10 mL
  • 10×PBS ・・・・・・・・・・・40 mL
  • 16% Paraformaldehyde・・・100 mL
  • Saponin・・・・・・・・・・・200 mg
  • Water・・・・・・・・・・・・・・210 mL
[Electrophoresis buffer]
  • Boric acid・・・123.66 g
  • SDS・・・・400 g
  • NaOH ・・・Adjust water to pH 8.5
  • Water・・・・・Measure flask up to 10 L
[Electrophoresis Chamber For Lipid Extraction]
[Circulating pump]

GPU-1, GPU-2 (AZ Science, Japan)

4. CLARITY Reagent List

Product Name Package Size Wako cat. No. Grade Storage Condition
Acrylamide 25 g 017-08012 for Electrophoresis Keep at room temperature.
100 g 019-08011
500 g 011-08015
N,N'-Methylenebis(acrylamide) 25 g 138-06032 for Electrophoresis Keep at 2-10°C.
100 g 130-06031
10×PBS(-) 500 mL 163-25265 for Cell Culture Keep below 25°C.
Paraformaldehyde 100 g 160-16061 for Tissue Fixation Keep at room temperature.
500 g 162-16065
16w/v% Paraformaldehyde Solution, Methanol free 10 mL × 10 A 163-25983 for Electron Microscopy
Saponin, from Soybeans 1 g 198-08853 Wako 1st Grade Keep at 2-10°C.
5 g 192-08851
Boric Acid 100 g 029-02191 JIS Special Grade Keep at room temperature.
500 g 021-02195
4 kg 023-02194
Sodium Dodecyl Sulfate 25 g 190-13982 for Molecular Biology
100 g 192-13981
500 g 194-13985
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