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SWITCH (system-wide control of interaction time and kinetics of chemicals) is a simple, scalable, and generalizable tissue-processing method for proteomic imaging of intact biological systems. The SWITCH tissue-processing and clearing method provides access to high-dimensional multi-scale information that may help to understand health and disease from molecules to cells to entire systems.
Developed by the Chung Lab.
|32% Paraformaldehyde (PFA)||Electron Microscopy Sciences, 15714-S|
|50% Glutaraldehyde (GA)||Electron Microscopy Sciences, 16310|
|Potassium hydrogen phthalate||Sigma-Aldrich, P1088|
|Sodium azide||Sigma-Aldrich, S2002|
|Triton-X (TX) 100||Amresco, 0694|
|Sodium dodecyl sulfate (SDS)||Sigma-Aldrich, L3771|
|Sodium sulfite||Sigma-Aldrich, S0505|
|N- methyl-D-glucamine||Sigma-Aldrich, M2004|
|Diatrizoic acid||Sigma-Aldrich, D9268|
|60% iodixanol||Sigma-Aldrich, D1556|
This solution should be made fresh immediately prior to performing perfusion and kept on ice at all times. It is recommended to chill all of the separate ingredients before mixing the components.
|10X PBS||4 mL|
|32% PFA||5 mL|
|50% GA||0.8 mL|
Titrate a bottle of PBS to pH 3 using HCl. Create solutions of 0.1 M HCl in water and 0.1 M potassium hydrogen phthalate (KHP) in water. Finally, mix these solutions in a ratio of 2:1:1 (pH 3 PBS):(0.1 M HCl):(0.1 M KHP).
To this solution, add a stock solution of GA to make a final concentration of 4% GA. Ensure that this solution stays cold at all times. It is recommended to chill the solution before adding GA.
Add a stock solution of GA to PBS (pH 7.4) to make a final concentration of 1% GA. Ensure that this solution stays cold at all times. It is recommended to chill the PBS before adding GA.
To PBS, add TX-100 to a final concentration of 0.1% (v/v). Also, add sodium azide to a final concentration of 0.02% (w/v).
|47% Iodixanol solution in water*||10 mL|
|Diatrizoic acid||4.24 g|