A kind of platelet transmembrane potential detection method
A technology of transmembrane potential and detection method, which is applied in the field of blood transfusion medicine to achieve the effects of good repeatability of results, improved clinical infusion safety, and low cytotoxicity
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Embodiment 1
[0039] Example 1 Platelet transmembrane potential detection method of the present invention (1)
[0040] 1 Experimental materials and instruments
[0041] Fresh platelets were collected on the same day, put into platelet storage bags, and stored with shaking at 22°C.
[0042] Bis(1,3-dibutylbarbituric acid)trimethineoxonol and DiBAC4(3) (purchased from Sigma) were dissolved in dimethyl sulfoxide to prepare a working solution of 10 μmol / L, and stored at -20°C.
[0043] Gramicidin (Gramicidin, purchased from Sigma, product number 50845) was prepared into a 2 mg / mL working solution using dimethyl sulfoxide and stored at -20°C.
[0044] Hepes-Tyrodes buffer (calcium-free): 137mmol / L sodium chloride, 2.7mmol / L potassium chloride, 12mmol / L sodium bicarbonate, 1mmol / L magnesium chloride, 0.4mmol / L sodium dihydrogen phosphate, 5.5mmol / L glucose , 10mmol / L Hepes, 1mol / L sodium hydroxide to adjust the pH to 7.4.
[0045] Hepes-Tyrodes buffer (high potassium): 2.7mmol / L sodium chlorid...
Embodiment 2
[0056] Example 2 Platelet transmembrane potential detection method of the present invention (2)
[0057] 1 Experimental materials and instruments
[0058] Fresh platelets were collected on the same day, put into platelet storage bags, and stored with shaking at 22°C.
[0059] Bis(1,3-dibutylbarbituric acid)trimethineoxonol and DiBAC4(3) (purchased from Sigma) were dissolved in dimethyl sulfoxide to prepare a working solution of 10 μmol / L, and stored at -20°C.
[0060] Gramicidin (Gramicidin, purchased from Sigma, product number 50845) was prepared into a 2 mg / mL working solution using dimethyl sulfoxide and stored at -20°C.
[0061] Hepes-Tyrodes buffer (calcium-free): 137mmol / L sodium chloride, 1mmol / L potassium chloride, 12mmol / L sodium bicarbonate, 1mmol / L magnesium chloride, 0.4mmol / L sodium dihydrogen phosphate, 5.5mmol / L glucose, 10mmol / L Hepes, 1mol / L sodium hydroxide to adjust the pH to 7.4.
[0062] Hepes-Tyrodes buffer (high potassium): 2mmol / L sodium chloride, 13...
Embodiment 3
[0073] Example 3 Platelet transmembrane potential detection method of the present invention (3)
[0074] 1 Experimental materials and instruments
[0075] Fresh platelets were collected on the same day, put into platelet storage bags, and stored with shaking at 22°C.
[0076]Bis(1,3-dibutylbarbituric acid)trimethineoxonol and DiBAC4(3) (purchased from Sigma) were dissolved in dimethyl sulfoxide to prepare a working solution of 10 μmol / L, and stored at -20°C.
[0077] Gramicidin (Gramicidin, purchased from Sigma, product number 50845) was prepared into a 2 mg / mL working solution using dimethyl sulfoxide and stored at -20°C.
[0078] Hepes-Tyrodes buffer (calcium-free): 134mmol / L sodium chloride, 5mmol / L potassium chloride, 12mmol / L sodium bicarbonate, 1mmol / L magnesium chloride, 0.4mmol / L sodium dihydrogen phosphate, 5.5mmol / L glucose, 10mmol / L Hepes, 1mol / L sodium hydroxide to adjust the pH to 7.4.
[0079] Hepes-Tyrodes buffer (high potassium): 17mmol / L sodium chloride, 12...
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