BSA-based super-resolution imaging probe as well as preparation method and application thereof
A super-resolution imaging and probe technology, applied in the field of super-resolution imaging probes, can solve the problems of complex imaging process, troublesome, and limited super-resolution imaging applications, and achieve the effect of long blinking time and good photostability
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Embodiment 1
[0038] Dissolve 2 mg of BSA in 200 μL of PBS solution, adjust the pH to 8.3 with 0.5 M sodium bicarbonate, then add 2 μL of 1 mg / mL Alexa Fluor 594 NHS ester to the solution and react on a shaking table at room temperature for 1 hour to obtain a solution It is the BSA solution coupled with Alexa Fluor 594.
[0039]Dissolve 20 mg of BSA in 2 mL of PBS solution, and then add 200 μL of the BSA solution coupled with AlexaFluor 594 prepared above to obtain a mixed BSA solution (total concentration of BSA is 10 mg / mL). Stir and continue to add 2 mL of absolute ethanol, then add 50 μL of 8% (v / v) glutaraldehyde solution, and stir for 12 h (final concentration of glutaraldehyde is 0.18%) to obtain dye-incorporated Finally, the sample was purified 5 times with a 50 KD ultrafiltration tube at 6500 round / min for 20 min to obtain BSA nanospheres doped with Alexa Fluor 594. Subsequently, take 500 μL of BSA nanosphere solution mixed with Alexa Fluor594, add 50 μL of 10% (v / v) glutaraldehyd...
Embodiment 2
[0042] Dissolve 5 mg of BSA in 200 μL PBS solution, adjust the pH to 8.3 with 0.5 M sodium bicarbonate, then add 5 μL of 1 mg / mL Alexa Fluor 594-NHS easter to the solution and react in a shaker at room temperature for 1 hour, and the obtained solution is BSA solution conjugated with Alexa Fluor 594.
[0043] Dissolve 20 mg of BSA in 2 mL of PBS solution, and then add 200 μL of the BSA solution coupled with AlexaFluor 594 prepared above to obtain a mixed BSA solution (total BSA concentration 11.4 mg / mL). Stir and continue to add 2 mL of absolute ethanol, then add 50 μL of 8% (v / v) glutaraldehyde solution, and stir for 18 hours (the final concentration of glutaraldehyde is 0.18%) to obtain the dye-incorporated BSA nanosphere solution, and finally the sample was purified 5 times with a 50KD ultrafiltration tube at 6500 round / min for 20 minutes to obtain BSA nanospheres mixed with Alexa Fluor 594. Then, take 500 μL of BSA nanosphere solution mixed with AlexaFluor 594, add 50 μL o...
Embodiment 3
[0045] Dissolve 2 mg of BSA in 200 μL of PBS solution, adjust the pH to 8.5 with 0.5 M sodium bicarbonate, then add 2 μL of 1 mg / mL Alexa Fluor 594-NHS easter to the solution and react in a shaker at room temperature for 1 hour, and the obtained solution is A solution in BSA conjugated with Alexa Fluor 594.
[0046] Dissolve 20 mg of BSA in 2 mL of PBS solution, and then add 200 μL of the BSA solution coupled with AlexaFluor 594 prepared above to obtain a mixed BSA solution (total concentration of BSA is 10 mg / mL). Stir and continue to add 4 mL of absolute ethanol, then add 50 μL of 8% (v / v) glutaraldehyde solution, and stir for 16 hours (the final concentration of glutaraldehyde is 0.18%) to obtain BSA nanoparticles mixed with dye. sphere solution, and finally the sample was purified 5 times with a 50KD ultrafiltration tube at 6500 round / min for 20 minutes to obtain BSA nanospheres doped with Alexa Fluor 594. Subsequently, take 500 μL of the BSA nanosphere solution mixed wit...
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