Fig. 1. Principle of tagging strategy

Fig. 2. Image changes before and after buffer treatment, taking microtubules as an example, stained with Alexa488. (a) Image without buffer; (b) image after adding buffer; (c) image brightness before and after buffer treatment; (d) anti-quenching ability of fluorescence signal before and after buffer treatment; (e) fluctuation amplitude of fluorescence signal before and after buffer treatment

Fig. 3. Changes of image on-time ratio before and after buffer treatment. (a) Distribution of image on-time ratio before adding buffer; (b) statistics of image on-time ratio before adding buffer; (c) distribution of image on-time ratio after adding buffer; (d) statistics of image on-time ratio after adding buffer

Fig. 4. SOFI reconstruction images of microtubules before and after buffer treatment. (a) Original images of microtubules without buffer treatment and the first-order, second-order, third-order, and fourth-order images after SOFI processing; (b) brightness distribution of underlined part in figure (a); (c) original image of microtubules after adding buffer and the first-order, second-order, third-order, and fourth-order images after SOFI processing; (d) brightness distribution of underlined part in figure (c)

Fig. 5. SOFI reconstruction images of vesicles before and after buffer treatment. (a) Original images of vesicles without buffer and the first-order, second-order, third-order, and fourth-order images after SOFI treatment; (b) brightness distribution of underlined part in figure (a); (c) original images of vesicle after adding buffer and the first-order, second-order, third-order, and fourth-order image after SOFI treatment; (d) brightness distribution of underlined part in figure (c)