1：Experimental Design and Method Selection:

The study used blinking QDs with emission at 605 nm as a study model and developed a method to statistically evaluate the fluorescence intensity of a large number of single blinking QDs using a confocal fluorescence microscope. Gaussian distribution was used to evaluate the fluorescence intensity distribution.

2：Sample Selection and Data Sources:

Qdot? 605 ITK? amino [PEG] CdSe quantum dots were used. The samples were prepared by mixing QDs with PBS and placing them on plasma-rinsed glass coverslips for fluorescence imaging.

3：List of Experimental Equipment and Materials:

Coverslips from Fisher Scientific, Qdot? 605 ITK? amino [PEG] CdSe quantum dots from Invitrogen, Hitachi H-7000FA TEM, Andor Revolution XD spinning-disk confocal microscope equipped with EMCCD (Andor iXon ultra897), 561 nm laser (DPSS, USA), 605/20 nm filter (Chroma).

4：Experimental Procedures and Operational Workflow:

QDs were imaged using TEM and confocal fluorescence microscopy. The intensity versus time plots of QD spots were analyzed by Image J. Statistical intensity analysis was performed by Image-Pro Plus (IPP) using Gaussian distribution.

5：Data Analysis Methods:

The fluorescence intensity distribution of QDs was evaluated using Gaussian distribution. The dispersion degree of intensity data was characterized with standard deviation (SD).