Teaching PAINT

AUTHORS: Albertazzi L.

Introductory Information

The shared data consist in a single single-molecule localization imaging acquisition of binding sites exposed on the surface of streptavidin-coated polystyrene particles. Data were obtained with DNA Points Accumulation for Imaging in Nanoscale Topography (DNA-PAINT) procedure by loading streptavidins with biotinylated DNA docking strands and then introducing a dye-conjugated DNA imager strand in solution for single-molecule localization. 

The acquisition is short (500 frames) to facilitate data loading and analysis by students during teaching sessions on data analysis. 

Data specific information

streptavidin-coated polystyrene particles (320 nm diameter) incubated with biotin-conjugated DNA docking strand in excess (10 M) at different concentrations; washed and imaged on glass slide in imaging buffer (5 Mm Tris-HCl, 10 mM MgCl2, 1mM EDTA, 0.05% Tween-20, pH = 8) containing complementary DNA imager strand conjugated with Atto647N at 0.1 nM.

Acquisition parameters:
256x256 pixels region of camera, pixel size = 160nm
total internal reflection irradiation
main excitation = 647 nm (50% of max power)
reference excitation = 561 nm (0.5% of max power), 1 frame every 100 of main-channel
camera exposure time = 50ms (20 Hz)
500 frames in main channel
Max power 647 nm laser = 160 mW
Max power 561 nm laser = 80 mW
Camera baseline value = 100
