Background suppression by axially selective activation in single-molecule localization microscopy.
Resolution of a few tens of nanometers has been achieved in fluorescence microscopy with photoswitchable molecules. However, for thick samples, the background brought by the crosstalk of unwanted on-state molecules is nonnegligible. Now we present a background suppression method by using two axial standing waves generated by the interference of two activation beams with the same phases and two deactivation beams with the opposite phases. With spatially incoherent illumination, most activated molecules are located in a thin layer. The performance of such method is simulated with the known photoswitching characteristic of Cy5. With suitable parameters, the thickness of the layer can reach 39 nm (FWHM).
Chen
Danni
D
Key Lab of Optoelectronics Devices and Systems of Ministry of Education/Guangdong Province, Institute of Optoelectronics, Shenzhen University, Shenzhen 518060, China.
Yu
Bin
B
Qu
Junle
J
Niu
Hanben
H