Two-point separation in far-field super-resolution fluorescence microscopy based on two-color fluorescence dip spectroscopy, Part I: Experimental evaluation.
The two-point resolution of a novel two-color far-field super-resolution fluorescence microscopy was evaluated by measuring fluorescent beads 100 nm in diameter. This microscopy is based on a combination of two-color fluorescence dip spectroscopy and a phase-modulation technique for a laser beam. By simply introducing two-color laser light, the size of the fluorescent image of a bead was shrunk down to a diameter of 250 nm from the diffraction-limited image with a diameter of 360 nm. For two closely adjacent fluorescent beads with a separation distance of 350 nm, the two-color microscope clearly gave separated fluorescence images, while the conventional one-color fluorescence microscope could not resolve them. It has been proved that our technique breaks Rayleigh's diffraction limit.
Watanabe
Takeshi
T
Tokyo Institute of Technology, Chemical Resources Laboratory, 4259 Nagatsuta-cho, Midoriku, Yokohama 226-8503 Japan.
Iketaki
Yoshinori
Y
Omatsu
Takashige
T
Yamamoto
Kimihisa
K
Fujii
Masaaki
M
D005456:Fluorescent Dyes; D007089:Image Enhancement / Q000295:instrumentation / Q000379:methods*; D007834:Lasers; D036641:Microscopy, Fluorescence, Multiphoton / Q000295:instrumentation* / Q000379:methods*; D043942:Nanotubes / Q000032:analysis*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
D005456:Fluorescent Dyes