Fast two-dimensional standing-wave total-internal-reflection fluorescence microscopy using acousto-optic deflectors.
We report a scheme for 2D standing-wave total-internal-reflection fluorescence microscopy. Standing-wave patterns are generated by two interfering beams coupled through the objective lens. Period, angular orientation, and phase of standing waves are controlled entirely by acousto-optic deflectors. The lateral resolution improvement of 100 nm is combined with an axial selectivity of <100 nm by utilizing an evanescent standing-wave pattern. This technique can provide real-time imaging of subresolution structures in live biological specimens near a glass-water interface.
Gliko
Olga
O
Department of Otorhinolaryngology-Head and Neck Surgery, Baylor College of Medicine, Houston, TX 77030, USA.
Brownell
William E
WE
Saggau
Peter
P
D000162:Acoustics / Q000295:instrumentation*; D017076:Computer-Aided Design; D004867:Equipment Design; D019544:Equipment Failure Analysis; D007909:Lenses; D008029:Lighting / Q000295:instrumentation* / Q000379:methods; D008856:Microscopy, Fluorescence / Q000295:instrumentation* / Q000379:methods; D012031:Refractometry / Q000295:instrumentation*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity