We have developed scanning near-field optical/atomic force microscopy (SNOM/AFM). The SNOM/AFM uses a bent optical fiber simultaneously as a dynamic force AFM cantilever and a SNOM probe. Resonant frequency of the optical fiber cantilever is 15-40 kHz. Optical resolution of the SNOM/AFM images shows less than 50 nm. The SNOM/ AFM system contains photon counting system and polychrometer/intensified coupled charge devise (ICCD) system to observe fluorescence image and spectrograph of micro areas, respectively. Cultured cells were stained with fluorescein isothiocyanate (FITC)-labeled anti-keratin antibody or FITC-labeled phalloidin after treatment with Triton X-100. Fluorescence and topographic images were obtained in air and water. The fluorescence images showed clear images of keratin and actin filaments. The SNOM/AFM is perfect to observe biomaterials in liquid with a liquid chamber while the topographic Images showed subcellular structures which correspond to keratin and actin filaments.
Muramatsu, H.; Chiba, N.; Nakajima, K.; Ataka, T.; Fujihira, M.; Hitomi, J.; and Ushiki, T.
"Fluorescence Imaging and Spectroscopy of Biomaterials in Air and Liquid by Scanning Near-Field Optical/Atomic Force Microscopy,"
Scanning Microscopy: Vol. 10
, Article 6.
Available at: https://digitalcommons.usu.edu/microscopy/vol10/iss4/6