Fluorescence Imaging and Spectroscopy of Biomaterials in Air and Liquid by Scanning Near-Field Optical/Atomic Force Microscopy

Authors

H. Muramatsu, Seiko Instruments Inc.Follow
N. Chiba, Seiko Instruments Inc.
K. Nakajima, Seiko Instruments Inc.
T. Ataka, Seiko Instruments Inc.
M. Fujihira, Tokyo Institute of Technology
J. Hitomi, Seiko Instruments Inc.
T. Ushiki, Niigata University School of Medicine

Abstract

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.

Recommended Citation

Muramatsu, H.; Chiba, N.; Nakajima, K.; Ataka, T.; Fujihira, M.; Hitomi, J.; and Ushiki, T. (1996) "Fluorescence Imaging and Spectroscopy of Biomaterials in Air and Liquid by Scanning Near-Field Optical/Atomic Force Microscopy," Scanning Microscopy: Vol. 10: No. 4, Article 6.
Available at: https://digitalcommons.usu.edu/microscopy/vol10/iss4/6