Biophysical Journal 49: 269-279 (1986)
© 1986 the Biophysical Society

This Article Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Betzig, E. Articles by Kratschmer, E. Search for Related Content PubMed Articles by Betzig, E. Articles by Kratschmer, E.

Near Field Scanning Optical Microscopy (NSOM)

Development and Biophysical Applications

ABSTRACT

A new method for high-resolution imaging, near-field scanning optical microscopy (NSOM), has been developed. The concepts governing this method are discussed, and the technical challenges encountered in constructing a working NSOM instrument are described. Two distinct methods are presented for the fabrication of well-characterized, highly reproducible, subwavelength apertures. A sample one-dimensional scan is provided and compared to the scanning electron micrograph of a test pattern. From this comparison, a resolution of > 1,500 Å (i.e., /3.6) is determined, which represents a significant step towards our eventual goal of 500 Å resolution. Fluorescence has been observed through apertures smaller than 600 Å and signal-to-noise calculations show that fluorescent imaging should be feasible. The application of such imaging is then discussed in reference to specific biological problems. The NSOM method employs nonionizing visible radiation and can be used in air or aqueous environments for nondestructive visualization of functioning biological systems with a resolution comparable to that of scanning electron microscopy.

This article has been cited by other articles:

				E. Betzig and R. J. Chichester Single Molecules Observed by Near-Field Scanning Optical Microscopy Science, November 26, 1993; 262(5138): 1422 - 1425. [Abstract] [PDF]

				E. Betzig and J. K. Trautman Near-Field Optics: Microscopy, Spectroscopy, and Surface Modification Beyond the Diffraction Limit Science, July 10, 1992; 257(5067): 189 - 195. [Abstract] [PDF]

				E. BETZIG, J. K. TRAUTMAN, T. D. HARRIS, J. S. WEINER, and R. L. KOSTELAK Breaking the Diffraction Barrier: Optical Microscopy on a Nanometric Scale Science, March 22, 1991; 251(5000): 1468 - 1470. [Abstract] [PDF]

				K. Lieberman, S. Harush, A. Lewis, and R. Kopelman A Light Source Smaller Than the Optical Wavelength Science, January 5, 1990; 247(4938): 59 - 61. [Abstract] [PDF]