Abstract

Reflection interference contrast microscopy (RICM) is a technique for measuring the shape and position of microscopic objects in solution; it has many biological and biophysical applications. Use of RICM for long-time acquisitions requires minimizing defocusing effects that are due to thermal and mechanical drift. We present a simple stabilizing method that accomplishes this using an image-analysis-based linear focus function to establish feedback control of the focal position. While implementing this routine, we used RICM for independent measurement of the apparent fluctuation in the vertical position of an immobilized bead: the measured height had a standard deviation of $0.12\mathrm{nm}$ during a 45 min acquisition while under feedback control, demonstrating the high stability achievable with our approach.

© 2008 Optical Society of America

Reflection interference contrast microscopy of arbitrary convex surfaces

Jose C. Contreras-Naranjo, James A. Silas, and Victor M. Ugaz
Appl. Opt. 49(19) 3701-3712 (2010)

Microinterferometry: three-dimensional reconstruction of surface microtopography for thin-film and wetting studies by reflection interference contrast microscopy (RICM)

Gerald Wiegand, Klaus R. Neumaier, and Erich Sackmann
Appl. Opt. 37(29) 6892-6905 (1998)

Influence of surface reflective properties on differential interference contrast microscopy

Olivier Theodoly, Sylvain Gabriele, and Marie-Pierre Valignat
Opt. Express 16(7) 4547-4558 (2008)

Quantitative phase and refractive index analysis of optical fibers using differential interference contrast microscopy

Betty Kouskousis, Daniel J. Kitcher, Stephen Collins, Ann Roberts, and Greg W. Baxter
Appl. Opt. 47(28) 5182-5189 (2008)

Hyperspectral Fourier transform spectrometer for reflection spectroscopy and spectral self-interference fluorescence microscopy

Julia Rentz Dupuis, James Needham, Emre Özkumur, David A. Bergstein, Bennett B. Goldberg, James R. Engel, David L. Carlson, and M. Selim Ünlü
Appl. Opt. 47(9) 1223-1234 (2008)