| Characterization of Photodamage to Escherichia coli in Optical Traps Biophysical Journal, Volume 77, Issue 5, 1 November 1999, Pages 2856-2863 Keir C. Neuman, Edmund H. Chadd, Grace F. Liou, Keren Bergman and Steven M. Block Abstract Optical tweezers (infrared laser-based optical traps) have emerged as a powerful tool in molecular and cell biology. However, their usefulness has been limited, particularly in vivo, by the potential for damage to specimens resulting from the trapping laser. Relatively little is known about the origin of this phenomenon. Here we employed a wavelength-tunable optical trap in which the microscope objective transmission was fully characterized throughout the near infrared, in conjunction with a sensitive, rotating bacterial cell assay. Single cells of were tethered to a glass coverslip by means of a single flagellum: such cells rotate at rates proportional to their transmembrane proton potential (Manson et al., 1980. 138:541–561). Monitoring the rotation rates of cells subjected to laser illumination permits a rapid and quantitative measure of their metabolic state. Employing this assay, we characterized photodamage throughout the near-infrared region favored for optical trapping (790–1064nm). The action spectrum for photodamage exhibits minima at 830 and 970nm, and maxima at 870 and 930nm. Damage was reduced to background levels under anaerobic conditions, implicating oxygen in the photodamage pathway. The intensity dependence for photodamage was linear, supporting a single-photon process. These findings may help guide the selection of lasers and experimental protocols best suited for optical trapping work. Abstract | Full Text | PDF (281 kb) |
| Laser-Guided Assembly of Heterotypic Three-Dimensional Living Cell Microarrays Biophysical Journal, Volume 91, Issue 9, 1 November 2006, Pages 3465-3473 G.M. Akselrod, W. Timp, U. Mirsaidov, Q. Zhao, C. Li, R. Timp, K. Timp, P. Matsudaira and G. Timp Abstract We have assembled three-dimensional heterotypic networks of living cells in hydrogel without loss of viability using arrays of time-multiplexed, holographic optical traps. The hierarchical control of the cell positions is achieved with, to our knowledge, unprecedented submicron precision, resulting in arrays with an intercell separation <400nm. In particular, we have assembled networks of Swiss 3T3 fibroblasts surrounded by a ring of bacteria. We have also demonstrated the ability to manipulate hundreds of simultaneously into two- and three-dimensional arrays with a time-averaged power <2mW per trap. This is the first time to our knowledge that living cell arrays of such complexity have been synthesized, and it represents a milestone in synthetic biology and tissue engineering. Abstract | Full Text | PDF (686 kb) |
| An Automated Two-Dimensional Optical Force Clamp for Single Molecule Studies Biophysical Journal, Volume 83, Issue 1, 1 July 2002, Pages 491-501 Matthew J. Lang, Charles L. Asbury, Joshua W. Shaevitz and Steven M. Block Abstract We constructed a next-generation optical trapping instrument to study the motility of single motor proteins, such as kinesin moving along a microtubule. The instrument can be operated as a two-dimensional force clamp, applying loads of fixed magnitude and direction to motor-coated microscopic beads moving in vitro. Flexibility and automation in experimental design are achieved by computer control of both the trap position, via acousto-optic deflectors, and the sample position, using a three-dimensional piezo stage. Each measurement is preceded by an initialization sequence, which includes adjustment of bead height relative to the coverslip using a variant of optical force microscopy (to ±4nm), a two-dimensional raster scan to calibrate position detector response, and adjustment of bead lateral position relative to the microtubule substrate (to ±3nm). During motor-driven movement, both the trap and stage are moved dynamically to apply constant force while keeping the trapped bead within the calibrated range of the detector. We present details of force clamp operation and preliminary data showing kinesin motor movement subject to diagonal and forward loads. Abstract | Full Text | PDF (528 kb) |
Copyright © 1996 The Biophysical Society. All rights reserved.
Biophysical Journal, Volume 70, Issue 3, 1529-1533, 1 March 1996
doi:10.1016/S0006-3495(96)79716-3
Research Article
H. Liang, K.T. Vu, P. Krishnan, T.C. Trang, D. Shin, S. Kimel and M.W. Berns
Beckman Laser Institute and Medical Clinic, University of California at Irvine 92715, USA.
A study on clonal growth in Chinese hamster ovary (CHO) cells was conducted after exposure to optical trapping wavelengths using Nd:YAG (1064 nm) and tunable titanium-sapphire (700–990 nm) laser microbeam optical traps. The nuclei of cells were exposed to optical trapping forces at various wavelengths, power densities, and durations of exposure. Clonal growth generally decreased as the power density and the duration of laser exposure increased. A wavelength dependence of clonal growth was observed, with maximum clonability at 950–990 nm and least clonability at 740–760 nm and 900 nm. Moreover, the most commonly used trapping wavelength, 1064 nm from the Nd:YAG laser, strongly reduced clonability, depending upon the power density and exposure time. The present study demonstrates that a variety of optical parameters must be considered when applying optical traps to the study of biological problems, especially when survival and viability are important factors. The ability of the optical trap to alter either the structure or biochemistry of the process being probed with the trapping beam must be seriously considered when interpreting experimental results.