Manipulation of Suspended Single Cells by Microfluidics and Optical Tweezers.

Chondrocytes and osteoblasts experience multiple stresses in vivo. The optimum mechanical conditions for cell health are not fully understood. This paper describes the optical and microfluidic mechanical manipulation of single suspended cells enabled by the µPIVOT, an integrated micron resolution particle image velocimeter (µPIV) and dual optical tweezers instrument (OT). In this study, we examine the viability and trap stiffness of cartilage cells, identify the maximum fluid-induced stresses possible in uniform and extensional flows, and compare the deformation characteristics of bone and muscle cells. These results indicate cell photodamage of chondrocytes is negligible for at least 20 min for laser powers below 30 mW, a dead cell presents less resistance to internal organelle rearrangement and deforms globally more than a viable cell, the maximum fluid-induced shear stresses are limited to ~15 mPa for uniform flows but may exceed 1 Pa for extensional flows, and osteoblasts show no deformation for shear stresses up to 250 mPa while myoblasts are more easily deformed and exhibit a modulated response to increasing stress. This suggests that global and/or local stresses can be applied to single cells without physical contact. Coupled with microfluidic sensors, these manipulations may provide unique methods to explore single cell biomechanics.

Mechanical stress analysis of microfluidic environments designed for isolated biological cell investigations.

Advancements in technologies for assessing biomechanics at the cellular level have led to discoveries in mechanotransduction and the investigation of cell mechanics as a biomarker for disease. With the recent development of an integrated optical tweezer with micron resolution particle image velocimetry, the opportunity to apply controlled multiaxial stresses to suspended single cells is available (Neve, N., Lingwood, J. K., Zimmerman, J., Kohles, S. S., and Tretheway, D. C., 2008, "The muPIVOT: An Integrated Particle Image Velocimetry and Optical Tweezers Instrument for Microenvironment Investigations," Meas. Sci. Technol., 19(9), pp. 095403). A stress analysis was applied to experimental and theoretical flow velocity gradients of suspended cell-sized polystyrene microspheres demonstrating the relevant geometry of nonadhered spherical cells, as observed for osteoblasts, chondrocytes, and fibroblasts. Three flow conditions were assessed: a uniform flow field generated by moving the fluid sample with an automated translation stage, a gravity driven flow through a straight microchannel, and a gravity driven flow through a microchannel cross junction. The analysis showed that fluid-induced stresses on suspended cells (hydrodynamic shear, normal, and principal stresses in the range of 0.02-0.04 Pa) are generally at least an order of magnitude lower than adhered single cell studies for uniform and straight microchannel flows (0.5-1.0 Pa). In addition, hydrostatic pressures dominate (1-100 Pa) over hydrodynamic stresses. However, in a cross junction configuration, orders of magnitude larger hydrodynamic stresses are possible without the influence of physical contact and with minimal laser trapping power.