Optical trapping and manipulation of live T cells with a low numerical aperture lens.

An optical manipulation system that employs both optical and temperature gradients to simultaneously enable trapping, manipulating and imaging of live cells with a low magnification, low numerical aperture objective lens (10x/0.4 N.A.) is reported. This approach negates the requirement for a high N.A. lens used in traditional optical trapping. Our system comprised a dual scanning system and two independent lasers which provided the ability to move the trapping spot independently of the confocal imaging process in close to real-time and without pre-programming. To demonstrate the efficacy of this innovative method, trapping and manipulation of live T cells was simultaneously performed over a field of view exceeding 1 mm(2) for extended periods without compromising cell viability.

[1]  Daniel Day,et al.  A microfluidic refractive index sensor based on an integrated three-dimensional photonic crystal , 2008 .

[2]  M Mazilu,et al.  Dual beam fibre trap for Raman micro-spectroscopy of single cells. , 2006, Optics express.

[3]  D. J. Segelstein The complex refractive index of water , 1981 .

[4]  L. Goldstein,et al.  Bead movement by single kinesin molecules studied with optical tweezers , 1990, Nature.

[5]  K. Dholakia,et al.  Microfluidic sorting in an optical lattice , 2003, Nature.

[6]  A. B. Lyons,et al.  Determination of lymphocyte division by flow cytometry. , 1994, Journal of immunological methods.

[7]  A. Ashkin Forces of a single-beam gradient laser trap on a dielectric sphere in the ray optics regime. , 1992, Methods in cell biology.

[8]  P. Matsudaira,et al.  Laser-guided assembly of heterotypic three-dimensional living cell microarrays. , 2006, Biophysical journal.

[9]  C. Wandrey,et al.  Purification of Polymeric Biomaterials , 2001, Annals of the New York Academy of Sciences.

[10]  A. F. Marée,et al.  Spatial modelling of brief and long interactions between T cells and dendritic cells , 2007, Immunology and cell biology.

[11]  Ignacio Tinoco,et al.  Temperature control methods in a laser tweezers system. , 2005, Biophysical journal.

[12]  W. Faulk,et al.  Propidium iodide as a nuclear marker in immunofluorescence. II. Use with cellular identification and viability studies. , 1981, Journal of immunological methods.

[13]  Max Born,et al.  Principles of optics - electromagnetic theory of propagation, interference and diffraction of light (7. ed.) , 1999 .

[14]  S W Hell,et al.  Heating by absorption in the focus of an objective lens. , 1998, Optics letters.

[15]  Stefan Schinkinger,et al.  Müller cells are living optical fibers in the vertebrate retina , 2007, Proceedings of the National Academy of Sciences.

[16]  W Sibbett,et al.  Creation and Manipulation of Three-Dimensional Optically Trapped Structures , 2002, Science.

[17]  Kort Travis,et al.  Fluorescence ratio thermometry in a microfluidic dual-beam laser trap. , 2007, Optics express.

[18]  M W Berns,et al.  Parametric study of the forces on microspheres held by optical tweezers. , 1994, Applied optics.

[19]  Jesper Gluckstad,et al.  Fully dynamic multiple-beam optical tweezers. , 2002, Optics express.

[20]  X. Gan,et al.  Trapping force by a high numerical-aperture microscope objective obeying the sine condition , 1997 .