Fluorimetric multiparameter cell assay at the single cell level fabricated by optical tweezers

A fluorimetric multi‐parameter cell sensor at the single cell level is presented which makes it possible to observe the physiological behavior of different cell lines, different physiological parameters, and statistical data at the same time. Different cell types were immobilized at predefined positions with high accuracy using optical tweezers and adhesion promoting surface layers. The process is applicable to both adherent and non‐adherent cells. Coating of the immobilization area with mussel adhesive protein was shown to be essential for the process. Intracellular proton and calcium concentrations in different cell classes were simultaneously imaged and the specific activation of T lymphocytes was demonstrated. This method should be especially useful for drug screening due to the small sample volume and high information density.

[1]  J. Waite,et al.  Polyphenolic Substance of Mytilus edulis: Novel Adhesive Containing L-Dopa and Hydroxyproline. , 1981, Science.

[2]  G J Brakenhoff,et al.  Micromanipulation by "multiple" optical traps created by a single fast scanning trap integrated with the bilateral confocal scanning laser microscope. , 1993, Cytometry.

[3]  K. Greulich,et al.  Application of laser optical tweezers in immunology and molecular genetics. , 1991, Cytometry.

[4]  P. Harper,et al.  Fluorescence-based viability assay for studies of reactive drug intermediates. , 1989, Analytical biochemistry.

[5]  J. Broach,et al.  High-throughput screening for drug discovery. , 1996, Nature.

[6]  A. Ashkin,et al.  Optical trapping and manipulation of single cells using infrared laser beams , 1987, Nature.

[7]  J B Shear,et al.  Single cells as biosensors for chemical separations. , 1995, Science.

[8]  S. Chu,et al.  Observation of a single-beam gradient force optical trap for dielectric particles. , 1986, Optics letters.

[9]  R N Zare,et al.  Identification of receptor ligands and receptor subtypes using antagonists in a capillary electrophoresis single-cell biosensor separation system. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[10]  D. Gingell,et al.  Inhibition of cell adhesion by a synthetic polymer adsorbed to glass shown under defined hydrodynamic stress. , 1987, Journal of cell science.

[11]  Stefan Seeger,et al.  Cell patterns for biosensors manufactured with laser tweezers , 1996, European Conference on Biomedical Optics.

[12]  J. Bergh,et al.  Detection of tumor‐specific cytotoxic drug activity IN VITRO using the fluorometric microculture cytotoxicity assay and primary cultures of tumor cells from patients , 1994, International journal of cancer.

[13]  D Kleinfeld,et al.  Controlled outgrowth of dissociated neurons on patterned substrates , 1988, The Journal of neuroscience : the official journal of the Society for Neuroscience.

[14]  Yoh Ogawa,et al.  Optical trapping of small particles using a 1.3-μm compact InGaAsP diode laser , 1991 .

[15]  P. Gardner.  Calcium and T lymphocyte activation , 1989, Cell.

[16]  Ivar Giaever,et al.  A morphological biosensor for mammalian cells , 1993, Nature.

[17]  Daniel I. C. Wang,et al.  Engineering cell shape and function. , 1994, Science.