Elastic Light Scattering from Single Cells: Orientational Dynamics in Optical Trap

Light-scattering diagrams (phase functions) from single living cells and beads suspended in an optical trap were recorded with 30-ms time resolution. The intensity of the scattered light was recorded over an angular range of 0.5–179.5° using an optical setup based on an elliptical mirror and rotatin...

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Bibliographic Details
Published in:Biophysical journal 2004-08, Vol.87 (2), p.1298-1306
Main Authors: Watson, Dakota, Hagen, Norbert, Diver, Jonathan, Marchand, Philippe, Chachisvilis, Mirianas
Format: Article
Language:English
Subjects:
Biochemistry
Cell Polarity - physiology
Cells, Cultured
Cellular biology
Diffusion
Elasticity
Equipment Design
Equipment Failure Analysis
Flow Cytometry - instrumentation
Flow Cytometry - methods
Fluorescence
Granulocytes - cytology
Humans
Leukocytes, Mononuclear - cytology
Melanoma - pathology
Micromanipulation - instrumentation
Micromanipulation - methods
Microscopy, Polarization - instrumentation
Microscopy, Polarization - methods
Optics and Photonics - instrumentation
Phantoms, Imaging
Physical Stimulation - instrumentation
Physical Stimulation - methods
Refractometry - instrumentation
Refractometry - methods
Reproducibility of Results
Scatter diagrams
Scattering, Radiation
Sensitivity and Specificity
Spectroscopy, Imaging, Other Techniques
Spectrum analysis
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Summary:Light-scattering diagrams (phase functions) from single living cells and beads suspended in an optical trap were recorded with 30-ms time resolution. The intensity of the scattered light was recorded over an angular range of 0.5–179.5° using an optical setup based on an elliptical mirror and rotating aperture. Experiments revealed that light-scattering diagrams from biological cells exhibit significant and complex time dependence. We have attributed this dependence to the cell's orientational dynamics within the trap. We have also used experimentally measured phase function information to calculate the time dependence of the optical radiation pressure force on the trapped particle and show how it changes depending on the orientation of the particle. Relevance of these experiments to potential improvement in the sensitivity of label-free flow cytometry is discussed.
ISSN:0006-3495
1542-0086
DOI:10.1529/biophysj.104.042135