Single beam optical trapping integrated in a confocal microscope for biological applications

Koen Visscher, G. J. Brakenhoff

Research output: Contribution to journalArticle

26 Citations (Scopus)

Abstract

Confocal microscopy is very useful in biology because of its three dimensional imaging capacities and has proven to be an excellent tool to study the 3D organization of, for instance, cell structures. This property of confocal microscopy makes it also very suitable for observation during guidance of the three dimensional manipulation of single cells or cell elements. Therefore we decided to integrate a confocal microscope and a single beam optical manipulator into a single instrument. The advantage of optical manipulation over mechanical techniques is that it is non-invasive and therefore may be applied on living (micro-) organisms and cells. The creation of an effective single beam optical trap requires the use of a high numerical aperture (N.A.) objective to focus the laser beam. In this paper we briefly discuss the vertical or axial force exerted on a sphere in a single beam trap. The axial force on a sphere placed on the optical axis, caused by reflection and refraction, is calculated applying a electromagnetic vector diffraction theory to determine the field distribution in the focal region. One of the results is that the particle also experiences a vertical trapping force towards the focusing lens when it is in the strongly convergent part of the field in addition to the known negative signed trapping force in the divergent part of the field. Further we describe an instrumental approach to realize optical trapping in which the optical trap position is controlled by moving the focusing objective only. At present we have demonstrated the practical use of the presented optical trap control on the spatial x, y, z manipulation of micro-sized polystyrene spheres (1-7 μm) and of yeast cells.

Original languageEnglish (US)
Pages (from-to)486-491
Number of pages6
JournalCytometry
Volume12
Issue number6
StatePublished - 1991
Externally publishedYes

Fingerprint

Optical Tweezers
Confocal Microscopy
Three-Dimensional Imaging
Electromagnetic Phenomena
Polystyrenes
Lenses
Lasers
Yeasts
Observation

Keywords

  • confocal microscopy
  • electromagnetic diffraction
  • geometrical optics
  • high numerical aperture
  • non-invasive manipulation
  • optical trapping

ASJC Scopus subject areas

  • Hematology
  • Cell Biology
  • Pathology and Forensic Medicine
  • Biophysics
  • Endocrinology

Cite this

Single beam optical trapping integrated in a confocal microscope for biological applications. / Visscher, Koen; Brakenhoff, G. J.

In: Cytometry, Vol. 12, No. 6, 1991, p. 486-491.

Research output: Contribution to journalArticle

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