Abstract

Following the recently reported trapping of biological particles by finely focused laser beams, we report on the automated micromanipulation of cells and other microscopic particles by purely optical means as well as on a newly observed interaction between particles in the trapping beam. A simple instrument is described which allows single cells to be positioned with high accuracy, transported over several millimeters, and automatically sorted on the basis of their optical properties. These operations are performed inside a small enclosed chamber without mechanical contact or significant fluid flow. Potential applications of this technique in experimental cell biology are discussed.

© 1987 Optical Society of America

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References

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  1. J. A. Steinkamp, “Flow Cytometry,” Rev. Sci. Instrum. 55, 1375 (1984).
    [Crossref]
  2. A. Ashkin, “Acceleration and Trapping of Particles by Radiation Pressure,” Phys. Rev. Lett. 24, 156 (1970).
    [Crossref]
  3. A. Ashkin, J. M. Dziedzic, “Optical Trapping and Manipulation of Viruses and Bacteria,” Science 235, 1517 (1987).
    [Crossref] [PubMed]
  4. A. Ashkin, “Applications of Laser Radiation Pressure,” Science 210, 1081 (1980).
    [Crossref] [PubMed]
  5. A. Ashkin, J. M. Dziedzic, “Stability of Optical Levitation by Radiation Pressure,” Appl. Phys. Lett. 24, 586 (1974).
    [Crossref]
  6. A. Ashkin, J. M. Dziedzic, “Optical Levitation by Radiation Pressure,” Appl. Phys. Lett. 19, 283 (1971).
    [Crossref]
  7. H. Kogelnik, T. Li, “Laser Beams and Resonators,” Appl. Opt. 5, 1550 (1966).
    [Crossref] [PubMed]
  8. H. A. Crissman, J. A. Steinkamp, “Rapid, One Step Staining Procedures for Analysis of Cellular DNA and Protein by Single and Dual Laser Flow Cytometry,” Cytometry 3, 84 (1982).
    [Crossref] [PubMed]
  9. A. Ashkin, J. M. Dziedzic, “Optical Levitation of Liquid Drops by Radiation Pressure,” Science 187, 1073 (1975).
    [Crossref] [PubMed]
  10. A. Ashkin, J. M. Dziedzic, “Observation of Light Scattering from Nonspherical Particles using Optical Levitation,” Appl. Opt. 19, 660 (1980).
    [Crossref] [PubMed]
  11. G. Roosen, “La levitation optique de spheres,” Can. J. Phys. 57, 1260 (1979).
    [Crossref]
  12. A. Brunsting, P. F. Mullaney, “Differential Light Scattering from Spherical Mammalian Cells,” Biophys. J. 14, 439 (1974).
    [Crossref] [PubMed]
  13. K. W. Keohane, W. K. Metcalf, “The Cytoplasmic Refractive Index of Lymphocytes, its Significance and its Changes during Active Immunization,” Q. J. Exp. Physiol. 44, 343 (1959).

1987 (1)

A. Ashkin, J. M. Dziedzic, “Optical Trapping and Manipulation of Viruses and Bacteria,” Science 235, 1517 (1987).
[Crossref] [PubMed]

1984 (1)

J. A. Steinkamp, “Flow Cytometry,” Rev. Sci. Instrum. 55, 1375 (1984).
[Crossref]

1982 (1)

H. A. Crissman, J. A. Steinkamp, “Rapid, One Step Staining Procedures for Analysis of Cellular DNA and Protein by Single and Dual Laser Flow Cytometry,” Cytometry 3, 84 (1982).
[Crossref] [PubMed]

1980 (2)

1979 (1)

G. Roosen, “La levitation optique de spheres,” Can. J. Phys. 57, 1260 (1979).
[Crossref]

1975 (1)

A. Ashkin, J. M. Dziedzic, “Optical Levitation of Liquid Drops by Radiation Pressure,” Science 187, 1073 (1975).
[Crossref] [PubMed]

1974 (2)

A. Ashkin, J. M. Dziedzic, “Stability of Optical Levitation by Radiation Pressure,” Appl. Phys. Lett. 24, 586 (1974).
[Crossref]

A. Brunsting, P. F. Mullaney, “Differential Light Scattering from Spherical Mammalian Cells,” Biophys. J. 14, 439 (1974).
[Crossref] [PubMed]

1971 (1)

A. Ashkin, J. M. Dziedzic, “Optical Levitation by Radiation Pressure,” Appl. Phys. Lett. 19, 283 (1971).
[Crossref]

1970 (1)

A. Ashkin, “Acceleration and Trapping of Particles by Radiation Pressure,” Phys. Rev. Lett. 24, 156 (1970).
[Crossref]

1966 (1)

1959 (1)

K. W. Keohane, W. K. Metcalf, “The Cytoplasmic Refractive Index of Lymphocytes, its Significance and its Changes during Active Immunization,” Q. J. Exp. Physiol. 44, 343 (1959).

Ashkin, A.

A. Ashkin, J. M. Dziedzic, “Optical Trapping and Manipulation of Viruses and Bacteria,” Science 235, 1517 (1987).
[Crossref] [PubMed]

A. Ashkin, “Applications of Laser Radiation Pressure,” Science 210, 1081 (1980).
[Crossref] [PubMed]

A. Ashkin, J. M. Dziedzic, “Observation of Light Scattering from Nonspherical Particles using Optical Levitation,” Appl. Opt. 19, 660 (1980).
[Crossref] [PubMed]

A. Ashkin, J. M. Dziedzic, “Optical Levitation of Liquid Drops by Radiation Pressure,” Science 187, 1073 (1975).
[Crossref] [PubMed]

A. Ashkin, J. M. Dziedzic, “Stability of Optical Levitation by Radiation Pressure,” Appl. Phys. Lett. 24, 586 (1974).
[Crossref]

A. Ashkin, J. M. Dziedzic, “Optical Levitation by Radiation Pressure,” Appl. Phys. Lett. 19, 283 (1971).
[Crossref]

A. Ashkin, “Acceleration and Trapping of Particles by Radiation Pressure,” Phys. Rev. Lett. 24, 156 (1970).
[Crossref]

Brunsting, A.

A. Brunsting, P. F. Mullaney, “Differential Light Scattering from Spherical Mammalian Cells,” Biophys. J. 14, 439 (1974).
[Crossref] [PubMed]

Crissman, H. A.

H. A. Crissman, J. A. Steinkamp, “Rapid, One Step Staining Procedures for Analysis of Cellular DNA and Protein by Single and Dual Laser Flow Cytometry,” Cytometry 3, 84 (1982).
[Crossref] [PubMed]

Dziedzic, J. M.

A. Ashkin, J. M. Dziedzic, “Optical Trapping and Manipulation of Viruses and Bacteria,” Science 235, 1517 (1987).
[Crossref] [PubMed]

A. Ashkin, J. M. Dziedzic, “Observation of Light Scattering from Nonspherical Particles using Optical Levitation,” Appl. Opt. 19, 660 (1980).
[Crossref] [PubMed]

A. Ashkin, J. M. Dziedzic, “Optical Levitation of Liquid Drops by Radiation Pressure,” Science 187, 1073 (1975).
[Crossref] [PubMed]

A. Ashkin, J. M. Dziedzic, “Stability of Optical Levitation by Radiation Pressure,” Appl. Phys. Lett. 24, 586 (1974).
[Crossref]

A. Ashkin, J. M. Dziedzic, “Optical Levitation by Radiation Pressure,” Appl. Phys. Lett. 19, 283 (1971).
[Crossref]

Keohane, K. W.

K. W. Keohane, W. K. Metcalf, “The Cytoplasmic Refractive Index of Lymphocytes, its Significance and its Changes during Active Immunization,” Q. J. Exp. Physiol. 44, 343 (1959).

Kogelnik, H.

Li, T.

Metcalf, W. K.

K. W. Keohane, W. K. Metcalf, “The Cytoplasmic Refractive Index of Lymphocytes, its Significance and its Changes during Active Immunization,” Q. J. Exp. Physiol. 44, 343 (1959).

Mullaney, P. F.

A. Brunsting, P. F. Mullaney, “Differential Light Scattering from Spherical Mammalian Cells,” Biophys. J. 14, 439 (1974).
[Crossref] [PubMed]

Roosen, G.

G. Roosen, “La levitation optique de spheres,” Can. J. Phys. 57, 1260 (1979).
[Crossref]

Steinkamp, J. A.

J. A. Steinkamp, “Flow Cytometry,” Rev. Sci. Instrum. 55, 1375 (1984).
[Crossref]

H. A. Crissman, J. A. Steinkamp, “Rapid, One Step Staining Procedures for Analysis of Cellular DNA and Protein by Single and Dual Laser Flow Cytometry,” Cytometry 3, 84 (1982).
[Crossref] [PubMed]

Appl. Opt. (2)

Appl. Phys. Lett. (2)

A. Ashkin, J. M. Dziedzic, “Stability of Optical Levitation by Radiation Pressure,” Appl. Phys. Lett. 24, 586 (1974).
[Crossref]

A. Ashkin, J. M. Dziedzic, “Optical Levitation by Radiation Pressure,” Appl. Phys. Lett. 19, 283 (1971).
[Crossref]

Biophys. J. (1)

A. Brunsting, P. F. Mullaney, “Differential Light Scattering from Spherical Mammalian Cells,” Biophys. J. 14, 439 (1974).
[Crossref] [PubMed]

Can. J. Phys. (1)

G. Roosen, “La levitation optique de spheres,” Can. J. Phys. 57, 1260 (1979).
[Crossref]

Cytometry (1)

H. A. Crissman, J. A. Steinkamp, “Rapid, One Step Staining Procedures for Analysis of Cellular DNA and Protein by Single and Dual Laser Flow Cytometry,” Cytometry 3, 84 (1982).
[Crossref] [PubMed]

Phys. Rev. Lett. (1)

A. Ashkin, “Acceleration and Trapping of Particles by Radiation Pressure,” Phys. Rev. Lett. 24, 156 (1970).
[Crossref]

Q. J. Exp. Physiol. (1)

K. W. Keohane, W. K. Metcalf, “The Cytoplasmic Refractive Index of Lymphocytes, its Significance and its Changes during Active Immunization,” Q. J. Exp. Physiol. 44, 343 (1959).

Rev. Sci. Instrum. (1)

J. A. Steinkamp, “Flow Cytometry,” Rev. Sci. Instrum. 55, 1375 (1984).
[Crossref]

Science (3)

A. Ashkin, J. M. Dziedzic, “Optical Trapping and Manipulation of Viruses and Bacteria,” Science 235, 1517 (1987).
[Crossref] [PubMed]

A. Ashkin, “Applications of Laser Radiation Pressure,” Science 210, 1081 (1980).
[Crossref] [PubMed]

A. Ashkin, J. M. Dziedzic, “Optical Levitation of Liquid Drops by Radiation Pressure,” Science 187, 1073 (1975).
[Crossref] [PubMed]

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Figures (6)

Fig. 1
Fig. 1

Simplified diagram of the automated optical manipulator: S, beam splitter; M, mirrors; L1, L2, lenses; C, chamber; A, aperture; AO, acoustooptic modulator; SP, short-pass filter; LP, long-pass filter; MS, microscope; VC, video camera; OF, optical fiber; PMT, photomultiplier; AMP, amplifier; PSA, pulse shape analyzer; AOD, acoustooptic modulator driver. Although they coincide in the drawing, the paths of the deflection and probe beams differ slightly to achieve a separation of ∼100 μm as they intersect the propulsion beam inside the chamber. The propulsion and deflection beams have a wavelength of 488 nm, while the wavelength of the probe beam is 633 nm. Focusing lenses L1 and L2 have a focal length of 50 mm.

Fig. 2
Fig. 2

Top view of the chamber (a) and horizontal cross section at the level of the sample injection port (b). The sample injection port is denoted by SP. The entrance and exit windows can be seen on the four sides of the chamber. The circles are elution and rinsing ports. The inner square in (a) denotes the top window through which microscopic observations are made. The channels seen in (a) are machined into the surface of the chamber and are 3.5 mm deep and 1 mm wide. The scale bar is 10 mm long. The chamber is made out of brass, and the windows are cut from 170-μm thick coverslips. The windows are glued to the chamber with an epoxy adhesive.

Fig. 3
Fig. 3

Deflection of a single 7.5-μm polystyrene microsphere: (a) The microsphere carried by the propulsion beam crosses the probe beam. One can see the diffraction fringes projected onto the bottom of one of the chamber channels. (b) The microsphere has just been deflected. The deflection beam, which is still at high intensity, can be easily seen. (c) The deflected particle is being transported upward by the deflection beam. The beam is now at its lower intensity level. The position of the particle is marked in (b) and (c) by an arrow.

Fig. 4
Fig. 4

Rejection of a 7.5-μm polystyrene microsphere doublet. (a) The doublet, with its axis parallel to the propulsion beam, crosses the probe beam. The doublet intereference fringes can be easily seen. (b) The doublet has traveled past the deflection beam, which has been kept at the lower intensity level to prevent deflection. The position of the doublet is marked by an arrow. Two additional particles can be seen in the propulsion beam to the left of the doublet.

Fig. 5
Fig. 5

Deflection of an ethanol-fixed CHO cell. (a) The cell crosses the probe beam. (b) The cell, marked by an arrow, has just been deflected as a consequence of the amplitude of its 90° light scatter pulse falling within the discriminator window. The cells were stained with 10-ng/mliter FITC to improve their visibility.

Fig. 6
Fig. 6

Beam-mediated interaction between two CHO cells. (a) The cell to the left has just entered the propulsion beam. (b) The separation between the cells has reached its minimum value. (c) The separation in (b) is being maintained while the two cells travel in the propulsion beam. The position of the two cells is marked by a horizontal bar.

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