Abstract

We report on the development and testing of a compact laser tweezers Raman spectroscopy (LTRS) system. The system combines optical trapping and near-infrared Raman spectroscopy for manipulation and identification of single biological cells in solution. A low-power diode laser at 785 nm was used for both trapping and excitation for Raman spectroscopy of the suspended microscopic particles. The design of the LTRS system provides high sensitivity and permits real-time spectroscopic measurements of the biological sample. The system was calibrated by use of polystyrene microbeads and tested on living blood cells and on both living and dead yeast cells. As expected, different images and Raman spectra were observed for the different cells. The LTRS system may provide a valuable tool for the study of fundamental cellular processes and the diagnosis of cellular disorders.

© 2002 Optical Society of America

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2001 (2)

K. Ajito and K. Torimitsu, Trends Anal. Chem. 20, 255 (2001).
[CrossRef]

B. R. Wood, B. Tait, and D. McNaughton, Biochim. Biophys. Acta 1539, 58 (2001).
[CrossRef] [PubMed]

2000 (1)

J. Guck, R. Ananthakrishnan, T. J. Moon, C. C. Cunningham, and J. Kas, Phys. Rev. Lett. 84, 5451 (2000).
[CrossRef] [PubMed]

1999 (3)

K. Visscher, M. J. Schnitzer, and S. M. Block, Nature 400, 184 (1999).
[CrossRef] [PubMed]

K. Neuman, E. H. Chadd, G. F. Liou, K. Bergman, and S. M. Block, Biophys. J. 77, 2656 (1999).

N. Kikatani, H.-B. Kim, S. Habuchi, and M. Chiba, Trends Anal. Chem. 18, 675 (1999).
[CrossRef]

1998 (1)

1996 (2)

Y. Q. Li, W. H. Burkett, and M. Xiao, Opt. Lett. 21, 982 (1996).
[CrossRef] [PubMed]

S. R. Hawi, W. B. Campbell, A. Kajdacsy-Balla, R. Murphy, F. Adar, and K. Nithipatikom, Cancer Lett. 110, 35 (1996).
[CrossRef] [PubMed]

1995 (2)

K. Konig, H. Liang, M. W. Berns, and B. J. Tromberg, Nature 377, 20 (1995).
[CrossRef]

Y. Liu, G. J. Sonek, M. W. Berns, K. Konig, and B. J. Tromberg, Opt. Lett. 20, 2246 (1995).
[CrossRef]

1994 (2)

M. Lankers, J. Popp, and W. Kiefer, Appl. Spectrosc. 48, 1166 (1994).
[CrossRef]

K. Svoboda and S. M. Block, Annu. Rev. Biophys. Biomol. Struct. 23, 247 (1994).
[CrossRef]

1987 (1)

A. Ashkin, K. M. Dziedzic, and T. Yamane, Nature 330, 769 (1987).
[CrossRef] [PubMed]

1986 (1)

Adar, F.

S. R. Hawi, W. B. Campbell, A. Kajdacsy-Balla, R. Murphy, F. Adar, and K. Nithipatikom, Cancer Lett. 110, 35 (1996).
[CrossRef] [PubMed]

Ajito, K.

K. Ajito and K. Torimitsu, Trends Anal. Chem. 20, 255 (2001).
[CrossRef]

K. Ajito, Appl. Spectrosc. 52, 339 (1998).
[CrossRef]

Ananthakrishnan, R.

J. Guck, R. Ananthakrishnan, T. J. Moon, C. C. Cunningham, and J. Kas, Phys. Rev. Lett. 84, 5451 (2000).
[CrossRef] [PubMed]

Ashkin, A.

Bergman, K.

K. Neuman, E. H. Chadd, G. F. Liou, K. Bergman, and S. M. Block, Biophys. J. 77, 2656 (1999).

Berns, M. W.

K. Konig, H. Liang, M. W. Berns, and B. J. Tromberg, Nature 377, 20 (1995).
[CrossRef]

Y. Liu, G. J. Sonek, M. W. Berns, K. Konig, and B. J. Tromberg, Opt. Lett. 20, 2246 (1995).
[CrossRef]

Bjorkholm, J. E.

Block, S. M.

K. Visscher, M. J. Schnitzer, and S. M. Block, Nature 400, 184 (1999).
[CrossRef] [PubMed]

K. Neuman, E. H. Chadd, G. F. Liou, K. Bergman, and S. M. Block, Biophys. J. 77, 2656 (1999).

K. Svoboda and S. M. Block, Annu. Rev. Biophys. Biomol. Struct. 23, 247 (1994).
[CrossRef]

Burkett, W. H.

Campbell, W. B.

S. R. Hawi, W. B. Campbell, A. Kajdacsy-Balla, R. Murphy, F. Adar, and K. Nithipatikom, Cancer Lett. 110, 35 (1996).
[CrossRef] [PubMed]

Chadd, E. H.

K. Neuman, E. H. Chadd, G. F. Liou, K. Bergman, and S. M. Block, Biophys. J. 77, 2656 (1999).

Chiba, M.

N. Kikatani, H.-B. Kim, S. Habuchi, and M. Chiba, Trends Anal. Chem. 18, 675 (1999).
[CrossRef]

Chu, S.

Cunningham, C. C.

J. Guck, R. Ananthakrishnan, T. J. Moon, C. C. Cunningham, and J. Kas, Phys. Rev. Lett. 84, 5451 (2000).
[CrossRef] [PubMed]

Dziedzic, J. M.

Dziedzic, K. M.

A. Ashkin, K. M. Dziedzic, and T. Yamane, Nature 330, 769 (1987).
[CrossRef] [PubMed]

Guck, J.

J. Guck, R. Ananthakrishnan, T. J. Moon, C. C. Cunningham, and J. Kas, Phys. Rev. Lett. 84, 5451 (2000).
[CrossRef] [PubMed]

Habuchi, S.

N. Kikatani, H.-B. Kim, S. Habuchi, and M. Chiba, Trends Anal. Chem. 18, 675 (1999).
[CrossRef]

Hawi, S. R.

S. R. Hawi, W. B. Campbell, A. Kajdacsy-Balla, R. Murphy, F. Adar, and K. Nithipatikom, Cancer Lett. 110, 35 (1996).
[CrossRef] [PubMed]

Kajdacsy-Balla, A.

S. R. Hawi, W. B. Campbell, A. Kajdacsy-Balla, R. Murphy, F. Adar, and K. Nithipatikom, Cancer Lett. 110, 35 (1996).
[CrossRef] [PubMed]

Kas, J.

J. Guck, R. Ananthakrishnan, T. J. Moon, C. C. Cunningham, and J. Kas, Phys. Rev. Lett. 84, 5451 (2000).
[CrossRef] [PubMed]

Kiefer, W.

Kikatani, N.

N. Kikatani, H.-B. Kim, S. Habuchi, and M. Chiba, Trends Anal. Chem. 18, 675 (1999).
[CrossRef]

Kim, H.-B.

N. Kikatani, H.-B. Kim, S. Habuchi, and M. Chiba, Trends Anal. Chem. 18, 675 (1999).
[CrossRef]

Konig, K.

K. Konig, H. Liang, M. W. Berns, and B. J. Tromberg, Nature 377, 20 (1995).
[CrossRef]

Y. Liu, G. J. Sonek, M. W. Berns, K. Konig, and B. J. Tromberg, Opt. Lett. 20, 2246 (1995).
[CrossRef]

Lankers, M.

Li, Y. Q.

Liang, H.

K. Konig, H. Liang, M. W. Berns, and B. J. Tromberg, Nature 377, 20 (1995).
[CrossRef]

Liou, G. F.

K. Neuman, E. H. Chadd, G. F. Liou, K. Bergman, and S. M. Block, Biophys. J. 77, 2656 (1999).

Liu, Y.

McNaughton, D.

B. R. Wood, B. Tait, and D. McNaughton, Biochim. Biophys. Acta 1539, 58 (2001).
[CrossRef] [PubMed]

Moon, T. J.

J. Guck, R. Ananthakrishnan, T. J. Moon, C. C. Cunningham, and J. Kas, Phys. Rev. Lett. 84, 5451 (2000).
[CrossRef] [PubMed]

Murphy, R.

S. R. Hawi, W. B. Campbell, A. Kajdacsy-Balla, R. Murphy, F. Adar, and K. Nithipatikom, Cancer Lett. 110, 35 (1996).
[CrossRef] [PubMed]

Neuman, K.

K. Neuman, E. H. Chadd, G. F. Liou, K. Bergman, and S. M. Block, Biophys. J. 77, 2656 (1999).

Nithipatikom, K.

S. R. Hawi, W. B. Campbell, A. Kajdacsy-Balla, R. Murphy, F. Adar, and K. Nithipatikom, Cancer Lett. 110, 35 (1996).
[CrossRef] [PubMed]

Popp, J.

Schnitzer, M. J.

K. Visscher, M. J. Schnitzer, and S. M. Block, Nature 400, 184 (1999).
[CrossRef] [PubMed]

Sonek, G. J.

Svoboda, K.

K. Svoboda and S. M. Block, Annu. Rev. Biophys. Biomol. Struct. 23, 247 (1994).
[CrossRef]

Tait, B.

B. R. Wood, B. Tait, and D. McNaughton, Biochim. Biophys. Acta 1539, 58 (2001).
[CrossRef] [PubMed]

Torimitsu, K.

K. Ajito and K. Torimitsu, Trends Anal. Chem. 20, 255 (2001).
[CrossRef]

Tromberg, B. J.

Y. Liu, G. J. Sonek, M. W. Berns, K. Konig, and B. J. Tromberg, Opt. Lett. 20, 2246 (1995).
[CrossRef]

K. Konig, H. Liang, M. W. Berns, and B. J. Tromberg, Nature 377, 20 (1995).
[CrossRef]

Visscher, K.

K. Visscher, M. J. Schnitzer, and S. M. Block, Nature 400, 184 (1999).
[CrossRef] [PubMed]

Wood, B. R.

B. R. Wood, B. Tait, and D. McNaughton, Biochim. Biophys. Acta 1539, 58 (2001).
[CrossRef] [PubMed]

Xiao, M.

Yamane, T.

A. Ashkin, K. M. Dziedzic, and T. Yamane, Nature 330, 769 (1987).
[CrossRef] [PubMed]

Annu. Rev. Biophys. Biomol. Struct. (1)

K. Svoboda and S. M. Block, Annu. Rev. Biophys. Biomol. Struct. 23, 247 (1994).
[CrossRef]

Appl. Spectrosc. (2)

Biochim. Biophys. Acta (1)

B. R. Wood, B. Tait, and D. McNaughton, Biochim. Biophys. Acta 1539, 58 (2001).
[CrossRef] [PubMed]

Biophys. J. (1)

K. Neuman, E. H. Chadd, G. F. Liou, K. Bergman, and S. M. Block, Biophys. J. 77, 2656 (1999).

Cancer Lett. (1)

S. R. Hawi, W. B. Campbell, A. Kajdacsy-Balla, R. Murphy, F. Adar, and K. Nithipatikom, Cancer Lett. 110, 35 (1996).
[CrossRef] [PubMed]

Nature (3)

K. Konig, H. Liang, M. W. Berns, and B. J. Tromberg, Nature 377, 20 (1995).
[CrossRef]

K. Visscher, M. J. Schnitzer, and S. M. Block, Nature 400, 184 (1999).
[CrossRef] [PubMed]

A. Ashkin, K. M. Dziedzic, and T. Yamane, Nature 330, 769 (1987).
[CrossRef] [PubMed]

Opt. Lett. (3)

Phys. Rev. Lett. (1)

J. Guck, R. Ananthakrishnan, T. J. Moon, C. C. Cunningham, and J. Kas, Phys. Rev. Lett. 84, 5451 (2000).
[CrossRef] [PubMed]

Trends Anal. Chem. (2)

N. Kikatani, H.-B. Kim, S. Habuchi, and M. Chiba, Trends Anal. Chem. 18, 675 (1999).
[CrossRef]

K. Ajito and K. Torimitsu, Trends Anal. Chem. 20, 255 (2001).
[CrossRef]

Other (1)

M. P. Sheetz, ed., Laser Tweezers in Cell Biology, Vol. 55 of Methods in Cell Biology (Academic, San Diego, Calif., 1998).

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Figures (4)

Fig. 1
Fig. 1

Experimental setup: DL, diode laser; IF, interference filter; DM, dichroic mirror; HNF1, HNF2, holographic notch filters; M’s, reflection mirrors; L, lens; BS, beam splitter; Obj, objective lens; EP, eyepiece; VC, video camera; Mod, modulation current pulses; Xe, green-filtered xenon illumination light.

Fig. 2
Fig. 2

Raman spectrum and (inset) image of a single polystyrene latex bead of 2.03µm diameter in an optical trap. The trapping power is 2.0 mW and the CCD acquisition time is 2.0 s with 20-mW excitation.

Fig. 3
Fig. 3

NIR Raman spectra and image of a single RBC (erythrocyte) in saline solution. The acquisition time is 5.0 s with 20-mW excitation. Curve (a) is the spectrum recorded when a RBC is trapped, curve (b) is the background without a RBC in the trap, and curve (c) is the subtraction between (a) and (b), magnified by 2 for display.

Fig. 4
Fig. 4

NIR Raman spectra and images of a single living yeast cell and a dead yeast cell in solution. A significant difference can be seen in the Raman spectra of the living and the dead cells.

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