Abstract

Based on the characteristic of a confocal microscope (CM) that the offset of a pinhole along an optical axis changes the axial intensity response phase, a novel tri-heterodyne confocal microscope is built up by dividing the CM measurement light path into three paths, and using three sets of focusing lenses, detectors and pinholes placed behind, on and before the focal plane to form three detection systems, thereby achieving the axial superresolution imaging and high Signal Noise Ratio (SNR) through pairwise heterodyne subtraction of three intensity signals with given phases received by the three detection systems and data processing. Simulation and experimental results indicate that the new tri-heterodyne confocal microscope reduces the full width at the half maximum of CM axial response curve by more than 50%, results in the significant improvement of CM anti-interference capability, and enables CM to be more suitable for high accuracy bipolar absolute measurement of 3D microstructures and surface contours.

© 2004 Optical Society of America

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References

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  1. T. Wilson, Confocal Microscopy (Academic press limited, 1990).
  2. J. Pawley, Handbook of Biological Confocal Microscopy (Plenun, New York, 1995).
    [Crossref]
  3. Min Gu, Principles of three-dimensional Imaging in Confocal Microscopes (World Scientific Publishing Co.Pte.Ltd., Singapore,1996).
  4. A. Diaspro (ed.) Confocal and Two-Photon Microscopy. Foundations, Applications and Advances (Wiley, New York, 2001).
  5. C.H. Lee and J.P. Wang,“Noninterferometric Differential Confocal Microscopy With 2-nm Depth Resolution,” Opt. Commun. l35,233~237(1997).
    [Crossref]
  6. T. Wilson and S.J. Hewlett, “Superresonlution in confocal scanning microscopy,” Opt.Lett. 16, 1062–1064 (1991).
    [Crossref] [PubMed]
  7. DanielM. de Juana, Jose E. Oti, and Vidal F. Canales, “Design of superresolving continuous phase filters,” Opt. Lett. 28, 607–609 (2003).
    [Crossref] [PubMed]
  8. Liu Li, Deng Xiao-qang, and Wang Gui-ying, “Phase-only optical pupil filter for improving axial resolution in confocal microscopy,” Acta Physica Sinica 50, 48–51(2001).
  9. Tasso R.M. Sales and Michael Morris, “Fundamental limits of optical superresolution,” Opt. Lett. 22, 582–584 (1997).
    [Crossref] [PubMed]
  10. C.J. Schwarz, Y. Kuznetsova, and S.R.J. Brueck, “Imaging interferometric microscopy,” Opt. Lett. 28, 1424–1426 (2003).
    [Crossref] [PubMed]
  11. X. Chen and S.R.J Brueck, “Imaging interferometric lithography: approaching the resolution limits of optics,” Opt.Lett. 24, 124–126 (1999).
    [Crossref]
  12. Vicente Mico, AIDO, and Zeev Zalevsky, “Single-step superresolution by interferometric imaging,” Opt. Express 12, 2589–2596 (2004).http://www.opticsexpress.org/abstract.cfm?URI=OPEX-12-12-2589
    [Crossref] [PubMed]
  13. M. Martinez-Corral, C. Ibáñez-López, G. Saavedra, and Universidad de Valencia, “Axial gain resolutionin optical sectioning fluorescence microscopy by shaded-ring filters,” Opt. Express 15, 1740–1745 (2003). http://www.opticsexpress.org/abstract.cfm?URI=OPEX-11-15-1740
    [Crossref]
  14. Haifeng Wang, “New approach to supperresolution,” Opt. Eng. 40, 851~855(2001).
    [Crossref]
  15. Andrew I. Whiting, Ayman F. Abouraddy, Bahaa E.A. Saleh, and Malvin C. Teich, “Polarization -assisted transverse and axial optical superresolution,” Opt. Express 15, 1714–1723 (2003).
    [Crossref]

2004 (1)

2003 (4)

M. Martinez-Corral, C. Ibáñez-López, G. Saavedra, and Universidad de Valencia, “Axial gain resolutionin optical sectioning fluorescence microscopy by shaded-ring filters,” Opt. Express 15, 1740–1745 (2003). http://www.opticsexpress.org/abstract.cfm?URI=OPEX-11-15-1740
[Crossref]

C.J. Schwarz, Y. Kuznetsova, and S.R.J. Brueck, “Imaging interferometric microscopy,” Opt. Lett. 28, 1424–1426 (2003).
[Crossref] [PubMed]

DanielM. de Juana, Jose E. Oti, and Vidal F. Canales, “Design of superresolving continuous phase filters,” Opt. Lett. 28, 607–609 (2003).
[Crossref] [PubMed]

Andrew I. Whiting, Ayman F. Abouraddy, Bahaa E.A. Saleh, and Malvin C. Teich, “Polarization -assisted transverse and axial optical superresolution,” Opt. Express 15, 1714–1723 (2003).
[Crossref]

2001 (2)

Liu Li, Deng Xiao-qang, and Wang Gui-ying, “Phase-only optical pupil filter for improving axial resolution in confocal microscopy,” Acta Physica Sinica 50, 48–51(2001).

Haifeng Wang, “New approach to supperresolution,” Opt. Eng. 40, 851~855(2001).
[Crossref]

1999 (1)

X. Chen and S.R.J Brueck, “Imaging interferometric lithography: approaching the resolution limits of optics,” Opt.Lett. 24, 124–126 (1999).
[Crossref]

1997 (2)

Tasso R.M. Sales and Michael Morris, “Fundamental limits of optical superresolution,” Opt. Lett. 22, 582–584 (1997).
[Crossref] [PubMed]

C.H. Lee and J.P. Wang,“Noninterferometric Differential Confocal Microscopy With 2-nm Depth Resolution,” Opt. Commun. l35,233~237(1997).
[Crossref]

1991 (1)

T. Wilson and S.J. Hewlett, “Superresonlution in confocal scanning microscopy,” Opt.Lett. 16, 1062–1064 (1991).
[Crossref] [PubMed]

Abouraddy, Ayman F.

Andrew I. Whiting, Ayman F. Abouraddy, Bahaa E.A. Saleh, and Malvin C. Teich, “Polarization -assisted transverse and axial optical superresolution,” Opt. Express 15, 1714–1723 (2003).
[Crossref]

AIDO,

Brueck, S.R.J

X. Chen and S.R.J Brueck, “Imaging interferometric lithography: approaching the resolution limits of optics,” Opt.Lett. 24, 124–126 (1999).
[Crossref]

Brueck, S.R.J.

Canales, Vidal F.

Chen, X.

X. Chen and S.R.J Brueck, “Imaging interferometric lithography: approaching the resolution limits of optics,” Opt.Lett. 24, 124–126 (1999).
[Crossref]

de Juana, DanielM.

de Valencia, Universidad

M. Martinez-Corral, C. Ibáñez-López, G. Saavedra, and Universidad de Valencia, “Axial gain resolutionin optical sectioning fluorescence microscopy by shaded-ring filters,” Opt. Express 15, 1740–1745 (2003). http://www.opticsexpress.org/abstract.cfm?URI=OPEX-11-15-1740
[Crossref]

Gu, Min

Min Gu, Principles of three-dimensional Imaging in Confocal Microscopes (World Scientific Publishing Co.Pte.Ltd., Singapore,1996).

Gui-ying, Wang

Liu Li, Deng Xiao-qang, and Wang Gui-ying, “Phase-only optical pupil filter for improving axial resolution in confocal microscopy,” Acta Physica Sinica 50, 48–51(2001).

Hewlett, S.J.

T. Wilson and S.J. Hewlett, “Superresonlution in confocal scanning microscopy,” Opt.Lett. 16, 1062–1064 (1991).
[Crossref] [PubMed]

Ibáñez-López, C.

M. Martinez-Corral, C. Ibáñez-López, G. Saavedra, and Universidad de Valencia, “Axial gain resolutionin optical sectioning fluorescence microscopy by shaded-ring filters,” Opt. Express 15, 1740–1745 (2003). http://www.opticsexpress.org/abstract.cfm?URI=OPEX-11-15-1740
[Crossref]

Kuznetsova, Y.

Lee, C.H.

C.H. Lee and J.P. Wang,“Noninterferometric Differential Confocal Microscopy With 2-nm Depth Resolution,” Opt. Commun. l35,233~237(1997).
[Crossref]

Li, Liu

Liu Li, Deng Xiao-qang, and Wang Gui-ying, “Phase-only optical pupil filter for improving axial resolution in confocal microscopy,” Acta Physica Sinica 50, 48–51(2001).

Martinez-Corral, M.

M. Martinez-Corral, C. Ibáñez-López, G. Saavedra, and Universidad de Valencia, “Axial gain resolutionin optical sectioning fluorescence microscopy by shaded-ring filters,” Opt. Express 15, 1740–1745 (2003). http://www.opticsexpress.org/abstract.cfm?URI=OPEX-11-15-1740
[Crossref]

Mico, Vicente

Morris, Michael

Oti, Jose E.

Pawley, J.

J. Pawley, Handbook of Biological Confocal Microscopy (Plenun, New York, 1995).
[Crossref]

Saavedra, G.

M. Martinez-Corral, C. Ibáñez-López, G. Saavedra, and Universidad de Valencia, “Axial gain resolutionin optical sectioning fluorescence microscopy by shaded-ring filters,” Opt. Express 15, 1740–1745 (2003). http://www.opticsexpress.org/abstract.cfm?URI=OPEX-11-15-1740
[Crossref]

Saleh, Bahaa E.A.

Andrew I. Whiting, Ayman F. Abouraddy, Bahaa E.A. Saleh, and Malvin C. Teich, “Polarization -assisted transverse and axial optical superresolution,” Opt. Express 15, 1714–1723 (2003).
[Crossref]

Sales, Tasso R.M.

Schwarz, C.J.

Teich, Malvin C.

Andrew I. Whiting, Ayman F. Abouraddy, Bahaa E.A. Saleh, and Malvin C. Teich, “Polarization -assisted transverse and axial optical superresolution,” Opt. Express 15, 1714–1723 (2003).
[Crossref]

Wang, Haifeng

Haifeng Wang, “New approach to supperresolution,” Opt. Eng. 40, 851~855(2001).
[Crossref]

Wang, J.P.

C.H. Lee and J.P. Wang,“Noninterferometric Differential Confocal Microscopy With 2-nm Depth Resolution,” Opt. Commun. l35,233~237(1997).
[Crossref]

Whiting, Andrew I.

Andrew I. Whiting, Ayman F. Abouraddy, Bahaa E.A. Saleh, and Malvin C. Teich, “Polarization -assisted transverse and axial optical superresolution,” Opt. Express 15, 1714–1723 (2003).
[Crossref]

Wilson, T.

T. Wilson and S.J. Hewlett, “Superresonlution in confocal scanning microscopy,” Opt.Lett. 16, 1062–1064 (1991).
[Crossref] [PubMed]

T. Wilson, Confocal Microscopy (Academic press limited, 1990).

Xiao-qang, Deng

Liu Li, Deng Xiao-qang, and Wang Gui-ying, “Phase-only optical pupil filter for improving axial resolution in confocal microscopy,” Acta Physica Sinica 50, 48–51(2001).

Zalevsky, Zeev

Acta Physica Sinica (1)

Liu Li, Deng Xiao-qang, and Wang Gui-ying, “Phase-only optical pupil filter for improving axial resolution in confocal microscopy,” Acta Physica Sinica 50, 48–51(2001).

Opt. Commun. (1)

C.H. Lee and J.P. Wang,“Noninterferometric Differential Confocal Microscopy With 2-nm Depth Resolution,” Opt. Commun. l35,233~237(1997).
[Crossref]

Opt. Eng. (1)

Haifeng Wang, “New approach to supperresolution,” Opt. Eng. 40, 851~855(2001).
[Crossref]

Opt. Express (3)

Andrew I. Whiting, Ayman F. Abouraddy, Bahaa E.A. Saleh, and Malvin C. Teich, “Polarization -assisted transverse and axial optical superresolution,” Opt. Express 15, 1714–1723 (2003).
[Crossref]

Vicente Mico, AIDO, and Zeev Zalevsky, “Single-step superresolution by interferometric imaging,” Opt. Express 12, 2589–2596 (2004).http://www.opticsexpress.org/abstract.cfm?URI=OPEX-12-12-2589
[Crossref] [PubMed]

M. Martinez-Corral, C. Ibáñez-López, G. Saavedra, and Universidad de Valencia, “Axial gain resolutionin optical sectioning fluorescence microscopy by shaded-ring filters,” Opt. Express 15, 1740–1745 (2003). http://www.opticsexpress.org/abstract.cfm?URI=OPEX-11-15-1740
[Crossref]

Opt. Lett. (3)

Opt.Lett. (2)

X. Chen and S.R.J Brueck, “Imaging interferometric lithography: approaching the resolution limits of optics,” Opt.Lett. 24, 124–126 (1999).
[Crossref]

T. Wilson and S.J. Hewlett, “Superresonlution in confocal scanning microscopy,” Opt.Lett. 16, 1062–1064 (1991).
[Crossref] [PubMed]

Other (4)

T. Wilson, Confocal Microscopy (Academic press limited, 1990).

J. Pawley, Handbook of Biological Confocal Microscopy (Plenun, New York, 1995).
[Crossref]

Min Gu, Principles of three-dimensional Imaging in Confocal Microscopes (World Scientific Publishing Co.Pte.Ltd., Singapore,1996).

A. Diaspro (ed.) Confocal and Two-Photon Microscopy. Foundations, Applications and Advances (Wiley, New York, 2001).

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Figures (5)

Fig. 1.
Fig. 1.

Tri-heterodyne CM with axial superresolution.

Fig. 2.
Fig. 2.

Simulated curved surfaces of TCM and CM intensity

Fig. 3.
Fig. 3.

Simulated axial intensity curves with uM =5.21.

Fig. 4.
Fig. 4.

Simulated axial intensity curves when uM =8.0.

Fig. 5.
Fig. 5.

Measured intensity response curves.

Equations (9)

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I 1 v u , + u M = [ 2 0 1 P ( ρ ) · exp ( ju ρ 2 2 ) J 0 ( ρv ) ρdρ ] 2 × [ 2 0 1 P ( ρ ) · exp ( j ρ 2 ( u + u M ) 2 ) J 0 ( ρv ) ρdρ ] 2
u = ( 8 π λ ) z sin 2 ( α 0 2 ) , v ( 2 π λ ) r sin α 0
I 2 v u , u M = [ 2 0 1 P ( ρ ) · exp ( ju ρ 2 2 ) J 0 ( ρv ) ρdρ ] 2 × [ 2 0 1 P ( ρ ) · exp ( j ρ 2 ( u u M ) 2 ) J 0 ( ρv ) ρdρ ] 2
I 3 v u , 0 = [ 2 0 1 P ( ρ ) · exp ( ju ρ 2 2 ) J 0 ( ρv ) ρdρ ] 2 × [ 2 0 1 P ( ρ ) · exp ( ju ρ 2 2 ) J 0 ( ρv ) ρdρ ] 2
{ I A v u = I 3 v u 0 I 2 v u u M I B v u = I 3 v u 0 I 1 v u + u M I C v u = I 2 v u u M I 1 v u + u M
I v u = { I A v u , when I C v u 0 I B v u , when I C v u < 0
k A 0, 0 u M = sin c [ ( u M ) 4 π ] · [ ( u M 4 ) · cos ( u M 4 ) sin ( u M 4 ) ] { ( u M 4 ) } 2
k B 0, 0 u M = sin c [ ( u M ) 4 π ] · [ ( u M 4 ) · cos ( u M 4 ) sin ( u M 4 ) ] { ( u M 4 ) } 2
k C 0, 0 u M = 2 sin c [ ( u M ) 4 π ] · [ ( u M 4 ) · cos ( u M 4 ) sin ( u M 4 ) ] { ( u M 4 ) } 2

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