Abstract

A diffraction analysis is presented for image formation in confocal microscopy using the divided aperture technique, which uses two D-shaped apertures (also called specular microscopy). The effects of increasing the width of a divider, that separates the two D shapes, are investigated. As the width is increased, the resolution degrades. The efficiency of singly-scattered light rejection is not improved with increased width.

© 2008 Optical Society of America

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    [CrossRef]
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2007

2006

2004

J. Huisken, J. Swoger, F. Del Bene, J. Wittbrodt, and E. H. K. Stelzer, "Optical sectioning deep inside live embryos by selective plane illumination microscopy," Science 305, 1007-1009 (2004).
[CrossRef] [PubMed]

2003

2001

C. J. R. Sheppard and M. D. Sharma, "Integrated intensity, and imaging through scattering media," J. Mod. Opt. 48, 1517-1525 (2001).

2000

1996

1995

E. H. K. Stelzer, S. Lindek, S. Albrecht, R. Pick, G. Ritter, N. J. Salmon, and R. Stricker, "A new tool for the observation of embryos and other large specimens - confocal theta-fluorescence microscopy," J. Microsc. 179, 1-10 (1995).
[CrossRef]

1993

A. H. Voie, D. H. Burns, and F. A. Spelman, "Orthogonal-plane fluorescence optical sectioning: three-dimensional imaging of macroscopic biological specimens," J. Microsc. 170, 229-236 (1993).
[CrossRef] [PubMed]

M. Gu, C. J. R. Sheppard, and H. Zhou, "Optimization of axial resolution in confocal imaging using annular pupils," Optik 93, 87-90 (1993).

X. S. Gan and C. J. R. Sheppard, "Detectability: A new criterion for evaluation of the confocal microscope," Scanning 15, 187-192 (1993).
[CrossRef]

1980

1978

1974

D. M. Maurice, "A scanning slit optical microscope," Invest. Ophthalmol. 13, 1033-1037 (1974).
[PubMed]

1968

D. M. Maurice, "Cellular membrane activity in the corneal endothelium of the intact eye," Experientia 15, 1094-1095 (1968).
[CrossRef]

1953

E. H. Linfoot and E. Wolf, "Diffraction images in systems with an annular aperture," Proc. Phys. Soc. B 66, 145-149 (1953).
[CrossRef]

1940

H. Goldman, "Spaltlampenphotographie und -photometrie," Ophthalmologica 98, 257-270 (1940).
[CrossRef]

1903

H. Siedentopf and R. Zsigmondy, "Uber Sichtbarmachung und Grössenbestimmung ultramikroskopischer Teilchen, mit besonderer Anwendung auf Goldrubingläser," Annalen der Physik 10, 1-39 (1903).

Aguilar, J. F.

Albrecht, S.

E. H. K. Stelzer, S. Lindek, S. Albrecht, R. Pick, G. Ritter, N. J. Salmon, and R. Stricker, "A new tool for the observation of embryos and other large specimens - confocal theta-fluorescence microscopy," J. Microsc. 179, 1-10 (1995).
[CrossRef]

Burns, D. H.

A. H. Voie, D. H. Burns, and F. A. Spelman, "Orthogonal-plane fluorescence optical sectioning: three-dimensional imaging of macroscopic biological specimens," J. Microsc. 170, 229-236 (1993).
[CrossRef] [PubMed]

Contag, C. H.

Del Bene, F.

J. Huisken, J. Swoger, F. Del Bene, J. Wittbrodt, and E. H. K. Stelzer, "Optical sectioning deep inside live embryos by selective plane illumination microscopy," Science 305, 1007-1009 (2004).
[CrossRef] [PubMed]

DiMarzio, C. A.

Dwyer, P. J.

Gan, X. S.

X. S. Gan and C. J. R. Sheppard, "Detectability: A new criterion for evaluation of the confocal microscope," Scanning 15, 187-192 (1993).
[CrossRef]

Goldman, H.

H. Goldman, "Spaltlampenphotographie und -photometrie," Ophthalmologica 98, 257-270 (1940).
[CrossRef]

Gu, M.

M. Gu, T. Tannous, and C. J. R. Sheppard, "Effect of numerical aperture, pinhole size and annular pupil on confocal imaging through highly scattering media," Opt. Lett. 21, 312-314 (1996).
[CrossRef] [PubMed]

M. Gu, C. J. R. Sheppard, and H. Zhou, "Optimization of axial resolution in confocal imaging using annular pupils," Optik 93, 87-90 (1993).

Huisken, J.

J. Huisken, J. Swoger, F. Del Bene, J. Wittbrodt, and E. H. K. Stelzer, "Optical sectioning deep inside live embryos by selective plane illumination microscopy," Science 305, 1007-1009 (2004).
[CrossRef] [PubMed]

Kino, G. S.

Koester, C. J.

Laczik, Z.

Lera, M.

Lindek, S.

E. H. K. Stelzer, S. Lindek, S. Albrecht, R. Pick, G. Ritter, N. J. Salmon, and R. Stricker, "A new tool for the observation of embryos and other large specimens - confocal theta-fluorescence microscopy," J. Microsc. 179, 1-10 (1995).
[CrossRef]

Linfoot, E. H.

E. H. Linfoot and E. Wolf, "Diffraction images in systems with an annular aperture," Proc. Phys. Soc. B 66, 145-149 (1953).
[CrossRef]

Mandella, M. J.

Maurice, D. M.

D. M. Maurice, "A scanning slit optical microscope," Invest. Ophthalmol. 13, 1033-1037 (1974).
[PubMed]

D. M. Maurice, "Cellular membrane activity in the corneal endothelium of the intact eye," Experientia 15, 1094-1095 (1968).
[CrossRef]

Pick, R.

E. H. K. Stelzer, S. Lindek, S. Albrecht, R. Pick, G. Ritter, N. J. Salmon, and R. Stricker, "A new tool for the observation of embryos and other large specimens - confocal theta-fluorescence microscopy," J. Microsc. 179, 1-10 (1995).
[CrossRef]

Rajadhyaksha, M.

Ritter, G.

E. H. K. Stelzer, S. Lindek, S. Albrecht, R. Pick, G. Ritter, N. J. Salmon, and R. Stricker, "A new tool for the observation of embryos and other large specimens - confocal theta-fluorescence microscopy," J. Microsc. 179, 1-10 (1995).
[CrossRef]

Salmon, N. J.

E. H. K. Stelzer, S. Lindek, S. Albrecht, R. Pick, G. Ritter, N. J. Salmon, and R. Stricker, "A new tool for the observation of embryos and other large specimens - confocal theta-fluorescence microscopy," J. Microsc. 179, 1-10 (1995).
[CrossRef]

Sharma, M. D.

C. J. R. Sheppard and M. D. Sharma, "Integrated intensity, and imaging through scattering media," J. Mod. Opt. 48, 1517-1525 (2001).

Sheppard, C. J. R.

Siedentopf, H.

H. Siedentopf and R. Zsigmondy, "Uber Sichtbarmachung und Grössenbestimmung ultramikroskopischer Teilchen, mit besonderer Anwendung auf Goldrubingläser," Annalen der Physik 10, 1-39 (1903).

Spelman, F. A.

A. H. Voie, D. H. Burns, and F. A. Spelman, "Orthogonal-plane fluorescence optical sectioning: three-dimensional imaging of macroscopic biological specimens," J. Microsc. 170, 229-236 (1993).
[CrossRef] [PubMed]

Stelzer, E. H. K.

J. Huisken, J. Swoger, F. Del Bene, J. Wittbrodt, and E. H. K. Stelzer, "Optical sectioning deep inside live embryos by selective plane illumination microscopy," Science 305, 1007-1009 (2004).
[CrossRef] [PubMed]

E. H. K. Stelzer, S. Lindek, S. Albrecht, R. Pick, G. Ritter, N. J. Salmon, and R. Stricker, "A new tool for the observation of embryos and other large specimens - confocal theta-fluorescence microscopy," J. Microsc. 179, 1-10 (1995).
[CrossRef]

Stricker, R.

E. H. K. Stelzer, S. Lindek, S. Albrecht, R. Pick, G. Ritter, N. J. Salmon, and R. Stricker, "A new tool for the observation of embryos and other large specimens - confocal theta-fluorescence microscopy," J. Microsc. 179, 1-10 (1995).
[CrossRef]

Swoger, J.

J. Huisken, J. Swoger, F. Del Bene, J. Wittbrodt, and E. H. K. Stelzer, "Optical sectioning deep inside live embryos by selective plane illumination microscopy," Science 305, 1007-1009 (2004).
[CrossRef] [PubMed]

Tannous, T.

Török, P.

Voie, A. H.

A. H. Voie, D. H. Burns, and F. A. Spelman, "Orthogonal-plane fluorescence optical sectioning: three-dimensional imaging of macroscopic biological specimens," J. Microsc. 170, 229-236 (1993).
[CrossRef] [PubMed]

Wang, T. D.

Wilson, T.

Wittbrodt, J.

J. Huisken, J. Swoger, F. Del Bene, J. Wittbrodt, and E. H. K. Stelzer, "Optical sectioning deep inside live embryos by selective plane illumination microscopy," Science 305, 1007-1009 (2004).
[CrossRef] [PubMed]

Wolf, E.

E. H. Linfoot and E. Wolf, "Diffraction images in systems with an annular aperture," Proc. Phys. Soc. B 66, 145-149 (1953).
[CrossRef]

Zhou, H.

M. Gu, C. J. R. Sheppard, and H. Zhou, "Optimization of axial resolution in confocal imaging using annular pupils," Optik 93, 87-90 (1993).

Zsigmondy, R.

H. Siedentopf and R. Zsigmondy, "Uber Sichtbarmachung und Grössenbestimmung ultramikroskopischer Teilchen, mit besonderer Anwendung auf Goldrubingläser," Annalen der Physik 10, 1-39 (1903).

Annalen der Physik

H. Siedentopf and R. Zsigmondy, "Uber Sichtbarmachung und Grössenbestimmung ultramikroskopischer Teilchen, mit besonderer Anwendung auf Goldrubingläser," Annalen der Physik 10, 1-39 (1903).

Appl. Opt.

Experientia

D. M. Maurice, "Cellular membrane activity in the corneal endothelium of the intact eye," Experientia 15, 1094-1095 (1968).
[CrossRef]

Invest. Ophthalmol.

D. M. Maurice, "A scanning slit optical microscope," Invest. Ophthalmol. 13, 1033-1037 (1974).
[PubMed]

J. Microsc.

E. H. K. Stelzer, S. Lindek, S. Albrecht, R. Pick, G. Ritter, N. J. Salmon, and R. Stricker, "A new tool for the observation of embryos and other large specimens - confocal theta-fluorescence microscopy," J. Microsc. 179, 1-10 (1995).
[CrossRef]

A. H. Voie, D. H. Burns, and F. A. Spelman, "Orthogonal-plane fluorescence optical sectioning: three-dimensional imaging of macroscopic biological specimens," J. Microsc. 170, 229-236 (1993).
[CrossRef] [PubMed]

J. Mod. Opt.

C. J. R. Sheppard and M. D. Sharma, "Integrated intensity, and imaging through scattering media," J. Mod. Opt. 48, 1517-1525 (2001).

Ophthalmologica

H. Goldman, "Spaltlampenphotographie und -photometrie," Ophthalmologica 98, 257-270 (1940).
[CrossRef]

Opt. Lett.

Optik

M. Gu, C. J. R. Sheppard, and H. Zhou, "Optimization of axial resolution in confocal imaging using annular pupils," Optik 93, 87-90 (1993).

Proc. Phys. Soc. B

E. H. Linfoot and E. Wolf, "Diffraction images in systems with an annular aperture," Proc. Phys. Soc. B 66, 145-149 (1953).
[CrossRef]

Scanning

X. S. Gan and C. J. R. Sheppard, "Detectability: A new criterion for evaluation of the confocal microscope," Scanning 15, 187-192 (1993).
[CrossRef]

Science

J. Huisken, J. Swoger, F. Del Bene, J. Wittbrodt, and E. H. K. Stelzer, "Optical sectioning deep inside live embryos by selective plane illumination microscopy," Science 305, 1007-1009 (2004).
[CrossRef] [PubMed]

Other

C. J. R. Sheppard, X. Gan, M. Gu, and M. Roy, "Noise in confocal microscopes," in The Handbook of Biological Confocal Microscopy, 2nd edition ed., J. Pawley, ed. (Plenum Press, New York, 1995), pp. 363-370.

M. Born and E. Wolf, Principles of Optics (Pergamon, Oxford, 1959), p. 440.

R. N. Bracewell, The Fourier Transform and its Applications (McGraw-Hill, New York, 1965), p. 253.

C. J. Koester, "Comparison of optical sectioning methods: The scanning slit confocal microscope," in Handbook of Confocal Microscopy, J. Pawley, ed. (Plenum Press, New York, 1990).

Supplementary Material (2)

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Figures (10)

Fig. 1.
Fig. 1.

Schematic diagram of a confocal system with D-shaped pupils.

Fig. 2.
Fig. 2.

The intensity point spread function for a D-shaped pupil for d=0.1 (Media 1). As defocus is increased a Poisson spot develops on the optical axis.

Fig. 3.
Fig. 3.

Cross-sections through the intensity point spread function for a D-shaped pupil for d=0.1: (a) x-z section, (b) y-z section.

Fig. 4.
Fig. 4.

The pupil function P(t) for different values of d.

Fig. 5.
Fig. 5.

The intensity along the axis for a D-shaped pupil shown as a log-log plot.

Fig. 6.
Fig. 6.

The intensity point spread function for a confocal microscope with two D-shaped pupils and a point detector, d=0.1 (Media 2). The Poisson spots from the two point spread functions overlap.

Fig. 7.
Fig. 7.

Cross-section x-z through the intensity point spread function for a confocal microscope with two D-shaped pupils and a point detector, d=0.1.

Fig. 8.
Fig. 8.

Half-width at half-maximum (HWHM) of the intensity point spread function for a confocal microscope with two D-shaped pupils and a point detector in the vx , vy and u directions.

Fig. 9.
Fig. 9.

The integrated intensity I int(u) for a confocal microscope with two D-shaped pupils and a point detector shown as a log-log plot.

Fig. 10.
Fig. 10.

Signal/Background for a confocal microscope with two D-shaped pupils and a point detector as a function of d.

Equations (4)

Equations on this page are rendered with MathJax. Learn more.

U ( v x , v y , u ) = 1 π pupil P ( ρ x , ρ y ) exp { i [ v x ρ x + v y ρ y 1 2 u ( ρ x 2 + ρ y 2 ) ] } d ρ x d ρ y .
P ( t ) = 0 2 π P ( ρ , ϕ ) d ϕ = 2 arccos ( d t ) .
U ( 0 ) = 1 π [ arccos d d ( 1 d 2 ) 1 / 2 ]
U = 1 2 π d 2 1 P ( t ) e iut / 2 d t .

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