Abstract

The standard microscope architecture around which confocal microscopes are built imposes fundamental restrictions on the speed with which images (three-dimensional data sets) can be obtained. Commercially available slit scanning confocal microscopes are able to produce optically sectioned images at frame rates well in excess of 100 Hz. However only the focal (xy) plane can be imaged at this speed. To image a volume specimen it is necessary to physically change the distance between the objective lens and the specimen. This refocusing process is often necessarily slow and represents a bottleneck to the speed of image acquisition. We describe the construction of a slit scanning confocal microscope based on what we know to be a novel microscope architecture, which permits images of other planes and, particular, the meridional (xz) plane to be acquired in real time.

© 2009 Optical Society of America

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References

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    [CrossRef]
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2009

M. Murayama, E. Pérez-Garci, T. Nevian, T. Bock, W. Senn, and M. E. Larkum, Nature 457, 1137 (2009).
[CrossRef] [PubMed]

2008

E. Botcherby, R. Juškaitis, M. Booth, and T. Wilson, Opt. Commun. 281, 880 (2008).
[CrossRef]

2007

2006

D. Hughes, U. Tirlapur, R. Field, and Z. Cui, J. Membr. Sci. 280, 124 (2006).
[CrossRef]

R. Wolleschensky, B. Zimmermann, and M. Kempe, J. Biomed. Opt. 11, 064011 (2006).
[CrossRef]

1992

G. J. Brakenhoff and K. Visscher, J. Microsc. 165, 139 (1992).
[CrossRef]

1990

D. H. Burns, R. B. Hatangadi, and F. A. Spelman, Scanning 12, 156 (1990).
[CrossRef]

1988

C. J. R. Sheppard and X. Q. Moa, J. Mod. Opt. 35, 1169 (1988).
[CrossRef]

Bock, T.

M. Murayama, E. Pérez-Garci, T. Nevian, T. Bock, W. Senn, and M. E. Larkum, Nature 457, 1137 (2009).
[CrossRef] [PubMed]

Booth, M.

E. Botcherby, R. Juškaitis, M. Booth, and T. Wilson, Opt. Commun. 281, 880 (2008).
[CrossRef]

E. Botcherby, R. Juškaitis, M. Booth, and T. Wilson, Opt. Lett. 32, 2007 (2007).
[CrossRef] [PubMed]

Botcherby, E.

E. Botcherby, R. Juškaitis, M. Booth, and T. Wilson, Opt. Commun. 281, 880 (2008).
[CrossRef]

E. Botcherby, R. Juškaitis, M. Booth, and T. Wilson, Opt. Lett. 32, 2007 (2007).
[CrossRef] [PubMed]

Brakenhoff, G. J.

G. J. Brakenhoff and K. Visscher, J. Microsc. 165, 139 (1992).
[CrossRef]

Burns, D. H.

D. H. Burns, R. B. Hatangadi, and F. A. Spelman, Scanning 12, 156 (1990).
[CrossRef]

Cui, Z.

D. Hughes, U. Tirlapur, R. Field, and Z. Cui, J. Membr. Sci. 280, 124 (2006).
[CrossRef]

Field, R.

D. Hughes, U. Tirlapur, R. Field, and Z. Cui, J. Membr. Sci. 280, 124 (2006).
[CrossRef]

Hatangadi, R. B.

D. H. Burns, R. B. Hatangadi, and F. A. Spelman, Scanning 12, 156 (1990).
[CrossRef]

Hughes, D.

D. Hughes, U. Tirlapur, R. Field, and Z. Cui, J. Membr. Sci. 280, 124 (2006).
[CrossRef]

Juškaitis, R.

E. Botcherby, R. Juškaitis, M. Booth, and T. Wilson, Opt. Commun. 281, 880 (2008).
[CrossRef]

E. Botcherby, R. Juškaitis, M. Booth, and T. Wilson, Opt. Lett. 32, 2007 (2007).
[CrossRef] [PubMed]

Kempe, M.

R. Wolleschensky, B. Zimmermann, and M. Kempe, J. Biomed. Opt. 11, 064011 (2006).
[CrossRef]

Larkum, M. E.

M. Murayama, E. Pérez-Garci, T. Nevian, T. Bock, W. Senn, and M. E. Larkum, Nature 457, 1137 (2009).
[CrossRef] [PubMed]

Moa, X. Q.

C. J. R. Sheppard and X. Q. Moa, J. Mod. Opt. 35, 1169 (1988).
[CrossRef]

Murayama, M.

M. Murayama, E. Pérez-Garci, T. Nevian, T. Bock, W. Senn, and M. E. Larkum, Nature 457, 1137 (2009).
[CrossRef] [PubMed]

Nevian, T.

M. Murayama, E. Pérez-Garci, T. Nevian, T. Bock, W. Senn, and M. E. Larkum, Nature 457, 1137 (2009).
[CrossRef] [PubMed]

Pérez-Garci, E.

M. Murayama, E. Pérez-Garci, T. Nevian, T. Bock, W. Senn, and M. E. Larkum, Nature 457, 1137 (2009).
[CrossRef] [PubMed]

Senn, W.

M. Murayama, E. Pérez-Garci, T. Nevian, T. Bock, W. Senn, and M. E. Larkum, Nature 457, 1137 (2009).
[CrossRef] [PubMed]

Sheppard, C. J. R.

C. J. R. Sheppard and X. Q. Moa, J. Mod. Opt. 35, 1169 (1988).
[CrossRef]

Spelman, F. A.

D. H. Burns, R. B. Hatangadi, and F. A. Spelman, Scanning 12, 156 (1990).
[CrossRef]

Tirlapur, U.

D. Hughes, U. Tirlapur, R. Field, and Z. Cui, J. Membr. Sci. 280, 124 (2006).
[CrossRef]

Visscher, K.

G. J. Brakenhoff and K. Visscher, J. Microsc. 165, 139 (1992).
[CrossRef]

Wilson, T.

E. Botcherby, R. Juškaitis, M. Booth, and T. Wilson, Opt. Commun. 281, 880 (2008).
[CrossRef]

E. Botcherby, R. Juškaitis, M. Booth, and T. Wilson, Opt. Lett. 32, 2007 (2007).
[CrossRef] [PubMed]

Wolleschensky, R.

R. Wolleschensky, B. Zimmermann, and M. Kempe, J. Biomed. Opt. 11, 064011 (2006).
[CrossRef]

Zimmermann, B.

R. Wolleschensky, B. Zimmermann, and M. Kempe, J. Biomed. Opt. 11, 064011 (2006).
[CrossRef]

J. Biomed. Opt.

R. Wolleschensky, B. Zimmermann, and M. Kempe, J. Biomed. Opt. 11, 064011 (2006).
[CrossRef]

J. Membr. Sci.

D. Hughes, U. Tirlapur, R. Field, and Z. Cui, J. Membr. Sci. 280, 124 (2006).
[CrossRef]

J. Microsc.

G. J. Brakenhoff and K. Visscher, J. Microsc. 165, 139 (1992).
[CrossRef]

J. Mod. Opt.

C. J. R. Sheppard and X. Q. Moa, J. Mod. Opt. 35, 1169 (1988).
[CrossRef]

Nature

M. Murayama, E. Pérez-Garci, T. Nevian, T. Bock, W. Senn, and M. E. Larkum, Nature 457, 1137 (2009).
[CrossRef] [PubMed]

Opt. Commun.

E. Botcherby, R. Juškaitis, M. Booth, and T. Wilson, Opt. Commun. 281, 880 (2008).
[CrossRef]

Opt. Lett.

Scanning

D. H. Burns, R. B. Hatangadi, and F. A. Spelman, Scanning 12, 156 (1990).
[CrossRef]

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Figures (2)

Fig. 1
Fig. 1

Slit scanning confocal microscope that images the MP directly with a CCD camera. Inset, line information from different specimen depths is imaged onto different rows of pixels during each frame.

Fig. 2
Fig. 2

Image of the parts of a fluorescent pollen grain lying in the MP of our slit scanning confocal microscope. These are frames from a real-time movie obtained of the specimen.

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