Abstract

Optical projection tomography (OPT) requires the depth of field (DOF) of the lens to cover at least half of the sample. There is a trade-off between obtaining high resolution with a high-NA lens and obtaining large DOF with a low-NA lens. The DOF of a high-NA objective lens can be extended by scanning its focal plane through the sample. We call this extended DOF image a “pseudoprojection.” Images reconstructed from these pseudoprojections have isometric resolution, which can be the same as the lateral resolution of the high-NA objective. The focal scanning method produces an over 10× improvement in OPT resolution.

© 2010 Optical Society of America

Full Article  |  PDF Article

References

  • View by:
  • |
  • |
  • |

  1. J. Sharpe, U. Ahlgren, P. Perry, B. Hill, A. Ross, J. Hecksher-Sorensen, R. Baldock, and D. Davidson, Science 296, 541 (2002).
    [CrossRef] [PubMed]
  2. J. R. Walls, L. Coultas, J. Rossant, and R. M. Henkelman, PLoS ONE 3, e2853 (2008).
    [CrossRef] [PubMed]
  3. K. Lee, J. Avondo, H. Morrison, L. Blot, M. Stark, J. Sharpe, A. Bangham, and E. Coen, Plant Cell Physiol. 18, 2145(2006).
  4. T. Alanentalo, A. Asayesh, H. Morrison, C. E. Loren, D. Holmberg, J. Sharpe, and U. Ahlgren, Nat. Meth. 3, 31(2007).
    [CrossRef]
  5. J. R. Walls, J. G. Sled, J. Sharpe, and R. M. Henkelman, Phys. Med. Biol. 52, 2775 (2007).
    [CrossRef] [PubMed]
  6. G. Hausler, Opt. Commun. 6, 38 (1972).
    [CrossRef]
  7. M. Fauver, E. J. Seibel, J. R. Rahn, M. G. Meyer, F. W. Patten, T. Neumann, and A. C. Nelson, Opt. Express 13, 4210 (2005).
    [CrossRef] [PubMed]
  8. Q. Miao, J. R. Rahn, A. Tourovskaia, M. G. Meyer, T. Neumann, A. C. Nelson, and E. J. Seibel, J. Biomed. Opt. 14, 064035 (2009).
    [CrossRef]
  9. J. W. Goodman, Introduction to Fourier Optics, 3rd ed. (Roberts, 2005).
  10. A. C. Kak and M. Slaney, Principles of Computerized Tomographic Imaging (IEEE, 1988).
  11. H. S. Sakhalkar, M. Dewhirst, T. Oliver, Y. Cao, and M. Oldham, Phys. Med. Biol. 52, 2035 (2007).
    [CrossRef] [PubMed]

2009 (1)

Q. Miao, J. R. Rahn, A. Tourovskaia, M. G. Meyer, T. Neumann, A. C. Nelson, and E. J. Seibel, J. Biomed. Opt. 14, 064035 (2009).
[CrossRef]

2008 (1)

J. R. Walls, L. Coultas, J. Rossant, and R. M. Henkelman, PLoS ONE 3, e2853 (2008).
[CrossRef] [PubMed]

2007 (3)

T. Alanentalo, A. Asayesh, H. Morrison, C. E. Loren, D. Holmberg, J. Sharpe, and U. Ahlgren, Nat. Meth. 3, 31(2007).
[CrossRef]

J. R. Walls, J. G. Sled, J. Sharpe, and R. M. Henkelman, Phys. Med. Biol. 52, 2775 (2007).
[CrossRef] [PubMed]

H. S. Sakhalkar, M. Dewhirst, T. Oliver, Y. Cao, and M. Oldham, Phys. Med. Biol. 52, 2035 (2007).
[CrossRef] [PubMed]

2006 (1)

K. Lee, J. Avondo, H. Morrison, L. Blot, M. Stark, J. Sharpe, A. Bangham, and E. Coen, Plant Cell Physiol. 18, 2145(2006).

2005 (2)

2002 (1)

J. Sharpe, U. Ahlgren, P. Perry, B. Hill, A. Ross, J. Hecksher-Sorensen, R. Baldock, and D. Davidson, Science 296, 541 (2002).
[CrossRef] [PubMed]

1988 (1)

A. C. Kak and M. Slaney, Principles of Computerized Tomographic Imaging (IEEE, 1988).

1972 (1)

G. Hausler, Opt. Commun. 6, 38 (1972).
[CrossRef]

Ahlgren, U.

T. Alanentalo, A. Asayesh, H. Morrison, C. E. Loren, D. Holmberg, J. Sharpe, and U. Ahlgren, Nat. Meth. 3, 31(2007).
[CrossRef]

J. Sharpe, U. Ahlgren, P. Perry, B. Hill, A. Ross, J. Hecksher-Sorensen, R. Baldock, and D. Davidson, Science 296, 541 (2002).
[CrossRef] [PubMed]

Alanentalo, T.

T. Alanentalo, A. Asayesh, H. Morrison, C. E. Loren, D. Holmberg, J. Sharpe, and U. Ahlgren, Nat. Meth. 3, 31(2007).
[CrossRef]

Asayesh, A.

T. Alanentalo, A. Asayesh, H. Morrison, C. E. Loren, D. Holmberg, J. Sharpe, and U. Ahlgren, Nat. Meth. 3, 31(2007).
[CrossRef]

Avondo, J.

K. Lee, J. Avondo, H. Morrison, L. Blot, M. Stark, J. Sharpe, A. Bangham, and E. Coen, Plant Cell Physiol. 18, 2145(2006).

Baldock, R.

J. Sharpe, U. Ahlgren, P. Perry, B. Hill, A. Ross, J. Hecksher-Sorensen, R. Baldock, and D. Davidson, Science 296, 541 (2002).
[CrossRef] [PubMed]

Bangham, A.

K. Lee, J. Avondo, H. Morrison, L. Blot, M. Stark, J. Sharpe, A. Bangham, and E. Coen, Plant Cell Physiol. 18, 2145(2006).

Blot, L.

K. Lee, J. Avondo, H. Morrison, L. Blot, M. Stark, J. Sharpe, A. Bangham, and E. Coen, Plant Cell Physiol. 18, 2145(2006).

Cao, Y.

H. S. Sakhalkar, M. Dewhirst, T. Oliver, Y. Cao, and M. Oldham, Phys. Med. Biol. 52, 2035 (2007).
[CrossRef] [PubMed]

Coen, E.

K. Lee, J. Avondo, H. Morrison, L. Blot, M. Stark, J. Sharpe, A. Bangham, and E. Coen, Plant Cell Physiol. 18, 2145(2006).

Coultas, L.

J. R. Walls, L. Coultas, J. Rossant, and R. M. Henkelman, PLoS ONE 3, e2853 (2008).
[CrossRef] [PubMed]

Davidson, D.

J. Sharpe, U. Ahlgren, P. Perry, B. Hill, A. Ross, J. Hecksher-Sorensen, R. Baldock, and D. Davidson, Science 296, 541 (2002).
[CrossRef] [PubMed]

Dewhirst, M.

H. S. Sakhalkar, M. Dewhirst, T. Oliver, Y. Cao, and M. Oldham, Phys. Med. Biol. 52, 2035 (2007).
[CrossRef] [PubMed]

Fauver, M.

Goodman, J. W.

J. W. Goodman, Introduction to Fourier Optics, 3rd ed. (Roberts, 2005).

Hausler, G.

G. Hausler, Opt. Commun. 6, 38 (1972).
[CrossRef]

Hecksher-Sorensen, J.

J. Sharpe, U. Ahlgren, P. Perry, B. Hill, A. Ross, J. Hecksher-Sorensen, R. Baldock, and D. Davidson, Science 296, 541 (2002).
[CrossRef] [PubMed]

Henkelman, R. M.

J. R. Walls, L. Coultas, J. Rossant, and R. M. Henkelman, PLoS ONE 3, e2853 (2008).
[CrossRef] [PubMed]

J. R. Walls, J. G. Sled, J. Sharpe, and R. M. Henkelman, Phys. Med. Biol. 52, 2775 (2007).
[CrossRef] [PubMed]

Hill, B.

J. Sharpe, U. Ahlgren, P. Perry, B. Hill, A. Ross, J. Hecksher-Sorensen, R. Baldock, and D. Davidson, Science 296, 541 (2002).
[CrossRef] [PubMed]

Holmberg, D.

T. Alanentalo, A. Asayesh, H. Morrison, C. E. Loren, D. Holmberg, J. Sharpe, and U. Ahlgren, Nat. Meth. 3, 31(2007).
[CrossRef]

Kak, A. C.

A. C. Kak and M. Slaney, Principles of Computerized Tomographic Imaging (IEEE, 1988).

Lee, K.

K. Lee, J. Avondo, H. Morrison, L. Blot, M. Stark, J. Sharpe, A. Bangham, and E. Coen, Plant Cell Physiol. 18, 2145(2006).

Loren, C. E.

T. Alanentalo, A. Asayesh, H. Morrison, C. E. Loren, D. Holmberg, J. Sharpe, and U. Ahlgren, Nat. Meth. 3, 31(2007).
[CrossRef]

Meyer, M. G.

Q. Miao, J. R. Rahn, A. Tourovskaia, M. G. Meyer, T. Neumann, A. C. Nelson, and E. J. Seibel, J. Biomed. Opt. 14, 064035 (2009).
[CrossRef]

M. Fauver, E. J. Seibel, J. R. Rahn, M. G. Meyer, F. W. Patten, T. Neumann, and A. C. Nelson, Opt. Express 13, 4210 (2005).
[CrossRef] [PubMed]

Miao, Q.

Q. Miao, J. R. Rahn, A. Tourovskaia, M. G. Meyer, T. Neumann, A. C. Nelson, and E. J. Seibel, J. Biomed. Opt. 14, 064035 (2009).
[CrossRef]

Morrison, H.

T. Alanentalo, A. Asayesh, H. Morrison, C. E. Loren, D. Holmberg, J. Sharpe, and U. Ahlgren, Nat. Meth. 3, 31(2007).
[CrossRef]

K. Lee, J. Avondo, H. Morrison, L. Blot, M. Stark, J. Sharpe, A. Bangham, and E. Coen, Plant Cell Physiol. 18, 2145(2006).

Nelson, A. C.

Q. Miao, J. R. Rahn, A. Tourovskaia, M. G. Meyer, T. Neumann, A. C. Nelson, and E. J. Seibel, J. Biomed. Opt. 14, 064035 (2009).
[CrossRef]

M. Fauver, E. J. Seibel, J. R. Rahn, M. G. Meyer, F. W. Patten, T. Neumann, and A. C. Nelson, Opt. Express 13, 4210 (2005).
[CrossRef] [PubMed]

Neumann, T.

Q. Miao, J. R. Rahn, A. Tourovskaia, M. G. Meyer, T. Neumann, A. C. Nelson, and E. J. Seibel, J. Biomed. Opt. 14, 064035 (2009).
[CrossRef]

M. Fauver, E. J. Seibel, J. R. Rahn, M. G. Meyer, F. W. Patten, T. Neumann, and A. C. Nelson, Opt. Express 13, 4210 (2005).
[CrossRef] [PubMed]

Oldham, M.

H. S. Sakhalkar, M. Dewhirst, T. Oliver, Y. Cao, and M. Oldham, Phys. Med. Biol. 52, 2035 (2007).
[CrossRef] [PubMed]

Oliver, T.

H. S. Sakhalkar, M. Dewhirst, T. Oliver, Y. Cao, and M. Oldham, Phys. Med. Biol. 52, 2035 (2007).
[CrossRef] [PubMed]

Patten, F. W.

Perry, P.

J. Sharpe, U. Ahlgren, P. Perry, B. Hill, A. Ross, J. Hecksher-Sorensen, R. Baldock, and D. Davidson, Science 296, 541 (2002).
[CrossRef] [PubMed]

Rahn, J. R.

Q. Miao, J. R. Rahn, A. Tourovskaia, M. G. Meyer, T. Neumann, A. C. Nelson, and E. J. Seibel, J. Biomed. Opt. 14, 064035 (2009).
[CrossRef]

M. Fauver, E. J. Seibel, J. R. Rahn, M. G. Meyer, F. W. Patten, T. Neumann, and A. C. Nelson, Opt. Express 13, 4210 (2005).
[CrossRef] [PubMed]

Ross, A.

J. Sharpe, U. Ahlgren, P. Perry, B. Hill, A. Ross, J. Hecksher-Sorensen, R. Baldock, and D. Davidson, Science 296, 541 (2002).
[CrossRef] [PubMed]

Rossant, J.

J. R. Walls, L. Coultas, J. Rossant, and R. M. Henkelman, PLoS ONE 3, e2853 (2008).
[CrossRef] [PubMed]

Sakhalkar, H. S.

H. S. Sakhalkar, M. Dewhirst, T. Oliver, Y. Cao, and M. Oldham, Phys. Med. Biol. 52, 2035 (2007).
[CrossRef] [PubMed]

Seibel, E. J.

Q. Miao, J. R. Rahn, A. Tourovskaia, M. G. Meyer, T. Neumann, A. C. Nelson, and E. J. Seibel, J. Biomed. Opt. 14, 064035 (2009).
[CrossRef]

M. Fauver, E. J. Seibel, J. R. Rahn, M. G. Meyer, F. W. Patten, T. Neumann, and A. C. Nelson, Opt. Express 13, 4210 (2005).
[CrossRef] [PubMed]

Sharpe, J.

J. R. Walls, J. G. Sled, J. Sharpe, and R. M. Henkelman, Phys. Med. Biol. 52, 2775 (2007).
[CrossRef] [PubMed]

T. Alanentalo, A. Asayesh, H. Morrison, C. E. Loren, D. Holmberg, J. Sharpe, and U. Ahlgren, Nat. Meth. 3, 31(2007).
[CrossRef]

K. Lee, J. Avondo, H. Morrison, L. Blot, M. Stark, J. Sharpe, A. Bangham, and E. Coen, Plant Cell Physiol. 18, 2145(2006).

J. Sharpe, U. Ahlgren, P. Perry, B. Hill, A. Ross, J. Hecksher-Sorensen, R. Baldock, and D. Davidson, Science 296, 541 (2002).
[CrossRef] [PubMed]

Slaney, M.

A. C. Kak and M. Slaney, Principles of Computerized Tomographic Imaging (IEEE, 1988).

Sled, J. G.

J. R. Walls, J. G. Sled, J. Sharpe, and R. M. Henkelman, Phys. Med. Biol. 52, 2775 (2007).
[CrossRef] [PubMed]

Stark, M.

K. Lee, J. Avondo, H. Morrison, L. Blot, M. Stark, J. Sharpe, A. Bangham, and E. Coen, Plant Cell Physiol. 18, 2145(2006).

Tourovskaia, A.

Q. Miao, J. R. Rahn, A. Tourovskaia, M. G. Meyer, T. Neumann, A. C. Nelson, and E. J. Seibel, J. Biomed. Opt. 14, 064035 (2009).
[CrossRef]

Walls, J. R.

J. R. Walls, L. Coultas, J. Rossant, and R. M. Henkelman, PLoS ONE 3, e2853 (2008).
[CrossRef] [PubMed]

J. R. Walls, J. G. Sled, J. Sharpe, and R. M. Henkelman, Phys. Med. Biol. 52, 2775 (2007).
[CrossRef] [PubMed]

J. Biomed. Opt. (1)

Q. Miao, J. R. Rahn, A. Tourovskaia, M. G. Meyer, T. Neumann, A. C. Nelson, and E. J. Seibel, J. Biomed. Opt. 14, 064035 (2009).
[CrossRef]

Nat. Meth. (1)

T. Alanentalo, A. Asayesh, H. Morrison, C. E. Loren, D. Holmberg, J. Sharpe, and U. Ahlgren, Nat. Meth. 3, 31(2007).
[CrossRef]

Opt. Commun. (1)

G. Hausler, Opt. Commun. 6, 38 (1972).
[CrossRef]

Opt. Express (1)

Phys. Med. Biol. (2)

J. R. Walls, J. G. Sled, J. Sharpe, and R. M. Henkelman, Phys. Med. Biol. 52, 2775 (2007).
[CrossRef] [PubMed]

H. S. Sakhalkar, M. Dewhirst, T. Oliver, Y. Cao, and M. Oldham, Phys. Med. Biol. 52, 2035 (2007).
[CrossRef] [PubMed]

Plant Cell Physiol. (1)

K. Lee, J. Avondo, H. Morrison, L. Blot, M. Stark, J. Sharpe, A. Bangham, and E. Coen, Plant Cell Physiol. 18, 2145(2006).

PLoS ONE (1)

J. R. Walls, L. Coultas, J. Rossant, and R. M. Henkelman, PLoS ONE 3, e2853 (2008).
[CrossRef] [PubMed]

Science (1)

J. Sharpe, U. Ahlgren, P. Perry, B. Hill, A. Ross, J. Hecksher-Sorensen, R. Baldock, and D. Davidson, Science 296, 541 (2002).
[CrossRef] [PubMed]

Other (2)

J. W. Goodman, Introduction to Fourier Optics, 3rd ed. (Roberts, 2005).

A. C. Kak and M. Slaney, Principles of Computerized Tomographic Imaging (IEEE, 1988).

Cited By

OSA participates in CrossRef's Cited-By Linking service. Citing articles from OSA journals and other participating publishers are listed here.

Alert me when this article is cited.


Figures (5)

Fig. 1
Fig. 1

Schematic of OPT microscope. The Z axis is along the tube axis; Y is along the optical axis of the objective.

Fig. 2
Fig. 2

Schematic of optical system when sample is rotated θ ° . When θ = 0 , the X and Y coordinates are the same as those shown in Fig. 1.

Fig. 3
Fig. 3

(a) h , (b) h P , and (c) h for an 100 × objective with NA 1.3. h is the original PSF of the objective, h P is the projection of h along the optical axis, and h is the PSF of the system and is calculated from the projection image h P . All coordinates shown are the same as those shown in Fig. 2. (c) The axial resolution is the same as the lateral resolution.

Fig. 4
Fig. 4

Comparison of the PSF in the axial section between (a) conventional OPT and (b) the scanning method. The coordinates are the same as those in Fig. 1; the Y axis is along the optical axis.

Fig. 5
Fig. 5

3D reconstruction of clusters of muntjac cells stained with hematoxylin.

Equations (5)

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

P θ ( t ) = s I ( s , t ) d s = ( t , s ) O ( s , t ) ( s h ( s s , t t ) d s ) d t d s = ( t , s ) O ( s , t ) h p ( t t ) d t d s = t ( s O ( s , t ) d s ) h p ( t t ) d t = t ( O p ( t ) h p ( t t ) d t = O p ( t ) * h p ( t ) ,
O R ( x , y , z ) = 0 π S θ ( w ) | w | exp ( j 2 π w t 1 ) d w d θ ,
S θ ( w ) = P θ ( t 1 , z ) exp ( j 2 π w t 1 ) d t 1 = O p ( t 1 , z ) * h p ( t 1 , z ) exp ( j 2 π w t 1 ) d t 1 ,
O R ( x , y , z ) = O ( x , y , z ) * h ( x , y , z ) ,
h ( x , y , z ) = 0 π ( h p ( t 1 , z ) exp ( j 2 π w t 1 ) d t 1 ) | w | exp ( j 2 π w t 1 ) d w d θ .

Metrics