Abstract

We describe an angular multiplexing technique for optical projection tomography that improves resolution, signal-to-noise ratio, and imaging speed by ameliorating the trade-off between spatial resolution and depth of field and improving the light collection efficiency. Here we demonstrate that imaging at two orthogonal angular projections simultaneously, focused on shifted planes in the sample, improves the average spatial resolution by 20% and the light collection efficiency by a factor of 4, thereby enabling increased acquisition speed and reduced light dose.

© 2013 Optical Society of America

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  1. J. Sharpe, U. Ahlgren, P. Perry, B. Hill, A. Ross, J. Hecksher-Sorensen, R. Baldock, and D. Davidson, Science 296, 541 (2002).
    [CrossRef]
  2. R. Lorbeer, M. Heidrich, C. Lorbeer, D. F. R. Ojeda, G. Bicker, H. Meyer, and A. Heisterkamp, Opt. Express 19, 5419 (2011).
    [CrossRef]
  3. J. Huisken, J. Swoger, F. Del Bene, J. Wittbrodt, and E. H. K. Stelzer, Science 305, 1007 (2004).
    [CrossRef]
  4. H. U. Dodt, U. Leischner, A. Schierloh, N. Jahrling, C. P. Mauch, K. Deininger, J. M. Deussing, M. Eder, W. Zieglgansberger, and K. Becker, Nat. Methods 4, 331 (2007).
    [CrossRef]
  5. N. Krstajic and S. J. Doran, Phys. Med. Biol. 51, 2055 (2006).
    [CrossRef]
  6. A. C. Kak and M. Slaney, Principles of Computerized Tomographic Imaging (SIAM, 1988).
  7. M. A. Haidekker, Comput. Methods Programs Biomed. 80, 225 (2005).
    [CrossRef]
  8. J. McGinty, K. B. Tahir, R. Laine, C. B. Talbot, C. Dunsby, M. A. A. Neil, L. Quintana, J. Swoger, J. Sharpe, and P. M. W. French, J. Biophoton. 1, 390 (2008).
    [CrossRef]
  9. T. Alanentalo, A. Asayesh, H. Morrison, C. E. Lorén, D. Holmberg, J. Sharpe, and U. Ahlgren, Nat. Methods 4, 31 (2007).
    [CrossRef]
  10. U. J. Birk, M. Rieckher, N. Konstantinides, A. Darrell, A. Sarasa-Renedo, H. Meyer, N. Tavernarakis, and J. Ripoll, Biomed. Opt. Express 1, 87 (2010).
    [CrossRef]
  11. A. Bassi, L. Fieramonti, C. D’Andrea, M. Mione, and G. Valentini, J. Biomed. Opt. 16, 100502 (2011).
    [CrossRef]
  12. J. R. Walls, J. G. Sled, J. Sharpe, and R. M. Henkelman, Phys. Med. Biol. 50, 4645 (2005).
    [CrossRef]
  13. J. R. Walls, J. G. Sled, J. Sharpe, and R. M. Henkelman, Phys. Med. Biol. 52, 2775 (2007).
    [CrossRef]
  14. L. Chen, J. McGinty, H. B. Taylor, L. Bugeon, J. R. Lamb, M. J. Dallman, and P. M. W. French, Opt. Express 20, 7323 (2012).
    [CrossRef]
  15. Q. Miao, J. Hayenga, M. G. Meyer, T. Neumann, A. C. Nelson, and E. J. Seibel, Opt. Lett. 35, 3363 (2010).
    [CrossRef]

2012 (1)

2011 (2)

A. Bassi, L. Fieramonti, C. D’Andrea, M. Mione, and G. Valentini, J. Biomed. Opt. 16, 100502 (2011).
[CrossRef]

R. Lorbeer, M. Heidrich, C. Lorbeer, D. F. R. Ojeda, G. Bicker, H. Meyer, and A. Heisterkamp, Opt. Express 19, 5419 (2011).
[CrossRef]

2010 (2)

2008 (1)

J. McGinty, K. B. Tahir, R. Laine, C. B. Talbot, C. Dunsby, M. A. A. Neil, L. Quintana, J. Swoger, J. Sharpe, and P. M. W. French, J. Biophoton. 1, 390 (2008).
[CrossRef]

2007 (3)

T. Alanentalo, A. Asayesh, H. Morrison, C. E. Lorén, D. Holmberg, J. Sharpe, and U. Ahlgren, Nat. Methods 4, 31 (2007).
[CrossRef]

H. U. Dodt, U. Leischner, A. Schierloh, N. Jahrling, C. P. Mauch, K. Deininger, J. M. Deussing, M. Eder, W. Zieglgansberger, and K. Becker, Nat. Methods 4, 331 (2007).
[CrossRef]

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

2006 (1)

N. Krstajic and S. J. Doran, Phys. Med. Biol. 51, 2055 (2006).
[CrossRef]

2005 (2)

M. A. Haidekker, Comput. Methods Programs Biomed. 80, 225 (2005).
[CrossRef]

J. R. Walls, J. G. Sled, J. Sharpe, and R. M. Henkelman, Phys. Med. Biol. 50, 4645 (2005).
[CrossRef]

2004 (1)

J. Huisken, J. Swoger, F. Del Bene, J. Wittbrodt, and E. H. K. Stelzer, Science 305, 1007 (2004).
[CrossRef]

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]

Ahlgren, U.

T. Alanentalo, A. Asayesh, H. Morrison, C. E. Lorén, D. Holmberg, J. Sharpe, and U. Ahlgren, Nat. Methods 4, 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]

Alanentalo, T.

T. Alanentalo, A. Asayesh, H. Morrison, C. E. Lorén, D. Holmberg, J. Sharpe, and U. Ahlgren, Nat. Methods 4, 31 (2007).
[CrossRef]

Asayesh, A.

T. Alanentalo, A. Asayesh, H. Morrison, C. E. Lorén, D. Holmberg, J. Sharpe, and U. Ahlgren, Nat. Methods 4, 31 (2007).
[CrossRef]

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]

Bassi, A.

A. Bassi, L. Fieramonti, C. D’Andrea, M. Mione, and G. Valentini, J. Biomed. Opt. 16, 100502 (2011).
[CrossRef]

Becker, K.

H. U. Dodt, U. Leischner, A. Schierloh, N. Jahrling, C. P. Mauch, K. Deininger, J. M. Deussing, M. Eder, W. Zieglgansberger, and K. Becker, Nat. Methods 4, 331 (2007).
[CrossRef]

Bicker, G.

Birk, U. J.

Bugeon, L.

Chen, L.

D’Andrea, C.

A. Bassi, L. Fieramonti, C. D’Andrea, M. Mione, and G. Valentini, J. Biomed. Opt. 16, 100502 (2011).
[CrossRef]

Dallman, M. J.

Darrell, A.

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]

Deininger, K.

H. U. Dodt, U. Leischner, A. Schierloh, N. Jahrling, C. P. Mauch, K. Deininger, J. M. Deussing, M. Eder, W. Zieglgansberger, and K. Becker, Nat. Methods 4, 331 (2007).
[CrossRef]

Del Bene, F.

J. Huisken, J. Swoger, F. Del Bene, J. Wittbrodt, and E. H. K. Stelzer, Science 305, 1007 (2004).
[CrossRef]

Deussing, J. M.

H. U. Dodt, U. Leischner, A. Schierloh, N. Jahrling, C. P. Mauch, K. Deininger, J. M. Deussing, M. Eder, W. Zieglgansberger, and K. Becker, Nat. Methods 4, 331 (2007).
[CrossRef]

Dodt, H. U.

H. U. Dodt, U. Leischner, A. Schierloh, N. Jahrling, C. P. Mauch, K. Deininger, J. M. Deussing, M. Eder, W. Zieglgansberger, and K. Becker, Nat. Methods 4, 331 (2007).
[CrossRef]

Doran, S. J.

N. Krstajic and S. J. Doran, Phys. Med. Biol. 51, 2055 (2006).
[CrossRef]

Dunsby, C.

J. McGinty, K. B. Tahir, R. Laine, C. B. Talbot, C. Dunsby, M. A. A. Neil, L. Quintana, J. Swoger, J. Sharpe, and P. M. W. French, J. Biophoton. 1, 390 (2008).
[CrossRef]

Eder, M.

H. U. Dodt, U. Leischner, A. Schierloh, N. Jahrling, C. P. Mauch, K. Deininger, J. M. Deussing, M. Eder, W. Zieglgansberger, and K. Becker, Nat. Methods 4, 331 (2007).
[CrossRef]

Fieramonti, L.

A. Bassi, L. Fieramonti, C. D’Andrea, M. Mione, and G. Valentini, J. Biomed. Opt. 16, 100502 (2011).
[CrossRef]

French, P. M. W.

L. Chen, J. McGinty, H. B. Taylor, L. Bugeon, J. R. Lamb, M. J. Dallman, and P. M. W. French, Opt. Express 20, 7323 (2012).
[CrossRef]

J. McGinty, K. B. Tahir, R. Laine, C. B. Talbot, C. Dunsby, M. A. A. Neil, L. Quintana, J. Swoger, J. Sharpe, and P. M. W. French, J. Biophoton. 1, 390 (2008).
[CrossRef]

Haidekker, M. A.

M. A. Haidekker, Comput. Methods Programs Biomed. 80, 225 (2005).
[CrossRef]

Hayenga, J.

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]

Heidrich, M.

Heisterkamp, A.

Henkelman, R. M.

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

J. R. Walls, J. G. Sled, J. Sharpe, and R. M. Henkelman, Phys. Med. Biol. 50, 4645 (2005).
[CrossRef]

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]

Holmberg, D.

T. Alanentalo, A. Asayesh, H. Morrison, C. E. Lorén, D. Holmberg, J. Sharpe, and U. Ahlgren, Nat. Methods 4, 31 (2007).
[CrossRef]

Huisken, J.

J. Huisken, J. Swoger, F. Del Bene, J. Wittbrodt, and E. H. K. Stelzer, Science 305, 1007 (2004).
[CrossRef]

Jahrling, N.

H. U. Dodt, U. Leischner, A. Schierloh, N. Jahrling, C. P. Mauch, K. Deininger, J. M. Deussing, M. Eder, W. Zieglgansberger, and K. Becker, Nat. Methods 4, 331 (2007).
[CrossRef]

Kak, A. C.

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

Konstantinides, N.

Krstajic, N.

N. Krstajic and S. J. Doran, Phys. Med. Biol. 51, 2055 (2006).
[CrossRef]

Laine, R.

J. McGinty, K. B. Tahir, R. Laine, C. B. Talbot, C. Dunsby, M. A. A. Neil, L. Quintana, J. Swoger, J. Sharpe, and P. M. W. French, J. Biophoton. 1, 390 (2008).
[CrossRef]

Lamb, J. R.

Leischner, U.

H. U. Dodt, U. Leischner, A. Schierloh, N. Jahrling, C. P. Mauch, K. Deininger, J. M. Deussing, M. Eder, W. Zieglgansberger, and K. Becker, Nat. Methods 4, 331 (2007).
[CrossRef]

Lorbeer, C.

Lorbeer, R.

Lorén, C. E.

T. Alanentalo, A. Asayesh, H. Morrison, C. E. Lorén, D. Holmberg, J. Sharpe, and U. Ahlgren, Nat. Methods 4, 31 (2007).
[CrossRef]

Mauch, C. P.

H. U. Dodt, U. Leischner, A. Schierloh, N. Jahrling, C. P. Mauch, K. Deininger, J. M. Deussing, M. Eder, W. Zieglgansberger, and K. Becker, Nat. Methods 4, 331 (2007).
[CrossRef]

McGinty, J.

L. Chen, J. McGinty, H. B. Taylor, L. Bugeon, J. R. Lamb, M. J. Dallman, and P. M. W. French, Opt. Express 20, 7323 (2012).
[CrossRef]

J. McGinty, K. B. Tahir, R. Laine, C. B. Talbot, C. Dunsby, M. A. A. Neil, L. Quintana, J. Swoger, J. Sharpe, and P. M. W. French, J. Biophoton. 1, 390 (2008).
[CrossRef]

Meyer, H.

Meyer, M. G.

Miao, Q.

Mione, M.

A. Bassi, L. Fieramonti, C. D’Andrea, M. Mione, and G. Valentini, J. Biomed. Opt. 16, 100502 (2011).
[CrossRef]

Morrison, H.

T. Alanentalo, A. Asayesh, H. Morrison, C. E. Lorén, D. Holmberg, J. Sharpe, and U. Ahlgren, Nat. Methods 4, 31 (2007).
[CrossRef]

Neil, M. A. A.

J. McGinty, K. B. Tahir, R. Laine, C. B. Talbot, C. Dunsby, M. A. A. Neil, L. Quintana, J. Swoger, J. Sharpe, and P. M. W. French, J. Biophoton. 1, 390 (2008).
[CrossRef]

Nelson, A. C.

Neumann, T.

Ojeda, D. F. R.

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]

Quintana, L.

J. McGinty, K. B. Tahir, R. Laine, C. B. Talbot, C. Dunsby, M. A. A. Neil, L. Quintana, J. Swoger, J. Sharpe, and P. M. W. French, J. Biophoton. 1, 390 (2008).
[CrossRef]

Rieckher, M.

Ripoll, J.

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]

Sarasa-Renedo, A.

Schierloh, A.

H. U. Dodt, U. Leischner, A. Schierloh, N. Jahrling, C. P. Mauch, K. Deininger, J. M. Deussing, M. Eder, W. Zieglgansberger, and K. Becker, Nat. Methods 4, 331 (2007).
[CrossRef]

Seibel, E. J.

Sharpe, J.

J. McGinty, K. B. Tahir, R. Laine, C. B. Talbot, C. Dunsby, M. A. A. Neil, L. Quintana, J. Swoger, J. Sharpe, and P. M. W. French, J. Biophoton. 1, 390 (2008).
[CrossRef]

T. Alanentalo, A. Asayesh, H. Morrison, C. E. Lorén, D. Holmberg, J. Sharpe, and U. Ahlgren, Nat. Methods 4, 31 (2007).
[CrossRef]

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

J. R. Walls, J. G. Sled, J. Sharpe, and R. M. Henkelman, Phys. Med. Biol. 50, 4645 (2005).
[CrossRef]

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

Slaney, M.

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

Sled, J. G.

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

J. R. Walls, J. G. Sled, J. Sharpe, and R. M. Henkelman, Phys. Med. Biol. 50, 4645 (2005).
[CrossRef]

Stelzer, E. H. K.

J. Huisken, J. Swoger, F. Del Bene, J. Wittbrodt, and E. H. K. Stelzer, Science 305, 1007 (2004).
[CrossRef]

Swoger, J.

J. McGinty, K. B. Tahir, R. Laine, C. B. Talbot, C. Dunsby, M. A. A. Neil, L. Quintana, J. Swoger, J. Sharpe, and P. M. W. French, J. Biophoton. 1, 390 (2008).
[CrossRef]

J. Huisken, J. Swoger, F. Del Bene, J. Wittbrodt, and E. H. K. Stelzer, Science 305, 1007 (2004).
[CrossRef]

Tahir, K. B.

J. McGinty, K. B. Tahir, R. Laine, C. B. Talbot, C. Dunsby, M. A. A. Neil, L. Quintana, J. Swoger, J. Sharpe, and P. M. W. French, J. Biophoton. 1, 390 (2008).
[CrossRef]

Talbot, C. B.

J. McGinty, K. B. Tahir, R. Laine, C. B. Talbot, C. Dunsby, M. A. A. Neil, L. Quintana, J. Swoger, J. Sharpe, and P. M. W. French, J. Biophoton. 1, 390 (2008).
[CrossRef]

Tavernarakis, N.

Taylor, H. B.

Valentini, G.

A. Bassi, L. Fieramonti, C. D’Andrea, M. Mione, and G. Valentini, J. Biomed. Opt. 16, 100502 (2011).
[CrossRef]

Walls, J. R.

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

J. R. Walls, J. G. Sled, J. Sharpe, and R. M. Henkelman, Phys. Med. Biol. 50, 4645 (2005).
[CrossRef]

Wittbrodt, J.

J. Huisken, J. Swoger, F. Del Bene, J. Wittbrodt, and E. H. K. Stelzer, Science 305, 1007 (2004).
[CrossRef]

Zieglgansberger, W.

H. U. Dodt, U. Leischner, A. Schierloh, N. Jahrling, C. P. Mauch, K. Deininger, J. M. Deussing, M. Eder, W. Zieglgansberger, and K. Becker, Nat. Methods 4, 331 (2007).
[CrossRef]

Biomed. Opt. Express (1)

Comput. Methods Programs Biomed. (1)

M. A. Haidekker, Comput. Methods Programs Biomed. 80, 225 (2005).
[CrossRef]

J. Biomed. Opt. (1)

A. Bassi, L. Fieramonti, C. D’Andrea, M. Mione, and G. Valentini, J. Biomed. Opt. 16, 100502 (2011).
[CrossRef]

J. Biophoton. (1)

J. McGinty, K. B. Tahir, R. Laine, C. B. Talbot, C. Dunsby, M. A. A. Neil, L. Quintana, J. Swoger, J. Sharpe, and P. M. W. French, J. Biophoton. 1, 390 (2008).
[CrossRef]

Nat. Methods (2)

T. Alanentalo, A. Asayesh, H. Morrison, C. E. Lorén, D. Holmberg, J. Sharpe, and U. Ahlgren, Nat. Methods 4, 31 (2007).
[CrossRef]

H. U. Dodt, U. Leischner, A. Schierloh, N. Jahrling, C. P. Mauch, K. Deininger, J. M. Deussing, M. Eder, W. Zieglgansberger, and K. Becker, Nat. Methods 4, 331 (2007).
[CrossRef]

Opt. Express (2)

Opt. Lett. (1)

Phys. Med. Biol. (3)

N. Krstajic and S. J. Doran, Phys. Med. Biol. 51, 2055 (2006).
[CrossRef]

J. R. Walls, J. G. Sled, J. Sharpe, and R. M. Henkelman, Phys. Med. Biol. 50, 4645 (2005).
[CrossRef]

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

Science (2)

J. Huisken, J. Swoger, F. Del Bene, J. Wittbrodt, and E. H. K. Stelzer, Science 305, 1007 (2004).
[CrossRef]

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

Other (1)

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

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Figures (4)

Fig. 1.
Fig. 1.

Schematic of (a) “standard” and (b) dual-axis OPT system setup (inset shows optimal DOFs and the FPs of two imaging systems). DOF, depth of field; EF, emission filter; AP, aperture; L1, 25 mm focal length lens; L2, 50 mm focal length lens; FP, focal plane; ϕ, sample diameter.

Fig. 2.
Fig. 2.

Reconstructed XY and XZ image slices of a bead acquired with standard OPT (a), (b) and dual-axis OPT (c), (d).

Fig. 3.
Fig. 3.

Line plots and Gaussian fits through the Y axis of the reconstructed bead in Fig. 2 for the dual-axis (D, red line) and standard (S, black lines) OPT systems with the latter showing relative (solid) and normalized (dashed) line intensity data.

Fig. 4.
Fig. 4.

Reconstructed (maximum intensity projection) images of the tail of a 54 dpf zebrafish acquired with (a) dual-axis and (b), (c) standard OPT plotted for comparison with normalized intensity (a), (b) and absolute intensity (a), (c) scales. (d) Intensity line profiles from (a), (b) as indicated, where the standard OPT intensity data have been multiplied by 2 for clarity.

Equations (1)

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

DOF=nbath(nλNA2+n·eMa·NA),

Metrics