Abstract

Mesoscopic 3D imaging has become a widely used optical imaging technique to visualize intact biological specimens. Selective plane illumination microscopy (SPIM) visualizes samples up to a centimeter in size with micrometer resolution by 3D data stitching but is limited to fluorescent contrast. Optical projection tomography (OPT) works with fluorescent and nonfluorescent contrasts, but its resolution is limited in large samples. We present a hybrid setup (OPTiSPIM) combining the advantages of each technique. The combination of fluorescent and nonfluorescent high-resolution 3D data into integrated datasets enables a more extensive representation of mesoscopic biological samples. The modular concept of the OPTiSPIM facilitates incorporation of the transmission OPT modality into already established light sheet based imaging setups.

© 2014 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. J. Huisken, J. Swoger, F. Del Bene, J. Wittbrodt, and E. H. Stelzer, Science 305, 1007 (2004).
    [CrossRef]
  3. J. R. Walls, J. G. Sled, J. Sharpe, and R. M. Henkelman, Phys. Med. Biol. 50, 4645 (2005).
    [CrossRef]
  4. C. Vinegoni, C. Pitsouli, D. Razansky, N. Perrimon, and V. Ntziachristos, Nat. Methods 5, 45 (2008).
    [CrossRef]
  5. L. Quintana and J. Sharpe, Cold Spring Harb. Protoc. 2011, 664 (2011).
  6. H. Hama, H. Kurokawa, H. Kawano, R. Ando, T. Shimogori, H. Noda, K. Fukami, A. Sakaue-Sawano, and A. Miyawaki, Nat. Neurosci. 14, 1481 (2011).
    [CrossRef]
  7. J. Huisken and D. Y. Stainier, Development 136, 1963 (2009).
    [CrossRef]
  8. V. Ntziachristos, Nat. Methods 7, 603 (2010).
    [CrossRef]
  9. P. A. Santi, J. Histochem Cytochem. 59, 129 (2011).
  10. M. Hargrave, J. Bowles, and P. Koopman, Methods Mol. Biol. 326, 103 (2006).
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  12. J. B. Pawley, Handbook of Biological Confocal Microscopy, 3rd ed. (Springer, 2006).
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    [CrossRef]
  14. K. Greger, J. Swoger, and E. H. Stelzer, Rev. Sci. Instrum. 78, 023705 (2007).
    [CrossRef]
  15. J. Huisken and D. Y. Stainier, Opt. Lett. 32, 2608 (2007).
    [CrossRef]
  16. P. J. Keller, A. D. Schmidt, J. Wittbrodt, and E. H. Stelzer, Science 322, 1065 (2008).
    [CrossRef]
  17. J. Sharpe, Annu. Rev. Biomed. Eng. 6, 209 (2004).
    [CrossRef]
  18. C. J. Engelbrecht and E. H. Stelzer, Opt. Lett. 31, 1477 (2006).
    [CrossRef]
  19. S. Preibisch, S. Saalfeld, and P. Tomancak, Bioinformatics 25, 1463 (2009).
    [CrossRef]
  20. J. Schindelin, I. Arganda-Carreras, E. Frise, V. Kaynig, M. Longair, T. Pietzsch, S. Preibisch, C. Rueden, S. Saalfeld, B. Schmid, J. Y. Tinevez, D. J. White, V. Hartenstein, K. Eliceiri, P. Tomancak, and A. Cardona, Nat. Methods 9, 676 (2012).
    [CrossRef]
  21. R. Sambasivan, S. Kuratani, and S. Tajbakhsh, Development 138, 2401 (2011).
    [CrossRef]
  22. J. Swoger, P. Verveer, K. Greger, J. Huisken, and E. H. Stelzer, Opt. Express 15, 8029 (2007).
    [CrossRef]
  23. J. R. Walls, J. G. Sled, J. Sharpe, and R. M. Henkelman, Phys. Med. Biol. 52, 2775 (2007).
    [CrossRef]

2013 (1)

E. J. Gualda, T. Vale, P. Almada, J. A. Feijo, G. G. Martins, and N. Moreno, Nat. Methods 10, 599 (2013).
[CrossRef]

2012 (1)

J. Schindelin, I. Arganda-Carreras, E. Frise, V. Kaynig, M. Longair, T. Pietzsch, S. Preibisch, C. Rueden, S. Saalfeld, B. Schmid, J. Y. Tinevez, D. J. White, V. Hartenstein, K. Eliceiri, P. Tomancak, and A. Cardona, Nat. Methods 9, 676 (2012).
[CrossRef]

2011 (5)

R. Sambasivan, S. Kuratani, and S. Tajbakhsh, Development 138, 2401 (2011).
[CrossRef]

L. Quintana and J. Sharpe, Cold Spring Harb. Protoc. 2011, 586 (2011).

L. Quintana and J. Sharpe, Cold Spring Harb. Protoc. 2011, 664 (2011).

H. Hama, H. Kurokawa, H. Kawano, R. Ando, T. Shimogori, H. Noda, K. Fukami, A. Sakaue-Sawano, and A. Miyawaki, Nat. Neurosci. 14, 1481 (2011).
[CrossRef]

P. A. Santi, J. Histochem Cytochem. 59, 129 (2011).

2010 (1)

V. Ntziachristos, Nat. Methods 7, 603 (2010).
[CrossRef]

2009 (2)

S. Preibisch, S. Saalfeld, and P. Tomancak, Bioinformatics 25, 1463 (2009).
[CrossRef]

J. Huisken and D. Y. Stainier, Development 136, 1963 (2009).
[CrossRef]

2008 (2)

C. Vinegoni, C. Pitsouli, D. Razansky, N. Perrimon, and V. Ntziachristos, Nat. Methods 5, 45 (2008).
[CrossRef]

P. J. Keller, A. D. Schmidt, J. Wittbrodt, and E. H. Stelzer, Science 322, 1065 (2008).
[CrossRef]

2007 (4)

K. Greger, J. Swoger, and E. H. Stelzer, Rev. Sci. Instrum. 78, 023705 (2007).
[CrossRef]

J. Huisken and D. Y. Stainier, Opt. Lett. 32, 2608 (2007).
[CrossRef]

J. Swoger, P. Verveer, K. Greger, J. Huisken, and E. H. Stelzer, Opt. Express 15, 8029 (2007).
[CrossRef]

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

2006 (2)

C. J. Engelbrecht and E. H. Stelzer, Opt. Lett. 31, 1477 (2006).
[CrossRef]

M. Hargrave, J. Bowles, and P. Koopman, Methods Mol. Biol. 326, 103 (2006).

2005 (1)

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

2004 (2)

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

J. Sharpe, Annu. Rev. Biomed. Eng. 6, 209 (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.

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

Almada, P.

E. J. Gualda, T. Vale, P. Almada, J. A. Feijo, G. G. Martins, and N. Moreno, Nat. Methods 10, 599 (2013).
[CrossRef]

Ando, R.

H. Hama, H. Kurokawa, H. Kawano, R. Ando, T. Shimogori, H. Noda, K. Fukami, A. Sakaue-Sawano, and A. Miyawaki, Nat. Neurosci. 14, 1481 (2011).
[CrossRef]

Arganda-Carreras, I.

J. Schindelin, I. Arganda-Carreras, E. Frise, V. Kaynig, M. Longair, T. Pietzsch, S. Preibisch, C. Rueden, S. Saalfeld, B. Schmid, J. Y. Tinevez, D. J. White, V. Hartenstein, K. Eliceiri, P. Tomancak, and A. Cardona, Nat. Methods 9, 676 (2012).
[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]

Bowles, J.

M. Hargrave, J. Bowles, and P. Koopman, Methods Mol. Biol. 326, 103 (2006).

Cardona, A.

J. Schindelin, I. Arganda-Carreras, E. Frise, V. Kaynig, M. Longair, T. Pietzsch, S. Preibisch, C. Rueden, S. Saalfeld, B. Schmid, J. Y. Tinevez, D. J. White, V. Hartenstein, K. Eliceiri, P. Tomancak, and A. Cardona, Nat. Methods 9, 676 (2012).
[CrossRef]

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]

Del Bene, F.

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

Eliceiri, K.

J. Schindelin, I. Arganda-Carreras, E. Frise, V. Kaynig, M. Longair, T. Pietzsch, S. Preibisch, C. Rueden, S. Saalfeld, B. Schmid, J. Y. Tinevez, D. J. White, V. Hartenstein, K. Eliceiri, P. Tomancak, and A. Cardona, Nat. Methods 9, 676 (2012).
[CrossRef]

Engelbrecht, C. J.

Feijo, J. A.

E. J. Gualda, T. Vale, P. Almada, J. A. Feijo, G. G. Martins, and N. Moreno, Nat. Methods 10, 599 (2013).
[CrossRef]

Frise, E.

J. Schindelin, I. Arganda-Carreras, E. Frise, V. Kaynig, M. Longair, T. Pietzsch, S. Preibisch, C. Rueden, S. Saalfeld, B. Schmid, J. Y. Tinevez, D. J. White, V. Hartenstein, K. Eliceiri, P. Tomancak, and A. Cardona, Nat. Methods 9, 676 (2012).
[CrossRef]

Fukami, K.

H. Hama, H. Kurokawa, H. Kawano, R. Ando, T. Shimogori, H. Noda, K. Fukami, A. Sakaue-Sawano, and A. Miyawaki, Nat. Neurosci. 14, 1481 (2011).
[CrossRef]

Greger, K.

K. Greger, J. Swoger, and E. H. Stelzer, Rev. Sci. Instrum. 78, 023705 (2007).
[CrossRef]

J. Swoger, P. Verveer, K. Greger, J. Huisken, and E. H. Stelzer, Opt. Express 15, 8029 (2007).
[CrossRef]

Gualda, E. J.

E. J. Gualda, T. Vale, P. Almada, J. A. Feijo, G. G. Martins, and N. Moreno, Nat. Methods 10, 599 (2013).
[CrossRef]

Hama, H.

H. Hama, H. Kurokawa, H. Kawano, R. Ando, T. Shimogori, H. Noda, K. Fukami, A. Sakaue-Sawano, and A. Miyawaki, Nat. Neurosci. 14, 1481 (2011).
[CrossRef]

Hargrave, M.

M. Hargrave, J. Bowles, and P. Koopman, Methods Mol. Biol. 326, 103 (2006).

Hartenstein, V.

J. Schindelin, I. Arganda-Carreras, E. Frise, V. Kaynig, M. Longair, T. Pietzsch, S. Preibisch, C. Rueden, S. Saalfeld, B. Schmid, J. Y. Tinevez, D. J. White, V. Hartenstein, K. Eliceiri, P. Tomancak, and A. Cardona, Nat. Methods 9, 676 (2012).
[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]

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]

Huisken, J.

J. Huisken and D. Y. Stainier, Development 136, 1963 (2009).
[CrossRef]

J. Swoger, P. Verveer, K. Greger, J. Huisken, and E. H. Stelzer, Opt. Express 15, 8029 (2007).
[CrossRef]

J. Huisken and D. Y. Stainier, Opt. Lett. 32, 2608 (2007).
[CrossRef]

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

Kawano, H.

H. Hama, H. Kurokawa, H. Kawano, R. Ando, T. Shimogori, H. Noda, K. Fukami, A. Sakaue-Sawano, and A. Miyawaki, Nat. Neurosci. 14, 1481 (2011).
[CrossRef]

Kaynig, V.

J. Schindelin, I. Arganda-Carreras, E. Frise, V. Kaynig, M. Longair, T. Pietzsch, S. Preibisch, C. Rueden, S. Saalfeld, B. Schmid, J. Y. Tinevez, D. J. White, V. Hartenstein, K. Eliceiri, P. Tomancak, and A. Cardona, Nat. Methods 9, 676 (2012).
[CrossRef]

Keller, P. J.

P. J. Keller, A. D. Schmidt, J. Wittbrodt, and E. H. Stelzer, Science 322, 1065 (2008).
[CrossRef]

Koopman, P.

M. Hargrave, J. Bowles, and P. Koopman, Methods Mol. Biol. 326, 103 (2006).

Kuratani, S.

R. Sambasivan, S. Kuratani, and S. Tajbakhsh, Development 138, 2401 (2011).
[CrossRef]

Kurokawa, H.

H. Hama, H. Kurokawa, H. Kawano, R. Ando, T. Shimogori, H. Noda, K. Fukami, A. Sakaue-Sawano, and A. Miyawaki, Nat. Neurosci. 14, 1481 (2011).
[CrossRef]

Longair, M.

J. Schindelin, I. Arganda-Carreras, E. Frise, V. Kaynig, M. Longair, T. Pietzsch, S. Preibisch, C. Rueden, S. Saalfeld, B. Schmid, J. Y. Tinevez, D. J. White, V. Hartenstein, K. Eliceiri, P. Tomancak, and A. Cardona, Nat. Methods 9, 676 (2012).
[CrossRef]

Martins, G. G.

E. J. Gualda, T. Vale, P. Almada, J. A. Feijo, G. G. Martins, and N. Moreno, Nat. Methods 10, 599 (2013).
[CrossRef]

Miyawaki, A.

H. Hama, H. Kurokawa, H. Kawano, R. Ando, T. Shimogori, H. Noda, K. Fukami, A. Sakaue-Sawano, and A. Miyawaki, Nat. Neurosci. 14, 1481 (2011).
[CrossRef]

Moreno, N.

E. J. Gualda, T. Vale, P. Almada, J. A. Feijo, G. G. Martins, and N. Moreno, Nat. Methods 10, 599 (2013).
[CrossRef]

Noda, H.

H. Hama, H. Kurokawa, H. Kawano, R. Ando, T. Shimogori, H. Noda, K. Fukami, A. Sakaue-Sawano, and A. Miyawaki, Nat. Neurosci. 14, 1481 (2011).
[CrossRef]

Ntziachristos, V.

V. Ntziachristos, Nat. Methods 7, 603 (2010).
[CrossRef]

C. Vinegoni, C. Pitsouli, D. Razansky, N. Perrimon, and V. Ntziachristos, Nat. Methods 5, 45 (2008).
[CrossRef]

Pawley, J. B.

J. B. Pawley, Handbook of Biological Confocal Microscopy, 3rd ed. (Springer, 2006).

Perrimon, N.

C. Vinegoni, C. Pitsouli, D. Razansky, N. Perrimon, and V. Ntziachristos, Nat. Methods 5, 45 (2008).
[CrossRef]

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]

Pietzsch, T.

J. Schindelin, I. Arganda-Carreras, E. Frise, V. Kaynig, M. Longair, T. Pietzsch, S. Preibisch, C. Rueden, S. Saalfeld, B. Schmid, J. Y. Tinevez, D. J. White, V. Hartenstein, K. Eliceiri, P. Tomancak, and A. Cardona, Nat. Methods 9, 676 (2012).
[CrossRef]

Pitsouli, C.

C. Vinegoni, C. Pitsouli, D. Razansky, N. Perrimon, and V. Ntziachristos, Nat. Methods 5, 45 (2008).
[CrossRef]

Preibisch, S.

J. Schindelin, I. Arganda-Carreras, E. Frise, V. Kaynig, M. Longair, T. Pietzsch, S. Preibisch, C. Rueden, S. Saalfeld, B. Schmid, J. Y. Tinevez, D. J. White, V. Hartenstein, K. Eliceiri, P. Tomancak, and A. Cardona, Nat. Methods 9, 676 (2012).
[CrossRef]

S. Preibisch, S. Saalfeld, and P. Tomancak, Bioinformatics 25, 1463 (2009).
[CrossRef]

Quintana, L.

L. Quintana and J. Sharpe, Cold Spring Harb. Protoc. 2011, 586 (2011).

L. Quintana and J. Sharpe, Cold Spring Harb. Protoc. 2011, 664 (2011).

Razansky, D.

C. Vinegoni, C. Pitsouli, D. Razansky, N. Perrimon, and V. Ntziachristos, Nat. Methods 5, 45 (2008).
[CrossRef]

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]

Rueden, C.

J. Schindelin, I. Arganda-Carreras, E. Frise, V. Kaynig, M. Longair, T. Pietzsch, S. Preibisch, C. Rueden, S. Saalfeld, B. Schmid, J. Y. Tinevez, D. J. White, V. Hartenstein, K. Eliceiri, P. Tomancak, and A. Cardona, Nat. Methods 9, 676 (2012).
[CrossRef]

Saalfeld, S.

J. Schindelin, I. Arganda-Carreras, E. Frise, V. Kaynig, M. Longair, T. Pietzsch, S. Preibisch, C. Rueden, S. Saalfeld, B. Schmid, J. Y. Tinevez, D. J. White, V. Hartenstein, K. Eliceiri, P. Tomancak, and A. Cardona, Nat. Methods 9, 676 (2012).
[CrossRef]

S. Preibisch, S. Saalfeld, and P. Tomancak, Bioinformatics 25, 1463 (2009).
[CrossRef]

Sakaue-Sawano, A.

H. Hama, H. Kurokawa, H. Kawano, R. Ando, T. Shimogori, H. Noda, K. Fukami, A. Sakaue-Sawano, and A. Miyawaki, Nat. Neurosci. 14, 1481 (2011).
[CrossRef]

Sambasivan, R.

R. Sambasivan, S. Kuratani, and S. Tajbakhsh, Development 138, 2401 (2011).
[CrossRef]

Santi, P. A.

P. A. Santi, J. Histochem Cytochem. 59, 129 (2011).

Schindelin, J.

J. Schindelin, I. Arganda-Carreras, E. Frise, V. Kaynig, M. Longair, T. Pietzsch, S. Preibisch, C. Rueden, S. Saalfeld, B. Schmid, J. Y. Tinevez, D. J. White, V. Hartenstein, K. Eliceiri, P. Tomancak, and A. Cardona, Nat. Methods 9, 676 (2012).
[CrossRef]

Schmid, B.

J. Schindelin, I. Arganda-Carreras, E. Frise, V. Kaynig, M. Longair, T. Pietzsch, S. Preibisch, C. Rueden, S. Saalfeld, B. Schmid, J. Y. Tinevez, D. J. White, V. Hartenstein, K. Eliceiri, P. Tomancak, and A. Cardona, Nat. Methods 9, 676 (2012).
[CrossRef]

Schmidt, A. D.

P. J. Keller, A. D. Schmidt, J. Wittbrodt, and E. H. Stelzer, Science 322, 1065 (2008).
[CrossRef]

Sharpe, J.

L. Quintana and J. Sharpe, Cold Spring Harb. Protoc. 2011, 586 (2011).

L. Quintana and J. Sharpe, Cold Spring Harb. Protoc. 2011, 664 (2011).

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, Annu. Rev. Biomed. Eng. 6, 209 (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]

Shimogori, T.

H. Hama, H. Kurokawa, H. Kawano, R. Ando, T. Shimogori, H. Noda, K. Fukami, A. Sakaue-Sawano, and A. Miyawaki, Nat. Neurosci. 14, 1481 (2011).
[CrossRef]

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]

Stainier, D. Y.

J. Huisken and D. Y. Stainier, Development 136, 1963 (2009).
[CrossRef]

J. Huisken and D. Y. Stainier, Opt. Lett. 32, 2608 (2007).
[CrossRef]

Stelzer, E. H.

P. J. Keller, A. D. Schmidt, J. Wittbrodt, and E. H. Stelzer, Science 322, 1065 (2008).
[CrossRef]

J. Swoger, P. Verveer, K. Greger, J. Huisken, and E. H. Stelzer, Opt. Express 15, 8029 (2007).
[CrossRef]

K. Greger, J. Swoger, and E. H. Stelzer, Rev. Sci. Instrum. 78, 023705 (2007).
[CrossRef]

C. J. Engelbrecht and E. H. Stelzer, Opt. Lett. 31, 1477 (2006).
[CrossRef]

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

Swoger, J.

K. Greger, J. Swoger, and E. H. Stelzer, Rev. Sci. Instrum. 78, 023705 (2007).
[CrossRef]

J. Swoger, P. Verveer, K. Greger, J. Huisken, and E. H. Stelzer, Opt. Express 15, 8029 (2007).
[CrossRef]

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

Tajbakhsh, S.

R. Sambasivan, S. Kuratani, and S. Tajbakhsh, Development 138, 2401 (2011).
[CrossRef]

Tinevez, J. Y.

J. Schindelin, I. Arganda-Carreras, E. Frise, V. Kaynig, M. Longair, T. Pietzsch, S. Preibisch, C. Rueden, S. Saalfeld, B. Schmid, J. Y. Tinevez, D. J. White, V. Hartenstein, K. Eliceiri, P. Tomancak, and A. Cardona, Nat. Methods 9, 676 (2012).
[CrossRef]

Tomancak, P.

J. Schindelin, I. Arganda-Carreras, E. Frise, V. Kaynig, M. Longair, T. Pietzsch, S. Preibisch, C. Rueden, S. Saalfeld, B. Schmid, J. Y. Tinevez, D. J. White, V. Hartenstein, K. Eliceiri, P. Tomancak, and A. Cardona, Nat. Methods 9, 676 (2012).
[CrossRef]

S. Preibisch, S. Saalfeld, and P. Tomancak, Bioinformatics 25, 1463 (2009).
[CrossRef]

Vale, T.

E. J. Gualda, T. Vale, P. Almada, J. A. Feijo, G. G. Martins, and N. Moreno, Nat. Methods 10, 599 (2013).
[CrossRef]

Verveer, P.

Vinegoni, C.

C. Vinegoni, C. Pitsouli, D. Razansky, N. Perrimon, and V. Ntziachristos, Nat. Methods 5, 45 (2008).
[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]

White, D. J.

J. Schindelin, I. Arganda-Carreras, E. Frise, V. Kaynig, M. Longair, T. Pietzsch, S. Preibisch, C. Rueden, S. Saalfeld, B. Schmid, J. Y. Tinevez, D. J. White, V. Hartenstein, K. Eliceiri, P. Tomancak, and A. Cardona, Nat. Methods 9, 676 (2012).
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Supplementary Material (1)

» Media 1: AVI (4011 KB)     

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Figures (3)

Fig. 1.
Fig. 1.

OPTiSPIM setup consists of four major modules: (1) SPIM illumination, (2) transmission illumination, (3) sample positioning, and (4) detection (including fluorescence OPT epiillumination). Module 1 forms the SPIM light sheet. AOTF, acousto-optic tunable filter; RSM, resonant scan mirror; SL, SPIM illumination lens; M, mirror; CL, cylindrical lens; HS, horizontal slit; VS, vertical slit; OL, objective lens. Module 2 trans-illuminates the sample in a Köhler configuration. TIL, transmission illumination lens; TI, transmission illumination iris. Module 3 manipulates the sample during the scan and supports it in the immersion medium. IMC, immersion medium chamber; S, sample. Module 4 images the sample onto the camera. DI, detection iris; EF, emission filter; TL, tube lens. OPT fluorescence excitation can be coupled in if needed. DM, dichroic mirror; FEX, fluorescence excitation lamp.

Fig. 2.
Fig. 2.

Fluorescent OPT scan compared to (fluorescent) SPIM scan, both imaged in the OPTiSPIM. The high endothelial venules (HEV) of a murine lymph node are visualized. (a), (d) full field of view, (b), (e) lateral resolution, (c), (f) axial resolution. Scale bar: 200 μm, scale bar inset: 50 μm.

Fig. 3.
Fig. 3.

(a) OPTiSPIM scan of an embryonic day 12.5 (E12.5) mouse head (maximum value projections), left: projection along the dorsoventral axis, right: along the lateral axis. Red: differentiating muscles, green: developing nerves (fluorescent SPIM scans). Blue: eye pigmentation (reconstructed transmission OPT scans). (b) Fluorescence OPT scan (Bioptonics 3001 scanner) compared with (c) SPIM scan of the dashed yellow-boxed region of interest. (d) Dorsoventral projection and (e) lateral projection of a region of interest around the eye [orange boxes in (a)]. The differentiating extraocular muscles (red) start to surround the eye (arrowheads). [See also (Media 1).] Scale bar: (a) 1000 μm, (b)–(e) 400 μm.

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