Abstract

The influence of semicrystalline order on the second-harmonic generation (SHG) efficiency in the anisotropic bands of Drosophila melanogaster sarcomeres from larval and adult muscle has been investigated. Differences in the semicrystalline order were obtained by using wild-type and mutant strains containing different amounts of headless myosin. The reduction in semicrystalline order without altering the chemical composition of myofibrils was achieved by observing highly stretched sarcomeres and by inducing a loss of viability in myocytes. In all cases the reduction of semicrystalline order in anisotropic bands of myocytes resulted in a substantial decrease in SHG. Second-harmonic imaging during periodic contractions of myocytes revealed higher intensities when sarcomeres were in the relaxed state compared with the contracted state. This study demonstrates that an ordered semicrystalline arrangement of anisotropic bands plays a determining role in the efficiency of SHG in myocytes.

© 2007 Optical Society of America

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    [CrossRef]

2006 (3)

S. V. Plotnikov, A. C. Millard, P. J. Campagnola, and W. A. Mohler, "Characterization of the myosin-based source for second-harmonic generation from muscle sarcomeres," Biophys. J. 90, 693-703 (2006).
[CrossRef]

C. Greenhalgh, B. Stewart, R. Cisek, N. Prent, A. Major, and V. Barzda, "Dynamic investigation of Drosophila myocytes with second-harmonic generations microscopy," in Biomedical Photonics, P. Mathieu, ed., Proc. SPIE 6343, 6343081-6343088 (2006).

A. Major, R. Cisek, C. Greenhalgh, N. Prent, and V. Barzda, "A diode-pumped high-power extended cavity femtosecond Yb:KGW laser: from development to applications in nonlinear microscopy," in Ultra-Short Pulse Optics: Lasers, Sources and Applications, P. Mathieu, ed., Proc. SPIE 6343, 6343451-6343458 (2006).

2005 (3)

C. Greenhalgh, V. Barzda, S. Elmore, J. van Beek, J. Aus der Au, and J. Squier, "Visualization of nanocontractions in cardiomyocytes by simultaneous detection of second- and third-harmonic generation and multiphoton excitation fluorescence microscopy," in Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XII, J.-A. Conchello, C. J. Cogswell, and T. Wilson, eds., Proc. SPIE 5701, 128-135 (2005).
[CrossRef]

C. K. Sun, "Higher harmonic generation microscopy," Adv. Biochem. Eng. Biotechnol. 95, 17-56 (2005).
[PubMed]

V. Barzda, C. Greenhalgh, J. Aus der Au, S. Elmore, J. H. G. M. Van Beek, and J. Squier, "Visualization of mitochondria in cardiomyocytes by simultaneous harmonic generation and fluorescence microscopy," Opt. Express 13, 8263-8276 (2005).
[CrossRef] [PubMed]

2004 (4)

T. Boulesteix, E. Beaurepaire, M. P. Sauviat, and M. C. Schanne-Klein, "Second-harmonic microscopy of unstained living cardiac myocytes: measurements of sarcomere length with 20-nm accuracy," Opt. Lett. 29, 2031-2033 (2004).
[CrossRef] [PubMed]

J. X. Cheng and X. S. Xie, "Coherent anti-Stokes Raman scattering microscopy: Instrumentation, theory, and applications," J. Phys. Chem. B 108, 827-840 (2004).
[CrossRef]

S. W. Chu, S. Y. Chen, G. W. Chern, T. H. Tsai, Y. C. Chen, B. L. Lin, and C. K. Sun, "Studies of x((2))/x((3)) tensors in submicron-scaled bio-tissues by polarization harmonics optical microscopy," Biophys. J. 86, 3914-3922 (2004).
[CrossRef] [PubMed]

M. Both, M. Vogel, O. Friedrich, F. von Wegner, T. Kunsting, R. H. A. Fink, and D. Uttenweiler, "Second-harmonic imaging of intrinsic signals in muscle fibers in situ," J. Biomed. Opt. 9, 882-892 (2004).
[CrossRef] [PubMed]

2003 (3)

A. C. Millard, P. J. Campagnola, W. Mohler, A. Lewis, and L. M. Loew, "Second harmonic imaging microscopy," Methods Enzymol. 361, 47-69 (2003).
[CrossRef] [PubMed]

W. R. Zipfel, R. M. Williams, and W. W. Webb, "Nonlinear magic: multiphoton microscopy in the biosciences," Nat. Biotechnol. 21, 1368-1376 (2003).
[CrossRef]

S. W. Chu, S. Y. Chen, T. H. Tsai, T. M. Liu, C. Y. Lin, H. J. Tsai, and C. K. Sun, "In vivo developmental biology study using noninvasive multiharmonic generation microscopy," Opt. Express 11, 3093-3099 (2003).
[CrossRef] [PubMed]

2002 (2)

P. J. Campagnola, A. C. Millard, M. Terasaki, P. E. Hoppe, C. J. Malone, and W. A. Mohler, "Three-dimensional high-resolution second-harmonic generation imaging of endogenous structural proteins in biological tissues," Biophys. J. 82, 493-508 (2002).
[CrossRef]

S. W. Chu, I. H. Chen, T. M. Liu, C. K. Sun, S. P. Lee, B. L. Lin, P. C. Cheng, M. X. Kuo, D. J. Lin, and H. L. Liu, "Nonlinear bio-photonic crystal effects revealed with multimodal nonlinear microscopy," J. Microsc. 208, 190-200 (2002).
[CrossRef] [PubMed]

2001 (2)

J. A. Squier and M. Muller, "High resolution nonlinear microscopy: A review of sources and methods for achieving optimal imaging," Rev. Sci. Instrum. 72, 2855-2867 (2001).
[CrossRef]

S. W. Chu, I. H. Chen, T. M. Liu, P. C. Chen, C. K. Sun, and B. L. Lin, "Multimodal nonlinear spectral microscopy based on a femtosecond Cr : forsterite laser," Opt. Lett. 26, 1909-1911 (2001).
[CrossRef]

2000 (2)

1999 (1)

R. M. Cripps, J. A. Suggs, and S. I. Bernstein, "Assembly of thick filaments and myofibrils occurs in the absence of the myosin head," EMBO J. 18, 1793-1804 (1999).
[CrossRef] [PubMed]

1998 (1)

M. Muller, J. Squier, K. R. Wilson, and G. J. Brakenhoff, "3D microscopy of transparent objects using third-harmonic generation," J. Microsc. 191, 266-274 (1998).
[CrossRef] [PubMed]

1997 (1)

1986 (1)

1962 (1)

J. A. Armstrong, N. Bloembergen, J. Ducuing, and P. S. Pershan, "Interactions between light waves in a nonlinear dielectric," Phys. Rev. 127, 1918-1939 (1962).
[CrossRef]

1953 (2)

H. E. Huxley, "Electron microscope studies of the organisation of the filaments in striated muscle," Biochim. Biophys. Acta 12, 387-400 (1953).
[CrossRef] [PubMed]

J. Hanson and H. E. Huxley, "Structural basis of the cross-striations in muscle," Nature 172, 530-532 (1953).
[CrossRef] [PubMed]

Alfano, R. R.

Armstrong, J. A.

J. A. Armstrong, N. Bloembergen, J. Ducuing, and P. S. Pershan, "Interactions between light waves in a nonlinear dielectric," Phys. Rev. 127, 1918-1939 (1962).
[CrossRef]

Barzda, V.

A. Major, R. Cisek, C. Greenhalgh, N. Prent, and V. Barzda, "A diode-pumped high-power extended cavity femtosecond Yb:KGW laser: from development to applications in nonlinear microscopy," in Ultra-Short Pulse Optics: Lasers, Sources and Applications, P. Mathieu, ed., Proc. SPIE 6343, 6343451-6343458 (2006).

C. Greenhalgh, B. Stewart, R. Cisek, N. Prent, A. Major, and V. Barzda, "Dynamic investigation of Drosophila myocytes with second-harmonic generations microscopy," in Biomedical Photonics, P. Mathieu, ed., Proc. SPIE 6343, 6343081-6343088 (2006).

C. Greenhalgh, V. Barzda, S. Elmore, J. van Beek, J. Aus der Au, and J. Squier, "Visualization of nanocontractions in cardiomyocytes by simultaneous detection of second- and third-harmonic generation and multiphoton excitation fluorescence microscopy," in Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XII, J.-A. Conchello, C. J. Cogswell, and T. Wilson, eds., Proc. SPIE 5701, 128-135 (2005).
[CrossRef]

V. Barzda, C. Greenhalgh, J. Aus der Au, S. Elmore, J. H. G. M. Van Beek, and J. Squier, "Visualization of mitochondria in cardiomyocytes by simultaneous harmonic generation and fluorescence microscopy," Opt. Express 13, 8263-8276 (2005).
[CrossRef] [PubMed]

N. Prent, R. Cisek, C. Greenhalgh, J. Aus der Au, J. Squier, and V. Barzda, "Imaging individual chloroplasts simultaneously with third- and second-harmonic generation and multiphoton excitation fluorescence microscopy," in Photosynthesis: Fundamental Aspects to Global Perspectives, A.Van der Est and D.Bruce, eds. (Allen Press, 2005), pp. 1037-1039.

C. Greenhalgh, R. Cisek, B. Stewart, and V. Barzda, "Dynamic and structural visualization of muscle structure in Drosophila with multimodal harmonic generation microscopy," in Biomedical Optics Topical Meeting of OSA Proceedings (Optical Society of America, 2006) (to be published).

V. Barzda, "Non-linear Contrast Mechanisms for Optical Microscopy," in Biophysical Techniques in Photosynthesis, T.Aartsma and J.Matysik, eds. (Springer, 2006), to be published.

Beaurepaire, E.

Bernstein, S. I.

R. M. Cripps, J. A. Suggs, and S. I. Bernstein, "Assembly of thick filaments and myofibrils occurs in the absence of the myosin head," EMBO J. 18, 1793-1804 (1999).
[CrossRef] [PubMed]

Blanchard-Desce, M.

Bloembergen, N.

J. A. Armstrong, N. Bloembergen, J. Ducuing, and P. S. Pershan, "Interactions between light waves in a nonlinear dielectric," Phys. Rev. 127, 1918-1939 (1962).
[CrossRef]

Both, M.

M. Both, M. Vogel, O. Friedrich, F. von Wegner, T. Kunsting, R. H. A. Fink, and D. Uttenweiler, "Second-harmonic imaging of intrinsic signals in muscle fibers in situ," J. Biomed. Opt. 9, 882-892 (2004).
[CrossRef] [PubMed]

Boulesteix, T.

Boyd, R. W.

R. W. Boyd, Nonlinear Optics, 2nd ed. (Academic, 2003).

Brakenhoff, G. J.

M. Muller, J. Squier, K. R. Wilson, and G. J. Brakenhoff, "3D microscopy of transparent objects using third-harmonic generation," J. Microsc. 191, 266-274 (1998).
[CrossRef] [PubMed]

Campagnola, P. J.

S. V. Plotnikov, A. C. Millard, P. J. Campagnola, and W. A. Mohler, "Characterization of the myosin-based source for second-harmonic generation from muscle sarcomeres," Biophys. J. 90, 693-703 (2006).
[CrossRef]

A. C. Millard, P. J. Campagnola, W. Mohler, A. Lewis, and L. M. Loew, "Second harmonic imaging microscopy," Methods Enzymol. 361, 47-69 (2003).
[CrossRef] [PubMed]

P. J. Campagnola, A. C. Millard, M. Terasaki, P. E. Hoppe, C. J. Malone, and W. A. Mohler, "Three-dimensional high-resolution second-harmonic generation imaging of endogenous structural proteins in biological tissues," Biophys. J. 82, 493-508 (2002).
[CrossRef]

Chen, I. H.

S. W. Chu, I. H. Chen, T. M. Liu, C. K. Sun, S. P. Lee, B. L. Lin, P. C. Cheng, M. X. Kuo, D. J. Lin, and H. L. Liu, "Nonlinear bio-photonic crystal effects revealed with multimodal nonlinear microscopy," J. Microsc. 208, 190-200 (2002).
[CrossRef] [PubMed]

S. W. Chu, I. H. Chen, T. M. Liu, P. C. Chen, C. K. Sun, and B. L. Lin, "Multimodal nonlinear spectral microscopy based on a femtosecond Cr : forsterite laser," Opt. Lett. 26, 1909-1911 (2001).
[CrossRef]

Chen, P. C.

Chen, S. Y.

S. W. Chu, S. Y. Chen, G. W. Chern, T. H. Tsai, Y. C. Chen, B. L. Lin, and C. K. Sun, "Studies of x((2))/x((3)) tensors in submicron-scaled bio-tissues by polarization harmonics optical microscopy," Biophys. J. 86, 3914-3922 (2004).
[CrossRef] [PubMed]

S. W. Chu, S. Y. Chen, T. H. Tsai, T. M. Liu, C. Y. Lin, H. J. Tsai, and C. K. Sun, "In vivo developmental biology study using noninvasive multiharmonic generation microscopy," Opt. Express 11, 3093-3099 (2003).
[CrossRef] [PubMed]

Chen, Y. C.

S. W. Chu, S. Y. Chen, G. W. Chern, T. H. Tsai, Y. C. Chen, B. L. Lin, and C. K. Sun, "Studies of x((2))/x((3)) tensors in submicron-scaled bio-tissues by polarization harmonics optical microscopy," Biophys. J. 86, 3914-3922 (2004).
[CrossRef] [PubMed]

Cheng, J. X.

J. X. Cheng and X. S. Xie, "Coherent anti-Stokes Raman scattering microscopy: Instrumentation, theory, and applications," J. Phys. Chem. B 108, 827-840 (2004).
[CrossRef]

Cheng, P. C.

S. W. Chu, I. H. Chen, T. M. Liu, C. K. Sun, S. P. Lee, B. L. Lin, P. C. Cheng, M. X. Kuo, D. J. Lin, and H. L. Liu, "Nonlinear bio-photonic crystal effects revealed with multimodal nonlinear microscopy," J. Microsc. 208, 190-200 (2002).
[CrossRef] [PubMed]

Chern, G. W.

S. W. Chu, S. Y. Chen, G. W. Chern, T. H. Tsai, Y. C. Chen, B. L. Lin, and C. K. Sun, "Studies of x((2))/x((3)) tensors in submicron-scaled bio-tissues by polarization harmonics optical microscopy," Biophys. J. 86, 3914-3922 (2004).
[CrossRef] [PubMed]

Chu, S. W.

S. W. Chu, S. Y. Chen, G. W. Chern, T. H. Tsai, Y. C. Chen, B. L. Lin, and C. K. Sun, "Studies of x((2))/x((3)) tensors in submicron-scaled bio-tissues by polarization harmonics optical microscopy," Biophys. J. 86, 3914-3922 (2004).
[CrossRef] [PubMed]

S. W. Chu, S. Y. Chen, T. H. Tsai, T. M. Liu, C. Y. Lin, H. J. Tsai, and C. K. Sun, "In vivo developmental biology study using noninvasive multiharmonic generation microscopy," Opt. Express 11, 3093-3099 (2003).
[CrossRef] [PubMed]

S. W. Chu, I. H. Chen, T. M. Liu, C. K. Sun, S. P. Lee, B. L. Lin, P. C. Cheng, M. X. Kuo, D. J. Lin, and H. L. Liu, "Nonlinear bio-photonic crystal effects revealed with multimodal nonlinear microscopy," J. Microsc. 208, 190-200 (2002).
[CrossRef] [PubMed]

S. W. Chu, I. H. Chen, T. M. Liu, P. C. Chen, C. K. Sun, and B. L. Lin, "Multimodal nonlinear spectral microscopy based on a femtosecond Cr : forsterite laser," Opt. Lett. 26, 1909-1911 (2001).
[CrossRef]

Cisek, R.

A. Major, R. Cisek, C. Greenhalgh, N. Prent, and V. Barzda, "A diode-pumped high-power extended cavity femtosecond Yb:KGW laser: from development to applications in nonlinear microscopy," in Ultra-Short Pulse Optics: Lasers, Sources and Applications, P. Mathieu, ed., Proc. SPIE 6343, 6343451-6343458 (2006).

C. Greenhalgh, B. Stewart, R. Cisek, N. Prent, A. Major, and V. Barzda, "Dynamic investigation of Drosophila myocytes with second-harmonic generations microscopy," in Biomedical Photonics, P. Mathieu, ed., Proc. SPIE 6343, 6343081-6343088 (2006).

N. Prent, R. Cisek, C. Greenhalgh, J. Aus der Au, J. Squier, and V. Barzda, "Imaging individual chloroplasts simultaneously with third- and second-harmonic generation and multiphoton excitation fluorescence microscopy," in Photosynthesis: Fundamental Aspects to Global Perspectives, A.Van der Est and D.Bruce, eds. (Allen Press, 2005), pp. 1037-1039.

C. Greenhalgh, R. Cisek, B. Stewart, and V. Barzda, "Dynamic and structural visualization of muscle structure in Drosophila with multimodal harmonic generation microscopy," in Biomedical Optics Topical Meeting of OSA Proceedings (Optical Society of America, 2006) (to be published).

Cripps, R. M.

R. M. Cripps, J. A. Suggs, and S. I. Bernstein, "Assembly of thick filaments and myofibrils occurs in the absence of the myosin head," EMBO J. 18, 1793-1804 (1999).
[CrossRef] [PubMed]

der Au, J. Aus

V. Barzda, C. Greenhalgh, J. Aus der Au, S. Elmore, J. H. G. M. Van Beek, and J. Squier, "Visualization of mitochondria in cardiomyocytes by simultaneous harmonic generation and fluorescence microscopy," Opt. Express 13, 8263-8276 (2005).
[CrossRef] [PubMed]

C. Greenhalgh, V. Barzda, S. Elmore, J. van Beek, J. Aus der Au, and J. Squier, "Visualization of nanocontractions in cardiomyocytes by simultaneous detection of second- and third-harmonic generation and multiphoton excitation fluorescence microscopy," in Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XII, J.-A. Conchello, C. J. Cogswell, and T. Wilson, eds., Proc. SPIE 5701, 128-135 (2005).
[CrossRef]

N. Prent, R. Cisek, C. Greenhalgh, J. Aus der Au, J. Squier, and V. Barzda, "Imaging individual chloroplasts simultaneously with third- and second-harmonic generation and multiphoton excitation fluorescence microscopy," in Photosynthesis: Fundamental Aspects to Global Perspectives, A.Van der Est and D.Bruce, eds. (Allen Press, 2005), pp. 1037-1039.

Deutsch, M.

Ducuing, J.

J. A. Armstrong, N. Bloembergen, J. Ducuing, and P. S. Pershan, "Interactions between light waves in a nonlinear dielectric," Phys. Rev. 127, 1918-1939 (1962).
[CrossRef]

Elmore, S.

V. Barzda, C. Greenhalgh, J. Aus der Au, S. Elmore, J. H. G. M. Van Beek, and J. Squier, "Visualization of mitochondria in cardiomyocytes by simultaneous harmonic generation and fluorescence microscopy," Opt. Express 13, 8263-8276 (2005).
[CrossRef] [PubMed]

C. Greenhalgh, V. Barzda, S. Elmore, J. van Beek, J. Aus der Au, and J. Squier, "Visualization of nanocontractions in cardiomyocytes by simultaneous detection of second- and third-harmonic generation and multiphoton excitation fluorescence microscopy," in Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XII, J.-A. Conchello, C. J. Cogswell, and T. Wilson, eds., Proc. SPIE 5701, 128-135 (2005).
[CrossRef]

Fink, R. H. A.

M. Both, M. Vogel, O. Friedrich, F. von Wegner, T. Kunsting, R. H. A. Fink, and D. Uttenweiler, "Second-harmonic imaging of intrinsic signals in muscle fibers in situ," J. Biomed. Opt. 9, 882-892 (2004).
[CrossRef] [PubMed]

Freund, I.

Friedrich, O.

M. Both, M. Vogel, O. Friedrich, F. von Wegner, T. Kunsting, R. H. A. Fink, and D. Uttenweiler, "Second-harmonic imaging of intrinsic signals in muscle fibers in situ," J. Biomed. Opt. 9, 882-892 (2004).
[CrossRef] [PubMed]

Greenhalgh, C.

A. Major, R. Cisek, C. Greenhalgh, N. Prent, and V. Barzda, "A diode-pumped high-power extended cavity femtosecond Yb:KGW laser: from development to applications in nonlinear microscopy," in Ultra-Short Pulse Optics: Lasers, Sources and Applications, P. Mathieu, ed., Proc. SPIE 6343, 6343451-6343458 (2006).

C. Greenhalgh, B. Stewart, R. Cisek, N. Prent, A. Major, and V. Barzda, "Dynamic investigation of Drosophila myocytes with second-harmonic generations microscopy," in Biomedical Photonics, P. Mathieu, ed., Proc. SPIE 6343, 6343081-6343088 (2006).

C. Greenhalgh, V. Barzda, S. Elmore, J. van Beek, J. Aus der Au, and J. Squier, "Visualization of nanocontractions in cardiomyocytes by simultaneous detection of second- and third-harmonic generation and multiphoton excitation fluorescence microscopy," in Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XII, J.-A. Conchello, C. J. Cogswell, and T. Wilson, eds., Proc. SPIE 5701, 128-135 (2005).
[CrossRef]

V. Barzda, C. Greenhalgh, J. Aus der Au, S. Elmore, J. H. G. M. Van Beek, and J. Squier, "Visualization of mitochondria in cardiomyocytes by simultaneous harmonic generation and fluorescence microscopy," Opt. Express 13, 8263-8276 (2005).
[CrossRef] [PubMed]

N. Prent, R. Cisek, C. Greenhalgh, J. Aus der Au, J. Squier, and V. Barzda, "Imaging individual chloroplasts simultaneously with third- and second-harmonic generation and multiphoton excitation fluorescence microscopy," in Photosynthesis: Fundamental Aspects to Global Perspectives, A.Van der Est and D.Bruce, eds. (Allen Press, 2005), pp. 1037-1039.

C. Greenhalgh, R. Cisek, B. Stewart, and V. Barzda, "Dynamic and structural visualization of muscle structure in Drosophila with multimodal harmonic generation microscopy," in Biomedical Optics Topical Meeting of OSA Proceedings (Optical Society of America, 2006) (to be published).

Guo, Y. C.

Hanson, J.

J. Hanson and H. E. Huxley, "Structural basis of the cross-striations in muscle," Nature 172, 530-532 (1953).
[CrossRef] [PubMed]

Harris, D.

Ho, P. P.

Hoppe, P. E.

P. J. Campagnola, A. C. Millard, M. Terasaki, P. E. Hoppe, C. J. Malone, and W. A. Mohler, "Three-dimensional high-resolution second-harmonic generation imaging of endogenous structural proteins in biological tissues," Biophys. J. 82, 493-508 (2002).
[CrossRef]

Huxley, H. E.

J. Hanson and H. E. Huxley, "Structural basis of the cross-striations in muscle," Nature 172, 530-532 (1953).
[CrossRef] [PubMed]

H. E. Huxley, "Electron microscope studies of the organisation of the filaments in striated muscle," Biochim. Biophys. Acta 12, 387-400 (1953).
[CrossRef] [PubMed]

König, K.

K. König, "Multiphoton microscopy in life sciences," J. Microsc. 200, 83-104 (2000).
[CrossRef] [PubMed]

Kunsting, T.

M. Both, M. Vogel, O. Friedrich, F. von Wegner, T. Kunsting, R. H. A. Fink, and D. Uttenweiler, "Second-harmonic imaging of intrinsic signals in muscle fibers in situ," J. Biomed. Opt. 9, 882-892 (2004).
[CrossRef] [PubMed]

Kuo, M. X.

S. W. Chu, I. H. Chen, T. M. Liu, C. K. Sun, S. P. Lee, B. L. Lin, P. C. Cheng, M. X. Kuo, D. J. Lin, and H. L. Liu, "Nonlinear bio-photonic crystal effects revealed with multimodal nonlinear microscopy," J. Microsc. 208, 190-200 (2002).
[CrossRef] [PubMed]

Lee, S. P.

S. W. Chu, I. H. Chen, T. M. Liu, C. K. Sun, S. P. Lee, B. L. Lin, P. C. Cheng, M. X. Kuo, D. J. Lin, and H. L. Liu, "Nonlinear bio-photonic crystal effects revealed with multimodal nonlinear microscopy," J. Microsc. 208, 190-200 (2002).
[CrossRef] [PubMed]

Lewis, A.

A. C. Millard, P. J. Campagnola, W. Mohler, A. Lewis, and L. M. Loew, "Second harmonic imaging microscopy," Methods Enzymol. 361, 47-69 (2003).
[CrossRef] [PubMed]

Lin, B. L.

S. W. Chu, S. Y. Chen, G. W. Chern, T. H. Tsai, Y. C. Chen, B. L. Lin, and C. K. Sun, "Studies of x((2))/x((3)) tensors in submicron-scaled bio-tissues by polarization harmonics optical microscopy," Biophys. J. 86, 3914-3922 (2004).
[CrossRef] [PubMed]

S. W. Chu, I. H. Chen, T. M. Liu, C. K. Sun, S. P. Lee, B. L. Lin, P. C. Cheng, M. X. Kuo, D. J. Lin, and H. L. Liu, "Nonlinear bio-photonic crystal effects revealed with multimodal nonlinear microscopy," J. Microsc. 208, 190-200 (2002).
[CrossRef] [PubMed]

S. W. Chu, I. H. Chen, T. M. Liu, P. C. Chen, C. K. Sun, and B. L. Lin, "Multimodal nonlinear spectral microscopy based on a femtosecond Cr : forsterite laser," Opt. Lett. 26, 1909-1911 (2001).
[CrossRef]

Lin, C. Y.

Lin, D. J.

S. W. Chu, I. H. Chen, T. M. Liu, C. K. Sun, S. P. Lee, B. L. Lin, P. C. Cheng, M. X. Kuo, D. J. Lin, and H. L. Liu, "Nonlinear bio-photonic crystal effects revealed with multimodal nonlinear microscopy," J. Microsc. 208, 190-200 (2002).
[CrossRef] [PubMed]

Liu, F.

Liu, H. L.

S. W. Chu, I. H. Chen, T. M. Liu, C. K. Sun, S. P. Lee, B. L. Lin, P. C. Cheng, M. X. Kuo, D. J. Lin, and H. L. Liu, "Nonlinear bio-photonic crystal effects revealed with multimodal nonlinear microscopy," J. Microsc. 208, 190-200 (2002).
[CrossRef] [PubMed]

Liu, T. M.

Loew, L. M.

A. C. Millard, P. J. Campagnola, W. Mohler, A. Lewis, and L. M. Loew, "Second harmonic imaging microscopy," Methods Enzymol. 361, 47-69 (2003).
[CrossRef] [PubMed]

Major, A.

C. Greenhalgh, B. Stewart, R. Cisek, N. Prent, A. Major, and V. Barzda, "Dynamic investigation of Drosophila myocytes with second-harmonic generations microscopy," in Biomedical Photonics, P. Mathieu, ed., Proc. SPIE 6343, 6343081-6343088 (2006).

A. Major, R. Cisek, C. Greenhalgh, N. Prent, and V. Barzda, "A diode-pumped high-power extended cavity femtosecond Yb:KGW laser: from development to applications in nonlinear microscopy," in Ultra-Short Pulse Optics: Lasers, Sources and Applications, P. Mathieu, ed., Proc. SPIE 6343, 6343451-6343458 (2006).

Malone, C. J.

P. J. Campagnola, A. C. Millard, M. Terasaki, P. E. Hoppe, C. J. Malone, and W. A. Mohler, "Three-dimensional high-resolution second-harmonic generation imaging of endogenous structural proteins in biological tissues," Biophys. J. 82, 493-508 (2002).
[CrossRef]

Mertz, J.

Millard, A. C.

S. V. Plotnikov, A. C. Millard, P. J. Campagnola, and W. A. Mohler, "Characterization of the myosin-based source for second-harmonic generation from muscle sarcomeres," Biophys. J. 90, 693-703 (2006).
[CrossRef]

A. C. Millard, P. J. Campagnola, W. Mohler, A. Lewis, and L. M. Loew, "Second harmonic imaging microscopy," Methods Enzymol. 361, 47-69 (2003).
[CrossRef] [PubMed]

P. J. Campagnola, A. C. Millard, M. Terasaki, P. E. Hoppe, C. J. Malone, and W. A. Mohler, "Three-dimensional high-resolution second-harmonic generation imaging of endogenous structural proteins in biological tissues," Biophys. J. 82, 493-508 (2002).
[CrossRef]

Mohler, W.

A. C. Millard, P. J. Campagnola, W. Mohler, A. Lewis, and L. M. Loew, "Second harmonic imaging microscopy," Methods Enzymol. 361, 47-69 (2003).
[CrossRef] [PubMed]

Mohler, W. A.

S. V. Plotnikov, A. C. Millard, P. J. Campagnola, and W. A. Mohler, "Characterization of the myosin-based source for second-harmonic generation from muscle sarcomeres," Biophys. J. 90, 693-703 (2006).
[CrossRef]

P. J. Campagnola, A. C. Millard, M. Terasaki, P. E. Hoppe, C. J. Malone, and W. A. Mohler, "Three-dimensional high-resolution second-harmonic generation imaging of endogenous structural proteins in biological tissues," Biophys. J. 82, 493-508 (2002).
[CrossRef]

Moreaux, L.

Muller, M.

J. A. Squier and M. Muller, "High resolution nonlinear microscopy: A review of sources and methods for achieving optimal imaging," Rev. Sci. Instrum. 72, 2855-2867 (2001).
[CrossRef]

M. Muller, J. Squier, K. R. Wilson, and G. J. Brakenhoff, "3D microscopy of transparent objects using third-harmonic generation," J. Microsc. 191, 266-274 (1998).
[CrossRef] [PubMed]

Pershan, P. S.

J. A. Armstrong, N. Bloembergen, J. Ducuing, and P. S. Pershan, "Interactions between light waves in a nonlinear dielectric," Phys. Rev. 127, 1918-1939 (1962).
[CrossRef]

Plotnikov, S. V.

S. V. Plotnikov, A. C. Millard, P. J. Campagnola, and W. A. Mohler, "Characterization of the myosin-based source for second-harmonic generation from muscle sarcomeres," Biophys. J. 90, 693-703 (2006).
[CrossRef]

Prent, N.

A. Major, R. Cisek, C. Greenhalgh, N. Prent, and V. Barzda, "A diode-pumped high-power extended cavity femtosecond Yb:KGW laser: from development to applications in nonlinear microscopy," in Ultra-Short Pulse Optics: Lasers, Sources and Applications, P. Mathieu, ed., Proc. SPIE 6343, 6343451-6343458 (2006).

C. Greenhalgh, B. Stewart, R. Cisek, N. Prent, A. Major, and V. Barzda, "Dynamic investigation of Drosophila myocytes with second-harmonic generations microscopy," in Biomedical Photonics, P. Mathieu, ed., Proc. SPIE 6343, 6343081-6343088 (2006).

N. Prent, R. Cisek, C. Greenhalgh, J. Aus der Au, J. Squier, and V. Barzda, "Imaging individual chloroplasts simultaneously with third- and second-harmonic generation and multiphoton excitation fluorescence microscopy," in Photosynthesis: Fundamental Aspects to Global Perspectives, A.Van der Est and D.Bruce, eds. (Allen Press, 2005), pp. 1037-1039.

Sacks, P.

Sandre, O.

Sauviat, M. P.

Savage, H.

Schanne-Klein, M. C.

Schantz, S.

Squier, J.

C. Greenhalgh, V. Barzda, S. Elmore, J. van Beek, J. Aus der Au, and J. Squier, "Visualization of nanocontractions in cardiomyocytes by simultaneous detection of second- and third-harmonic generation and multiphoton excitation fluorescence microscopy," in Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XII, J.-A. Conchello, C. J. Cogswell, and T. Wilson, eds., Proc. SPIE 5701, 128-135 (2005).
[CrossRef]

V. Barzda, C. Greenhalgh, J. Aus der Au, S. Elmore, J. H. G. M. Van Beek, and J. Squier, "Visualization of mitochondria in cardiomyocytes by simultaneous harmonic generation and fluorescence microscopy," Opt. Express 13, 8263-8276 (2005).
[CrossRef] [PubMed]

M. Muller, J. Squier, K. R. Wilson, and G. J. Brakenhoff, "3D microscopy of transparent objects using third-harmonic generation," J. Microsc. 191, 266-274 (1998).
[CrossRef] [PubMed]

N. Prent, R. Cisek, C. Greenhalgh, J. Aus der Au, J. Squier, and V. Barzda, "Imaging individual chloroplasts simultaneously with third- and second-harmonic generation and multiphoton excitation fluorescence microscopy," in Photosynthesis: Fundamental Aspects to Global Perspectives, A.Van der Est and D.Bruce, eds. (Allen Press, 2005), pp. 1037-1039.

Squier, J. A.

J. A. Squier and M. Muller, "High resolution nonlinear microscopy: A review of sources and methods for achieving optimal imaging," Rev. Sci. Instrum. 72, 2855-2867 (2001).
[CrossRef]

Stewart, B.

C. Greenhalgh, B. Stewart, R. Cisek, N. Prent, A. Major, and V. Barzda, "Dynamic investigation of Drosophila myocytes with second-harmonic generations microscopy," in Biomedical Photonics, P. Mathieu, ed., Proc. SPIE 6343, 6343081-6343088 (2006).

C. Greenhalgh, R. Cisek, B. Stewart, and V. Barzda, "Dynamic and structural visualization of muscle structure in Drosophila with multimodal harmonic generation microscopy," in Biomedical Optics Topical Meeting of OSA Proceedings (Optical Society of America, 2006) (to be published).

Suggs, J. A.

R. M. Cripps, J. A. Suggs, and S. I. Bernstein, "Assembly of thick filaments and myofibrils occurs in the absence of the myosin head," EMBO J. 18, 1793-1804 (1999).
[CrossRef] [PubMed]

Sun, C. K.

C. K. Sun, "Higher harmonic generation microscopy," Adv. Biochem. Eng. Biotechnol. 95, 17-56 (2005).
[PubMed]

S. W. Chu, S. Y. Chen, G. W. Chern, T. H. Tsai, Y. C. Chen, B. L. Lin, and C. K. Sun, "Studies of x((2))/x((3)) tensors in submicron-scaled bio-tissues by polarization harmonics optical microscopy," Biophys. J. 86, 3914-3922 (2004).
[CrossRef] [PubMed]

S. W. Chu, S. Y. Chen, T. H. Tsai, T. M. Liu, C. Y. Lin, H. J. Tsai, and C. K. Sun, "In vivo developmental biology study using noninvasive multiharmonic generation microscopy," Opt. Express 11, 3093-3099 (2003).
[CrossRef] [PubMed]

S. W. Chu, I. H. Chen, T. M. Liu, C. K. Sun, S. P. Lee, B. L. Lin, P. C. Cheng, M. X. Kuo, D. J. Lin, and H. L. Liu, "Nonlinear bio-photonic crystal effects revealed with multimodal nonlinear microscopy," J. Microsc. 208, 190-200 (2002).
[CrossRef] [PubMed]

S. W. Chu, I. H. Chen, T. M. Liu, P. C. Chen, C. K. Sun, and B. L. Lin, "Multimodal nonlinear spectral microscopy based on a femtosecond Cr : forsterite laser," Opt. Lett. 26, 1909-1911 (2001).
[CrossRef]

Terasaki, M.

P. J. Campagnola, A. C. Millard, M. Terasaki, P. E. Hoppe, C. J. Malone, and W. A. Mohler, "Three-dimensional high-resolution second-harmonic generation imaging of endogenous structural proteins in biological tissues," Biophys. J. 82, 493-508 (2002).
[CrossRef]

Tsai, H. J.

Tsai, T. H.

S. W. Chu, S. Y. Chen, G. W. Chern, T. H. Tsai, Y. C. Chen, B. L. Lin, and C. K. Sun, "Studies of x((2))/x((3)) tensors in submicron-scaled bio-tissues by polarization harmonics optical microscopy," Biophys. J. 86, 3914-3922 (2004).
[CrossRef] [PubMed]

S. W. Chu, S. Y. Chen, T. H. Tsai, T. M. Liu, C. Y. Lin, H. J. Tsai, and C. K. Sun, "In vivo developmental biology study using noninvasive multiharmonic generation microscopy," Opt. Express 11, 3093-3099 (2003).
[CrossRef] [PubMed]

Uttenweiler, D.

M. Both, M. Vogel, O. Friedrich, F. von Wegner, T. Kunsting, R. H. A. Fink, and D. Uttenweiler, "Second-harmonic imaging of intrinsic signals in muscle fibers in situ," J. Biomed. Opt. 9, 882-892 (2004).
[CrossRef] [PubMed]

van Beek, J.

C. Greenhalgh, V. Barzda, S. Elmore, J. van Beek, J. Aus der Au, and J. Squier, "Visualization of nanocontractions in cardiomyocytes by simultaneous detection of second- and third-harmonic generation and multiphoton excitation fluorescence microscopy," in Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XII, J.-A. Conchello, C. J. Cogswell, and T. Wilson, eds., Proc. SPIE 5701, 128-135 (2005).
[CrossRef]

Van Beek, J. H. G. M.

Vogel, M.

M. Both, M. Vogel, O. Friedrich, F. von Wegner, T. Kunsting, R. H. A. Fink, and D. Uttenweiler, "Second-harmonic imaging of intrinsic signals in muscle fibers in situ," J. Biomed. Opt. 9, 882-892 (2004).
[CrossRef] [PubMed]

von Wegner, F.

M. Both, M. Vogel, O. Friedrich, F. von Wegner, T. Kunsting, R. H. A. Fink, and D. Uttenweiler, "Second-harmonic imaging of intrinsic signals in muscle fibers in situ," J. Biomed. Opt. 9, 882-892 (2004).
[CrossRef] [PubMed]

Webb, W. W.

W. R. Zipfel, R. M. Williams, and W. W. Webb, "Nonlinear magic: multiphoton microscopy in the biosciences," Nat. Biotechnol. 21, 1368-1376 (2003).
[CrossRef]

Williams, R. M.

W. R. Zipfel, R. M. Williams, and W. W. Webb, "Nonlinear magic: multiphoton microscopy in the biosciences," Nat. Biotechnol. 21, 1368-1376 (2003).
[CrossRef]

Wilson, K. R.

M. Muller, J. Squier, K. R. Wilson, and G. J. Brakenhoff, "3D microscopy of transparent objects using third-harmonic generation," J. Microsc. 191, 266-274 (1998).
[CrossRef] [PubMed]

Xie, X. S.

J. X. Cheng and X. S. Xie, "Coherent anti-Stokes Raman scattering microscopy: Instrumentation, theory, and applications," J. Phys. Chem. B 108, 827-840 (2004).
[CrossRef]

Zhadin, N.

Zipfel, W. R.

W. R. Zipfel, R. M. Williams, and W. W. Webb, "Nonlinear magic: multiphoton microscopy in the biosciences," Nat. Biotechnol. 21, 1368-1376 (2003).
[CrossRef]

Adv. Biochem. Eng. Biotechnol. (1)

C. K. Sun, "Higher harmonic generation microscopy," Adv. Biochem. Eng. Biotechnol. 95, 17-56 (2005).
[PubMed]

Biochim. Biophys. Acta (1)

H. E. Huxley, "Electron microscope studies of the organisation of the filaments in striated muscle," Biochim. Biophys. Acta 12, 387-400 (1953).
[CrossRef] [PubMed]

Biophys. J. (3)

S. V. Plotnikov, A. C. Millard, P. J. Campagnola, and W. A. Mohler, "Characterization of the myosin-based source for second-harmonic generation from muscle sarcomeres," Biophys. J. 90, 693-703 (2006).
[CrossRef]

S. W. Chu, S. Y. Chen, G. W. Chern, T. H. Tsai, Y. C. Chen, B. L. Lin, and C. K. Sun, "Studies of x((2))/x((3)) tensors in submicron-scaled bio-tissues by polarization harmonics optical microscopy," Biophys. J. 86, 3914-3922 (2004).
[CrossRef] [PubMed]

P. J. Campagnola, A. C. Millard, M. Terasaki, P. E. Hoppe, C. J. Malone, and W. A. Mohler, "Three-dimensional high-resolution second-harmonic generation imaging of endogenous structural proteins in biological tissues," Biophys. J. 82, 493-508 (2002).
[CrossRef]

EMBO J. (1)

R. M. Cripps, J. A. Suggs, and S. I. Bernstein, "Assembly of thick filaments and myofibrils occurs in the absence of the myosin head," EMBO J. 18, 1793-1804 (1999).
[CrossRef] [PubMed]

J. Biomed. Opt. (1)

M. Both, M. Vogel, O. Friedrich, F. von Wegner, T. Kunsting, R. H. A. Fink, and D. Uttenweiler, "Second-harmonic imaging of intrinsic signals in muscle fibers in situ," J. Biomed. Opt. 9, 882-892 (2004).
[CrossRef] [PubMed]

J. Microsc. (3)

M. Muller, J. Squier, K. R. Wilson, and G. J. Brakenhoff, "3D microscopy of transparent objects using third-harmonic generation," J. Microsc. 191, 266-274 (1998).
[CrossRef] [PubMed]

K. König, "Multiphoton microscopy in life sciences," J. Microsc. 200, 83-104 (2000).
[CrossRef] [PubMed]

S. W. Chu, I. H. Chen, T. M. Liu, C. K. Sun, S. P. Lee, B. L. Lin, P. C. Cheng, M. X. Kuo, D. J. Lin, and H. L. Liu, "Nonlinear bio-photonic crystal effects revealed with multimodal nonlinear microscopy," J. Microsc. 208, 190-200 (2002).
[CrossRef] [PubMed]

J. Phys. Chem. B (1)

J. X. Cheng and X. S. Xie, "Coherent anti-Stokes Raman scattering microscopy: Instrumentation, theory, and applications," J. Phys. Chem. B 108, 827-840 (2004).
[CrossRef]

Methods Enzymol. (1)

A. C. Millard, P. J. Campagnola, W. Mohler, A. Lewis, and L. M. Loew, "Second harmonic imaging microscopy," Methods Enzymol. 361, 47-69 (2003).
[CrossRef] [PubMed]

Nat. Biotechnol. (1)

W. R. Zipfel, R. M. Williams, and W. W. Webb, "Nonlinear magic: multiphoton microscopy in the biosciences," Nat. Biotechnol. 21, 1368-1376 (2003).
[CrossRef]

Nature (1)

J. Hanson and H. E. Huxley, "Structural basis of the cross-striations in muscle," Nature 172, 530-532 (1953).
[CrossRef] [PubMed]

Opt. Express (2)

Opt. Lett. (5)

Phys. Rev. (1)

J. A. Armstrong, N. Bloembergen, J. Ducuing, and P. S. Pershan, "Interactions between light waves in a nonlinear dielectric," Phys. Rev. 127, 1918-1939 (1962).
[CrossRef]

Proc. SPIE (3)

A. Major, R. Cisek, C. Greenhalgh, N. Prent, and V. Barzda, "A diode-pumped high-power extended cavity femtosecond Yb:KGW laser: from development to applications in nonlinear microscopy," in Ultra-Short Pulse Optics: Lasers, Sources and Applications, P. Mathieu, ed., Proc. SPIE 6343, 6343451-6343458 (2006).

C. Greenhalgh, B. Stewart, R. Cisek, N. Prent, A. Major, and V. Barzda, "Dynamic investigation of Drosophila myocytes with second-harmonic generations microscopy," in Biomedical Photonics, P. Mathieu, ed., Proc. SPIE 6343, 6343081-6343088 (2006).

C. Greenhalgh, V. Barzda, S. Elmore, J. van Beek, J. Aus der Au, and J. Squier, "Visualization of nanocontractions in cardiomyocytes by simultaneous detection of second- and third-harmonic generation and multiphoton excitation fluorescence microscopy," in Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XII, J.-A. Conchello, C. J. Cogswell, and T. Wilson, eds., Proc. SPIE 5701, 128-135 (2005).
[CrossRef]

Rev. Sci. Instrum. (1)

J. A. Squier and M. Muller, "High resolution nonlinear microscopy: A review of sources and methods for achieving optimal imaging," Rev. Sci. Instrum. 72, 2855-2867 (2001).
[CrossRef]

Other (4)

C. Greenhalgh, R. Cisek, B. Stewart, and V. Barzda, "Dynamic and structural visualization of muscle structure in Drosophila with multimodal harmonic generation microscopy," in Biomedical Optics Topical Meeting of OSA Proceedings (Optical Society of America, 2006) (to be published).

V. Barzda, "Non-linear Contrast Mechanisms for Optical Microscopy," in Biophysical Techniques in Photosynthesis, T.Aartsma and J.Matysik, eds. (Springer, 2006), to be published.

R. W. Boyd, Nonlinear Optics, 2nd ed. (Academic, 2003).

N. Prent, R. Cisek, C. Greenhalgh, J. Aus der Au, J. Squier, and V. Barzda, "Imaging individual chloroplasts simultaneously with third- and second-harmonic generation and multiphoton excitation fluorescence microscopy," in Photosynthesis: Fundamental Aspects to Global Perspectives, A.Van der Est and D.Bruce, eds. (Allen Press, 2005), pp. 1037-1039.

Supplementary Material (4)

» Media 1: AVI (495 KB)     
» Media 2: AVI (822 KB)     
» Media 3: AVI (587 KB)     
» Media 4: AVI (274 KB)     

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Figures (8)

Fig. 1
Fig. 1

Structural schematic of two sarcomeres. The main actin (thin) and myosin (thick) filaments are shown. The anisotropic band (A-band) is the region where myosin is located, while the isotropic region (I-band) is occupied by actin and titin. The small bare region in the center of the myosin, where there are no myosin heads, is called the H-zone. The Z-line is located in the middle of the I-band.

Fig. 2
Fig. 2

Nonlinear multimodal microscope setup. The OE and OC represent excitation and collection objectives, respectively; D1 and D2 are dichroic mirrors, M1 is a mirror, and SM represents two galvanometric scanning mirrors. Additionally, optical filters F1, F2, and F3 are placed in front of the PMT detectors to isolate wavelengths of interest. (See text for a more detailed description.)

Fig. 3
Fig. 3

(Multimedia online; ao.osa.org) SHG microscopy images of Drosophila melanogaster larval muscle [(a) and (c)] and adult indirect flight muscle [(b) and (d)]. Two-dimensional images of the (a) larval and (b) adult muscle are presented. The stacks of images were rendered for the (c) larval and (d) adult myocytes. A conventional fluorescence image of (larval) muscle with actin labeled by rhodamine-conjugated phalloidin is shown in (e) for comparison. Images (a)–(d) were taken with 1042 nm excitation using a 1.3 NA oil immersion objective. Scale bar represents 5 μm in (a)–(d) and 100 μm in (e).

Fig. 4
Fig. 4

Comparison of SHG and MPF signals from Drosophila melanogaster larvae muscle. Typical images from Drosophila larvae muscle recorded with (a) SHG and (b) MPF microscopy at 800 nm excitation. A profile of each signal across the row of sarcomeres, the line shown in (a) and (b), is presented in (c). The solid curve with circles represents the SHG profile; the dotted curve with open triangles shows the MPF profile. Images were taken with 800 nm excitation using a 0.75 NA air objective.

Fig. 5
Fig. 5

Analysis of SHG from IFM myocytes of different Drosophila melanogaster mutants. (a)–(d) correspond to SHG images: (a) wild-type, (b) Y97, (c) M h c 10 ; Y97, and (d) M h c 10 . For comparison, confocal images of similar preparations of Drosophila myocytes labeled with rhodamine-conjugated phalloidin are also presented: (e) wild-type, (f) Y97, (g) M h c 10 ; Y97, and (h) M h c 10 mutants. The contrast has been adjusted in (a)–(h) for structural visualization. For intensity comparison, a bar graph (i) shows the average SHG intensities for the wild-type and the different mutants. The intensity of SHG was calculated by averaging the SHG signals from various scans. Note that the background was thresholded, and zero values were ignored in calculating the average signals. Images were taken with 1042 nm excitation using a 0.75 NA air objective. The bar in the images corresponds to 5 μm .

Fig. 6
Fig. 6

(Multimedia online; ao.osa.org) Effect of sustained contraction on the neighboring sarcomeres. The SHG images at various times during sustained contraction of Drosophila melanogaster larvae muscle are shown. The elapsed times for each image are indicated in the upper right corner. The region of sustained contraction is circled by the oval, and the arrow points to the middle of the highly deformed region. Images were taken with 800 nm excitation using a 0.75 NA air objective.

Fig. 7
Fig. 7

(Multimedia online; ao.osa.org) The evolution of SHG intensity in Drosophila melanogaster larval myocytes after addition of KCl. (a) SHG intensity changes as the sample undergoes a loss of viability. A 2D image of the myocyte is presented at (b) the initial state and (c) after 90 min. The 3D rendered structures of the muscle at (d) the initial state, and (e) after 90 min show significant structural changes. Images were taken with 1042 nm excitation using a 0.75 NA air objective. Scale bar represents 15 μm .

Fig. 8
Fig. 8

Influence of periodic contractions in the Drosophila melanogaster myocyte on the SHG efficiency. Changes in sarcomere length during spontaneous periodic contractions are shown with triangles, while SHG intensity changes are shown with squares. Data were collected with 800 nm excitation using a 0.75 NA air objective.

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