Abstract

We extend second-harmonic generation (SHG) microscopy to the measurement of sarcomere length in unstained living cardiac myocytes with 20-nm accuracy. We quantify individual sarcomere shortening in the presence of saxitoxin and find that it is in agreement with mechanical measurements of atrial tissue contracture. This functional application of SHG microscopy is generally applicable to quantify the physiological effects of drugs on contractile tissue. Our data also suggest that packed myosin heads in sarcomere thick filaments are responsible for the large second-harmonic endogenous signal in muscle tissue.

© 2004 Optical Society of America

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    [CrossRef] [PubMed]
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2003 (3)

W. R. Zipfel, R. M. Williams, R. Christie, A. Yu Nikitin, B. T. Hyman, and W. W. Webb, Proc. Natl. Acad. Sci. USA 100, 7075 (2003).
[CrossRef]

M. Both, M. Vogel, R. H. A. Fink, and D. Uttenweiler, Proc. SPIE 5139, 112 (2003).
[CrossRef]

S.-W. Chu, T.-M. Liu, C.-K. Sun, C.-Y. Lin, and H. J. Tsai, Opt. Express 11, 933 (2003), http://www.opticsexpress.org .
[CrossRef] [PubMed]

2002 (4)

P. J. Campagnola, A. C. Millard, M. Terasaki, P. E. Hoppe, C. J. Malone, and W. A. Mohler, Biophys. J. 82, 493 (2002).
[CrossRef]

R. Littlefield and V. M. Fowler, Biophys. J. 82, 2548 (2002).
[CrossRef] [PubMed]

M.-P. Sauviat, A. Colas, and N. Pages, BMC Pharmacol. 2, 15 (2002).
[CrossRef]

R. E. Thompson, D. R. Larson, and W. W. Webb, Biophys. J. 82, 2775 (2002).
[CrossRef] [PubMed]

2000 (2)

1999 (1)

P. J. Campagnola, M.-de Wei, A. Lewis, and L. M. Loew, Biophys. J. 77, 3341 (1999).
[CrossRef] [PubMed]

1998 (1)

P. W. Brandt, F. Colomo, N. Piroddi, C. Poggesi, and C. Tesi, Biophys. J. 74, 1994 (1998).
[CrossRef] [PubMed]

1997 (2)

Alfano, R. R.

Blanchard-Desce, M.

Both, M.

M. Both, M. Vogel, R. H. A. Fink, and D. Uttenweiler, Proc. SPIE 5139, 112 (2003).
[CrossRef]

Brandt, P. W.

P. W. Brandt, F. Colomo, N. Piroddi, C. Poggesi, and C. Tesi, Biophys. J. 74, 1994 (1998).
[CrossRef] [PubMed]

Campagnola, P. J.

P. J. Campagnola, A. C. Millard, M. Terasaki, P. E. Hoppe, C. J. Malone, and W. A. Mohler, Biophys. J. 82, 493 (2002).
[CrossRef]

P. J. Campagnola, M.-de Wei, A. Lewis, and L. M. Loew, Biophys. J. 77, 3341 (1999).
[CrossRef] [PubMed]

Christie, R.

W. R. Zipfel, R. M. Williams, R. Christie, A. Yu Nikitin, B. T. Hyman, and W. W. Webb, Proc. Natl. Acad. Sci. USA 100, 7075 (2003).
[CrossRef]

Chu, S.-W.

Colas, A.

M.-P. Sauviat, A. Colas, and N. Pages, BMC Pharmacol. 2, 15 (2002).
[CrossRef]

Colomo, F.

P. W. Brandt, F. Colomo, N. Piroddi, C. Poggesi, and C. Tesi, Biophys. J. 74, 1994 (1998).
[CrossRef] [PubMed]

Fink, R. H. A.

M. Both, M. Vogel, R. H. A. Fink, and D. Uttenweiler, Proc. SPIE 5139, 112 (2003).
[CrossRef]

Fowler, V. M.

R. Littlefield and V. M. Fowler, Biophys. J. 82, 2548 (2002).
[CrossRef] [PubMed]

Guo, Y.

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, Biophys. J. 82, 493 (2002).
[CrossRef]

Hyman, B. T.

W. R. Zipfel, R. M. Williams, R. Christie, A. Yu Nikitin, B. T. Hyman, and W. W. Webb, Proc. Natl. Acad. Sci. USA 100, 7075 (2003).
[CrossRef]

Larson, D. R.

R. E. Thompson, D. R. Larson, and W. W. Webb, Biophys. J. 82, 2775 (2002).
[CrossRef] [PubMed]

Lewis, A.

P. J. Campagnola, M.-de Wei, A. Lewis, and L. M. Loew, Biophys. J. 77, 3341 (1999).
[CrossRef] [PubMed]

Lin, C.-Y.

Littlefield, R.

R. Littlefield and V. M. Fowler, Biophys. J. 82, 2548 (2002).
[CrossRef] [PubMed]

Liu, F.

Liu, T.-M.

Loew, L. M.

P. J. Campagnola, M.-de Wei, A. Lewis, and L. M. Loew, Biophys. J. 77, 3341 (1999).
[CrossRef] [PubMed]

Malone, C. J.

P. J. Campagnola, A. C. Millard, M. Terasaki, P. E. Hoppe, C. J. Malone, and W. A. Mohler, Biophys. J. 82, 493 (2002).
[CrossRef]

Mertz, J.

Millard, A. C.

P. J. Campagnola, A. C. Millard, M. Terasaki, P. E. Hoppe, C. J. Malone, and W. A. Mohler, Biophys. J. 82, 493 (2002).
[CrossRef]

Mohler, W. A.

P. J. Campagnola, A. C. Millard, M. Terasaki, P. E. Hoppe, C. J. Malone, and W. A. Mohler, Biophys. J. 82, 493 (2002).
[CrossRef]

Moreaux, L.

Narahashi, T.

T. Narahashi, J. Pharmacol. Exp. Ther. 294, 1 (2000).
[PubMed]

Nikitin, A. Yu

W. R. Zipfel, R. M. Williams, R. Christie, A. Yu Nikitin, B. T. Hyman, and W. W. Webb, Proc. Natl. Acad. Sci. USA 100, 7075 (2003).
[CrossRef]

Pages, N.

M.-P. Sauviat, A. Colas, and N. Pages, BMC Pharmacol. 2, 15 (2002).
[CrossRef]

Piroddi, N.

P. W. Brandt, F. Colomo, N. Piroddi, C. Poggesi, and C. Tesi, Biophys. J. 74, 1994 (1998).
[CrossRef] [PubMed]

Poggesi, C.

P. W. Brandt, F. Colomo, N. Piroddi, C. Poggesi, and C. Tesi, Biophys. J. 74, 1994 (1998).
[CrossRef] [PubMed]

Sacks, P.

Sandre, O.

Sauviat, M.-P.

M.-P. Sauviat, A. Colas, and N. Pages, BMC Pharmacol. 2, 15 (2002).
[CrossRef]

Savage, H.

Schantz, S.

Squire, J. M.

J. M. Squire, Curr. Opin. Struct. Biol. 7, 247 (1997).
[CrossRef] [PubMed]

Sun, C.-K.

Terasaki, M.

P. J. Campagnola, A. C. Millard, M. Terasaki, P. E. Hoppe, C. J. Malone, and W. A. Mohler, Biophys. J. 82, 493 (2002).
[CrossRef]

Tesi, C.

P. W. Brandt, F. Colomo, N. Piroddi, C. Poggesi, and C. Tesi, Biophys. J. 74, 1994 (1998).
[CrossRef] [PubMed]

Thompson, R. E.

R. E. Thompson, D. R. Larson, and W. W. Webb, Biophys. J. 82, 2775 (2002).
[CrossRef] [PubMed]

Tsai, H. J.

Uttenweiler, D.

M. Both, M. Vogel, R. H. A. Fink, and D. Uttenweiler, Proc. SPIE 5139, 112 (2003).
[CrossRef]

Vogel, M.

M. Both, M. Vogel, R. H. A. Fink, and D. Uttenweiler, Proc. SPIE 5139, 112 (2003).
[CrossRef]

Webb, W. W.

W. R. Zipfel, R. M. Williams, R. Christie, A. Yu Nikitin, B. T. Hyman, and W. W. Webb, Proc. Natl. Acad. Sci. USA 100, 7075 (2003).
[CrossRef]

R. E. Thompson, D. R. Larson, and W. W. Webb, Biophys. J. 82, 2775 (2002).
[CrossRef] [PubMed]

Wei, M.-de

P. J. Campagnola, M.-de Wei, A. Lewis, and L. M. Loew, Biophys. J. 77, 3341 (1999).
[CrossRef] [PubMed]

Williams, R. M.

W. R. Zipfel, R. M. Williams, R. Christie, A. Yu Nikitin, B. T. Hyman, and W. W. Webb, Proc. Natl. Acad. Sci. USA 100, 7075 (2003).
[CrossRef]

Zhadin, N.

Zipfel, W. R.

W. R. Zipfel, R. M. Williams, R. Christie, A. Yu Nikitin, B. T. Hyman, and W. W. Webb, Proc. Natl. Acad. Sci. USA 100, 7075 (2003).
[CrossRef]

Biophys. J. (5)

P. J. Campagnola, M.-de Wei, A. Lewis, and L. M. Loew, Biophys. J. 77, 3341 (1999).
[CrossRef] [PubMed]

R. Littlefield and V. M. Fowler, Biophys. J. 82, 2548 (2002).
[CrossRef] [PubMed]

R. E. Thompson, D. R. Larson, and W. W. Webb, Biophys. J. 82, 2775 (2002).
[CrossRef] [PubMed]

P. W. Brandt, F. Colomo, N. Piroddi, C. Poggesi, and C. Tesi, Biophys. J. 74, 1994 (1998).
[CrossRef] [PubMed]

P. J. Campagnola, A. C. Millard, M. Terasaki, P. E. Hoppe, C. J. Malone, and W. A. Mohler, Biophys. J. 82, 493 (2002).
[CrossRef]

BMC Pharmacol. (1)

M.-P. Sauviat, A. Colas, and N. Pages, BMC Pharmacol. 2, 15 (2002).
[CrossRef]

Curr. Opin. Struct. Biol. (1)

J. M. Squire, Curr. Opin. Struct. Biol. 7, 247 (1997).
[CrossRef] [PubMed]

J. Pharmacol. Exp. Ther. (1)

T. Narahashi, J. Pharmacol. Exp. Ther. 294, 1 (2000).
[PubMed]

Opt. Express (1)

Opt. Lett. (2)

Proc. Natl. Acad. Sci. USA (1)

W. R. Zipfel, R. M. Williams, R. Christie, A. Yu Nikitin, B. T. Hyman, and W. W. Webb, Proc. Natl. Acad. Sci. USA 100, 7075 (2003).
[CrossRef]

Proc. SPIE (1)

M. Both, M. Vogel, R. H. A. Fink, and D. Uttenweiler, Proc. SPIE 5139, 112 (2003).
[CrossRef]

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Figures (3)

Fig. 1
Fig. 1

Schematic representation of a sarcomere (see text).

Fig. 2
Fig. 2

SHG images from unstained atrial myocytes freshly isolated from an adult frog heart and kept in a Ringer solution. The sarcomeres are clearly delimited by thick black lines, and the SHG signal originates from the myosin filaments. A reduced SHG signal is obtained from the bare central region of the myosin filaments, which appears as a thin black line. A, myocytes in Ringer solution; B, myocytes in Ringer solution containing STX (8 nM). Excitation, 860 nm, 20 mW; 0.9-N.A. water objective. Image acquisition time, 2.3 s 390×295 pixels/50 kHz.

Fig. 3
Fig. 3

SHG signal profile along a single atrial myocyte isolated from an adult frog heart. A, SHG image of a few sarcomeres selected in Figs. 2A and 2B; B, SHG signal (squares) extracted along a straight line in A. The solid curves correspond to fitting with multiple Gaussians. Individual Gaussians are also displayed to visualize the center of the peak (see solid vertical lines). The double arrows indicate the lengths reported in Table 1.

Tables (1)

Tables Icon

Table 1 Sarcomere Characteristic Dimensions in the Absence and in the Presence of STX

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