Abstract

We studied the polarization anisotropy of second harmonic generation (SHG) in polyhedral inclusion bodies (PIBs) of nuclear polyhedrosis viruses (NPV). Due to a body-centered-cubic arrangement of polyhedrin trimers, a characteristic SHG polarization property with a mixture of I23 and I3 symmetry was measured from PIBs. With this characteristic SHG anisotropy, it provides an intrinsic nonlinear signal for virus infection studies in living cells. With multimodal harmonic generation microscopy, we also demonstrated 3D imaging on PIBs of NPV in living cells. The distribution and the number of PIBs in intact infected cells can be revealed without the help of fluorescent labeling.

© 2008 Optical Society of America

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References

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  1. J. H. Chang, J. Y. Choi, B. R. Jin, J. Y. Roh, J. A. Olszewski, S. J. Seo, D. R. O'Reilly, and Y. H. Je, "An improved baculovirus insecticide producing occlusion bodies that contain Bacillus thuringiensis insect toxin," J. Invertebr. Pathol. 84, 30-37 (2003).
    [CrossRef] [PubMed]
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    [CrossRef]
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  4. C.-K. Sun, S.-W. Chu, S.-Y. Chen, T.-H. Tsai, T.-M. Liu, C.-Y. Lin, and H.-J. Tsai, "Higher harmonic generation microscopy for developmental biology," J. Struct. Biol. 147, 19- 30 (2004).
    [CrossRef] [PubMed]
  5. Asian Vegetable Research and Development Center (AVRDC). AVRDC Annual Report 2004, W. Easdown and T. Kalb, eds., (AVRDC, Shanhua, Tainan, Taiwan, 2007).
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    [CrossRef] [PubMed]
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    [CrossRef] [PubMed]
  11. F. Coulibaly, E. Chiu, K. Ikeda, S. Gutmann, P. W. Haebel, C. Schulze-Briese, H. Mori, and P. Metcalf, "The molecular organization of cypovirus polyhedra," Nature 446, 97-101 (2007).
    [CrossRef] [PubMed]
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    [CrossRef]

2007 (3)

S. C. Yeh, S. T. Lee, C Y. Wu, and C. H. Wang. "A cell line (NTU-MV) established from Maruca vitrata (Lepidoptera: pyralidae): characterization, viral susceptibility, and polyhdra production," J. Invertebr. Pathol. 96, 138-146 (2007).
[CrossRef] [PubMed]

F. Coulibaly, E. Chiu, K. Ikeda, S. Gutmann, P. W. Haebel, C. Schulze-Briese, H. Mori, and P. Metcalf, "The molecular organization of cypovirus polyhedra," Nature 446, 97-101 (2007).
[CrossRef] [PubMed]

S.-W. Chu, S.-P. Tai, C.-K. Sun, and C.-H. Lin, "Selective imaging in second-harmonic-generation microscopy by polarization manipulation," Appl. Phys. Lett. 91, 103903 (2007).
[CrossRef]

2006 (1)

S.-Y. Chen, C.-S. Hsieh, S.-W. Chu, C.-Y. Lin, C.-Y. Ko, Y.-C. Chen, H.-J. Tsai, C.-H. Hu, and C.-K. Sun, "Noninvasive harmonics optical microscopy for long-term observation of embryonic nervous system development in vivo," J. Biomed. Opt. 11, 054022 (2006).
[CrossRef] [PubMed]

2005 (3)

R. M. Williams, W. R. Zipfel, and W. W. Webb, "Interpreting second-harmonic generation images of collagen I fibrils," Biophys. J. 88, 1377-1386 (2005).
[CrossRef]

T.-C. Lee, S.-M. Yu, and Y. C. Chao. "Fusion of foreign protein genes to the occlusion body gene of the baculovirus to assist protein isolation and antibody production," J. Genet. Mol. Biol. 16, 138-150 (2005).

K. Anduleit, G. Sutton, J. M. Diprose, P. P. C. Mertens, J. M. Grimes, and D. I. Stuart, "Crystal lattice as biological phenotype for insect viruses," Protein Sci. 14, 2741-2743 (2005).
[CrossRef] [PubMed]

2004 (1)

C.-K. Sun, S.-W. Chu, S.-Y. Chen, T.-H. Tsai, T.-M. Liu, C.-Y. Lin, and H.-J. Tsai, "Higher harmonic generation microscopy for developmental biology," J. Struct. Biol. 147, 19- 30 (2004).
[CrossRef] [PubMed]

2003 (1)

J. H. Chang, J. Y. Choi, B. R. Jin, J. Y. Roh, J. A. Olszewski, S. J. Seo, D. R. O'Reilly, and Y. H. Je, "An improved baculovirus insecticide producing occlusion bodies that contain Bacillus thuringiensis insect toxin," J. Invertebr. Pathol. 84, 30-37 (2003).
[CrossRef] [PubMed]

2002 (1)

1996 (1)

P. J. Harshman, T. K. Gustafson, and P. Kelley, "Baculovirus-mediated gene transfer into mammalian cells," Proc. Natl. Acad. Sci. U.S.A. 93, 2348-2352 (1996).
[CrossRef]

Appl. Phys. Lett. (1)

S.-W. Chu, S.-P. Tai, C.-K. Sun, and C.-H. Lin, "Selective imaging in second-harmonic-generation microscopy by polarization manipulation," Appl. Phys. Lett. 91, 103903 (2007).
[CrossRef]

Biophys. J. (1)

R. M. Williams, W. R. Zipfel, and W. W. Webb, "Interpreting second-harmonic generation images of collagen I fibrils," Biophys. J. 88, 1377-1386 (2005).
[CrossRef]

J. Biomed. Opt. (1)

S.-Y. Chen, C.-S. Hsieh, S.-W. Chu, C.-Y. Lin, C.-Y. Ko, Y.-C. Chen, H.-J. Tsai, C.-H. Hu, and C.-K. Sun, "Noninvasive harmonics optical microscopy for long-term observation of embryonic nervous system development in vivo," J. Biomed. Opt. 11, 054022 (2006).
[CrossRef] [PubMed]

J. Genet. Mol. Biol. (1)

T.-C. Lee, S.-M. Yu, and Y. C. Chao. "Fusion of foreign protein genes to the occlusion body gene of the baculovirus to assist protein isolation and antibody production," J. Genet. Mol. Biol. 16, 138-150 (2005).

J. Invertebr. Pathol. (2)

J. H. Chang, J. Y. Choi, B. R. Jin, J. Y. Roh, J. A. Olszewski, S. J. Seo, D. R. O'Reilly, and Y. H. Je, "An improved baculovirus insecticide producing occlusion bodies that contain Bacillus thuringiensis insect toxin," J. Invertebr. Pathol. 84, 30-37 (2003).
[CrossRef] [PubMed]

S. C. Yeh, S. T. Lee, C Y. Wu, and C. H. Wang. "A cell line (NTU-MV) established from Maruca vitrata (Lepidoptera: pyralidae): characterization, viral susceptibility, and polyhdra production," J. Invertebr. Pathol. 96, 138-146 (2007).
[CrossRef] [PubMed]

J. Opt. Soc. Am. B (1)

J. Struct. Biol. (1)

C.-K. Sun, S.-W. Chu, S.-Y. Chen, T.-H. Tsai, T.-M. Liu, C.-Y. Lin, and H.-J. Tsai, "Higher harmonic generation microscopy for developmental biology," J. Struct. Biol. 147, 19- 30 (2004).
[CrossRef] [PubMed]

Nature (1)

F. Coulibaly, E. Chiu, K. Ikeda, S. Gutmann, P. W. Haebel, C. Schulze-Briese, H. Mori, and P. Metcalf, "The molecular organization of cypovirus polyhedra," Nature 446, 97-101 (2007).
[CrossRef] [PubMed]

Proc. Natl. Acad. Sci. U.S.A. (1)

P. J. Harshman, T. K. Gustafson, and P. Kelley, "Baculovirus-mediated gene transfer into mammalian cells," Proc. Natl. Acad. Sci. U.S.A. 93, 2348-2352 (1996).
[CrossRef]

Protein Sci. (1)

K. Anduleit, G. Sutton, J. M. Diprose, P. P. C. Mertens, J. M. Grimes, and D. I. Stuart, "Crystal lattice as biological phenotype for insect viruses," Protein Sci. 14, 2741-2743 (2005).
[CrossRef] [PubMed]

Other (2)

Asian Vegetable Research and Development Center (AVRDC). AVRDC Annual Report 2004, W. Easdown and T. Kalb, eds., (AVRDC, Shanhua, Tainan, Taiwan, 2007).

A. Yariv, Optical Electronics in Modern Communications, 5th ed. (Oxford University Press, 1997), pp. 330.

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Figures (5)

Fig. 1.
Fig. 1.

Combined image of second harmonic generation (SHG) (green color) and third harmonic generation (THG) (yellow color) from uninfected cells in the NTU-MV cell line. No SHG can be found in uninfected cells.

Fig. 2.
Fig. 2.

(a). Combined (b) SHG (green color) and (c) THG (yellow color) image of an infected cell.

Fig. 3.
Fig. 3.

Combined SHG (green color) and THG (yellow color) image of an infected cell.

Fig. 4.
Fig. 4.

(a). The SHG intensity along the X-axis versus the rotated angle of laser polarization. (b) The SHG intensity along the Y-axis versus the rotated angle of laser polarization. Black curves represent the fitting results from the symmetry analysis.

Fig. 5.
Fig. 5.

Coordinate assignment of the simplified trimer with its H4-helices (a) pointing out and (b) into the paper. (c) The arrangement of the trimers in the unit cubic cell of the PIB crystal. Red arrows represent the direction of x′-axes and blue the direction of y′-axes. The spatial arrangements of trimers in different planes are illustrated: (d) ABGH plane and (e) CDEF plane. Point O is the center of the cubic and green arrows represent the direction of z′-axis.

Equations (7)

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

d = tri = ( d 11 d 11 0 d 14 d 15 d 22 d 22 d 22 0 d 15 d 14 d 11 d 31 d 31 d 33 0 0 0 ) .
P loc ( 2 ω ) = ( P x ( 2 ω ) P y ( 2 ω ) P 2 ( 2 ω ) ) = d = tri ( E x 2 E y 2 E z 2 E y E z E x E z E x E y ) ,
( E x E y E z ) = ( 6 6 6 6 6 3 2 2 2 2 0 3 3 3 3 3 3 ) ( E x 1 E x 2 E x 3 ) = T ( E x 1 E x 2 E x 3 ) ,
P x 1 ( 2 ω ) = 16 9 ( 6 d 11 3 2 d 22 ) E x 1 E x 3 + 16 3 9 ( 2 d 15 d 31 + d 33 ) E x 2 E x 3 ,
P x 2 ( 2 ω ) = 16 9 ( 6 d 11 + 3 2 d 22 ) E x 2 E x 3 + 16 3 9 ( 2 d 15 d 31 + d 33 ) E x 1 E x 3 ,
P x 3 ( 2 ω ) = 8 9 ( 6 d 11 3 2 d 22 ) E x 1 2 + 8 9 ( 6 d 11 + 3 2 d 22 ) E x 2 2 16 6 9 d 11 E x 3 2 + 16 3 9 ( 2 d 15 d 31 + d 33 ) E x 1 E x 2 .
D = PIB = ( 0 0 0 D 14 D 15 0 0 0 0 D 24 D 14 0 D 15 2 D 24 2 D 33 0 0 D 14 ) .

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