04/22/04

Second harmonic generation microscopy

"A scanning optical microscope in which an image is produced from the generation of optical second harmonics within the specimen has been constructed. Pictures have been obtained from various crystals which show high contrast levels and detail not visible with the conventional microscope."

JN Gannaway, CJR Sheppard, Optical and Quantum Electronics, 10, 435-439 (1978)

Two-photon fluorescence microscopy

"The S[canning] O[ptical] M[icroscope] is well suited to spectroscopic applications such as generation of images from fluorescent  or luminescent radiation or from Raman scattering. By using a medium power laser it is possible to excite nonlinear optical effects ... Other possibilities include imaging using coherent Raman scattering, two-photon fluorescence and others."

CJR Sheppard, Proc. SPIE, 368, 88-95 (1983)

2 Multiphoton Microscopy

F. 5.     Kompfner, R., Sheppard, C.J.R., Walsh, D., Choudhury, A., Gannaway, J.N., Hale, P.G. (1977) Applications of quantum electronics, Part 1:  The scanning optical microscope, OUEL Report No. 1183/77.

E.6. Sheppard CJR, Kompfner R, Gannaway J, Walsh D (1977) The scanning harmonic optical microscope, IEEE/OSA Conf. Laser Engineering and Applications Washington, IEEE J. Quantum Elec., QE-13, 100D.

First report of scanning second harmonic generation microscopy.

Reproduced in SPIE Milestone Series MS-175, Selected reprints on Multiphoton excitation microscopy, BR Masters, ed. ISBN 0-8194-4748X, p. 33

D. 14. Sheppard CJR, Kompfner R (1978) Resonant scanning optical microscope, Appl. Opt. 17, 2879-2882 (and cover).

First proposal of two-photon fluorescence microscopy.

Reproduced in SPIE Milestone Series MS-175, Selected reprints on Multiphoton excitation microscopy, BR Masters, ed. ISBN 0-8194-4748X, pp.  39-42

“In the scanning optical microscope nonlinear interactions are expected to occur between object and a highly focused beam of light, which we hope will open new ways of studying matter in microscopic detail hitherto not available. Nonlinear interactions include the generation of sum frequencies, Raman scattering, two-photon fluorescence, and others. We feel that the method will be of particular interest in studying biological materials … Furthermore, frequency mixing should give information concerning the chemical structure of the object.

However, heating in the object sets a limit to the amount of power that can be used because some fraction of it will be absorbed and transformed into heat. We distinguish between two kinds of scanning: slow and fast. In the slow-scan case, where the probe is effectively stationary on the object, we estimate that the permissible optical beam power is about 35mW. In the fast-scan case, the whole specimen reaches an equilibrium temperature, and the power is limited to about 100mW. It is clear that heat dissipation is a serious problem for the harmonic scanning optical microscope, and ways of improving either the efficiency of conversion of fundamental to harmonic power, for example by beam pulsing, or the thermal conductance of the object supports of the object are of importance.”

D. 19. Gannaway J, Sheppard CJR (1978) Second harmonic imaging in the scanning optical microscope, Opt. and Quant. Elec. 10, 435-439.

First published second harmonic generation microscopy images.

Reproduced in SPIE Milestone Series MS-131, Selected reprints on Confocal microscopy, BR Masters, ed. ISBN 0-8194-2376-6, pp, 406-410

Reproduced in SPIE Milestone Series MS-175, Selected reprints on Multiphoton excitation microscopy, BR Masters, ed. ISBN 0-8194-4748X, pp. 34-38

D. 23. Wilson T, Sheppard CJR (1979) Imaging and super-resolution in the harmonic micrsocope, Optica Acta 26, 761-770.

Imaging in a harmonic microscope is partially-coherent.

D. 97. Sheppard CJR, Gu M (1990) Image formation in two-photon fluorescence microscopy, Optik 86, 104-106.

Reproduced in SPIE Milestone Series MS-175, Selected reprints on Multiphoton excitation microscopy, BR Masters, ed. ISBN 0-8194-4748X, pp. 127-136

D. 139. Gu M, Sheppard CJR (1993) Effects of a finite-sized pinhole on 3-D image formation in confocal two-photon fluorescence microscopy, J. Mod. Opt. 40, 2009-2024.

D. 162. Gu M, Sheppard CJR (1995) Optical transfer function analysis for two-photon 4Pi confocal fluorescence microscopy, Opt. Commun. 114, 45-49

D. 175. Gu M, Sheppard CJR (1995) Comparison of three-dimensional imaging properties between two-photon and single-photon fluorescence microscopy, J. Microsc. 177, 128-137

D. 191. Sheppard CJR (1996) Imaging in three-photon fluorescence microscopy, Bioimaging, 4 124-128

D. 209. Gauderon R, Lukins PB and Sheppard CJR (1998) Three-dimensional second-harmonic generation imaging with femtosecond laser pulses, Opt. Lett. 23, 1209-1211

C. 26. Cox G and Sheppard CJR (1999) Multiphoton fluorescence microscopy, in Fluorescent and Luminescent Probes, 2nd. ed., WT Mason, ed., Academic Press, London, pp.331-336. ISBN 0 12 447836 0

D. 226. Gauderon R, Lukins PB and Sheppard CJR (1999) Effect of a confocal pinhole in two-photon microscopy, Microsc. Res. and Tech. 47, 210-214

D. 230. Gauderon R, Sheppard CJR (1999) Effect of a finite-sized pinhole on noise performance in single-, two-, and three-photon confocal fluorescence microscopy, Appl. Opt. 38, 3562-3565

E.80. Gauderon R, Lukins PB, Sheppard CJR (2000) Simultaneous multichannel nonlinear imaging: combined two-photon excited fluorescence and second-harmonic generation microscopy, Optical sensisng, imaging and manipulation for biological and biomedical applications, Photonics Taiwan, Taipei 26-28 July, Proc. SPIE 4082 115-118

D. 234. Gauderon R, Sheppard CJR (2000) Two-dimensional weak-object transfer functions in the scanning harmonic microscope, J. Mod. Opt. 47, 1195-1202

D. 245. Gauderon R, Lukins PB, Sheppard CJR (2001) Optimisation of second-harmonic generation microscopy, Micron, 32, 691-700

D. 246. Gauderon R, Lukins PB, Sheppard CJR (2001) Simultaneous multi-channel two- photon microscopy, Micron 32, 685-689

C. 34. Sheppard CJR (2002) The generalized microscope, in Confocal and two-photon microscopy: Foundations, applications, and advances, ed. A Diaspro, Wiley-Liss, New York, ISBN 0-471-40920-0, pp1-18

C. 35. Diaspro A, Sheppard CJR (2002) Two-photon microscopy: Basic principles and architectures, in Confocal and two-photon microscopy: Foundations, applications, and advances, ed. A Diaspro, Wiley-Liss, New York, ISBN 0-471-40920-0, pp 39-74

C. 36. Török P, Sheppard CJR (2002) The role of pinhole size in high-aperture two- and three-photon microscopy, in Confocal and two-photon microscopy: Foundations, applications, and advances, ed. A Diaspro, Wiley-Liss, New York, ISBN 0-471-40920-0, pp127-152

 E. 88 Cox GC, Kable E, Sheppard CJR, Xu P (2002) Resolution in second harmonic generation microscopy, 15th Int. Congress on Electron Microscopy, Durban, South Africa, 1-6 September, Proc. ICEM-15, 2, 331-332

E. 93 Cox G, Xu P, Sheppard CJR, Ramshaw J (2003) Characterization of the Second Harmonic Signal from Collagen, San Jose. Proc. SPIE 4963, 32-40