Observation of stimulated emission by direct three-photon excitation

He, Guang S.; Markowicz, Przemyslaw P.; Lin, Tzu Chau; Prasad, Paras N.

doi:10.1038/415767a

Cited by 374 publications

(237 citation statements)

References 28 publications

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“…In this paper, we propose a method to perform 2PE-STED imaging using a single wavelength (SW) and, consequently, the very same laser source for 2PE and depletion. Such a SW approach is in agreement with early reported spectral evidences for different dyes (11,12).…”

supporting

confidence: 79%

Single-wavelength two-photon excitation–stimulated emission depletion (SW2PE-STED) superresolution imaging

Bianchini

Harke

Galiani

et al. 2012

Proc. Natl. Acad. Sci. U.S.A.

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We developed a new class of two-photon excitation-stimulated emission depletion (2PE-STED) optical microscope. In this work, we show the opportunity to perform superresolved fluorescence imaging, exciting and stimulating the emission of a fluorophore by means of a single wavelength. We show that a widely used redemitting fluorophore, ATTO647N, can be two-photon excited at a wavelength allowing both 2PE and STED using the very same laser source. This fact opens the possibility to perform 2PE microscopy at four to five times STED-improved resolution, while exploiting the intrinsic advantages of nonlinear excitation.T wo-photon excitation (2PE) fluorescence microscopy (1) is a widely used technique in medicine and biology (2, 3). Its characteristics make it particularly suitable for deep tissue and in vivo imaging applications (4). The use of infrared wavelengths guarantees a high penetration depth in scattering samples and a reduced photodamage in living specimens. Moreover, the quadratic dependence of the excitation allows intrinsical rejection of the out-of-focus fluorescence background and reduces the FWHM of the focal spot by approximately ffiffiffi 2 p compared to the diffraction limited spot of excitation light. However, 2PE cannot be considered a genuine superresolution technique: The "doubling" in wavelength causes that the size of the diffraction spot of excitation light is twice than the one achievable by single-photon excitation (1PE). In 1994, Hell and Wichmann introduced the stimulated emission depletion (STED) concept as a way to achieve optical superresolution by breaking the diffraction barrier (5). In general, STED imaging is based on the fluorescence switching due to stimulated emission depletion introduced by a second red shifted beam. The fluorescent sample is probed by a focused excitation beam overlaid with a STED beam commonly shaped to a doughnut-like intensity distribution featuring zero intensity in the center. Because of the saturation of the stimulated emission depletion, the fluorescence is switched in the whole focal area except for the small region around the zero of the STED beam. The resulting resolution is theoretically unlimited being governed by the efficiency of the depletion process (6, 7). Recently, STED has been proposed coupled with 2PE microscopy combining the advantages of 2PE with its superresolution ability (8, 9). In general, continuous wave (CW)-STED (10) is preferred for 2PE imaging due to a simplification of the experimental setup at the expenses of an increased cost. Unfortunately, the use of two distinct wavelengths for excitation and depletion requires mostly a special optical filter design making the setup invariable in terms of the choice of the marker dye, and potential light beam distortions have to be treated separately. Working with only just one wavelength for both excitation and STED, one directly simplifies the image formation scheme. In this paper, we propose a method to perform 2PE-STED imaging using a single wavelength (SW) and, consequently, the very...

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supporting

confidence: 79%

Single-wavelength two-photon excitation–stimulated emission depletion (SW2PE-STED) superresolution imaging

Bianchini

Harke

Galiani

et al. 2012

Proc. Natl. Acad. Sci. U.S.A.

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“…With the development of laser technology, three-photon excited upconversion, especially simultaneous one, has gained much interest due to its longer excitation wavelength and stronger spatial confinement based on the cubic dependence of the three-photon process on the input light intensity. Currently, almost all reports on up-converted luminescence by three-photon simultaneous absorption have been concentrated on organic materials [12,13]. Only a little attention has been paid to the similar phenomenon in inorganic solid-state materials [14,15], especially in crystals.…”

Section: Introductionmentioning

confidence: 99%

Simultaneous three-photon absorption induced ultraviolet upconversion in Pr3+:Y2SiO5 crystal by femtosecond laser irradiation

Wang¹,

Qiu²,

Song³

et al. 2008

Optics Communications

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“…where M (6) is a 15 × 15 matrix and where the 15 elements of the vectors f (6) and g (6) are Kronecker delta strings. The expressions of f (6) , M (6) , and g (6) from Ref.…”

Section: Rotational Averaging Of Tensorsmentioning

confidence: 99%

“…5 These effects are of particular interest for frequency upconversion of infrared lasers to the visible and UV parts of the spectrum. 6 Applications of multiphoton absorption require not only strong lasers but also molecular materials with large nonlinear optical absorption cross sections. 7,8 To support the design of these materials, the development of theoretical methods for the treatment of nonlinear absorption is of large interest.…”

Section: Introductionmentioning

confidence: 99%

Rotational averaging of multiphoton absorption cross sections

Friese

Beerepoot

Ruud

2014

The Journal of Chemical Physics

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Articles you may be interested inRotational averaging of tensors is a crucial step in the calculation of molecular properties in isotropic media. We present a scheme for the rotational averaging of multiphoton absorption cross sections. We extend existing literature on rotational averaging to even-rank tensors of arbitrary order and derive equations that require only the number of photons as input. In particular, we derive the first explicit expressions for the rotational average of five-, six-, and seven-photon absorption cross sections. This work is one of the required steps in making the calculation of these higher-order absorption properties possible. The results can be applied to any even-rank tensor provided linearly polarized light is used.

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Observation of stimulated emission by direct three-photon excitation

Cited by 374 publications

References 28 publications

Single-wavelength two-photon excitation–stimulated emission depletion (SW2PE-STED) superresolution imaging

Single-wavelength two-photon excitation–stimulated emission depletion (SW2PE-STED) superresolution imaging

Simultaneous three-photon absorption induced ultraviolet upconversion in Pr3+:Y2SiO5 crystal by femtosecond laser irradiation

Rotational averaging of multiphoton absorption cross sections

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