Principles of Three-Dimensional Imaging in Confocal Microscopes

Gu, Min

doi:10.1142/9789814261104

Cited by 107 publications

(91 citation statements)

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“…In the case of 1p excitation, a pinhole 300 m ͑optical unit ϳ3) in diameter was placed in front of the detector to produce an optical sectioning effect with strength similar to that under 2p excitation without using a pinhole. 1,13 This arrangement implies that the ability of rejecting scattered photons caused by the optical sectioning effect is the same in the two cases. 14 Figure 2 shows the dependence of the measured fluorescence signal level on the sample thickness d under 1p and 2p excitation.…”

Section: ͓S0003-6951͑00͒04536-8͔mentioning

confidence: 95%

Comparison of penetration depth between two-photon excitation and single-photon excitation in imaging through turbid tissue media

Gan

Kisteman

et al. 2000

Applied Physics Letters

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We show, both theoretically and experimentally, that for a turbid tissue medium where Mie scattering is dominant, multiple scattering not only reduces the illumination power in the forward direction but also exhibits an anisotropic distribution of scattered photons. Thus, a signal level under two-photon excitation drops much faster than that under single-photon excitation although image resolution is much higher in the former case. As a result, the penetration depth under two-photon excitation is limited by the strength of two-photon fluorescence and is not necessarily larger than that under single-photon excitation.

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Section: ͓S0003-6951͑00͒04536-8͔mentioning

confidence: 95%

Comparison of penetration depth between two-photon excitation and single-photon excitation in imaging through turbid tissue media

Gan

Kisteman

et al. 2000

Applied Physics Letters

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“…The 3-D PSF has an ellipsoidal shape elongated along the optical axis [23,24]. Thus, the obtained confocal image is the convolution of actual intensity distributions using the PSF as a kernel.…”

Section: Dvc Using Confocal Microscope Imagesmentioning

confidence: 99%

Three-dimensional Full-field Measurements of Large Deformations in Soft Materials Using Confocal Microscopy and Digital Volume Correlation

et al. 2007

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A three-dimensional (3-D) full-field measurement technique was developed for measuring large deformations in optically transparent soft materials. The technique utilizes a digital volume correlation (DVC) algorithm to track motions of subvolumes within 3-D images obtained using fluorescence confocal microscopy. In order to extend the strain measurement capability to the large deformation regime (>5%), a stretch-correlation algorithm was developed and implemented into the Fast Fourier Transform (FFT)-based DVC algorithm. The stretch-correlation algorithm uses a logarithmic coordinate transformation to convert the stretchcorrelation problem into a translational correlation problem under the assumption of small rotation and shear. Estimates of the measurement precision are provided by stationary and translation tests. The proposed measurement technique was used to measure large deformations in a transparent agarose gel sample embedded with fluorescent particles under uniaxial compression. The technique was also employed to measure non-uniform deformation fields near a hard spherical inclusion under far-field uniaxial compression. Introduction of the stretch-correlation algorithm greatly improved the strain measurement accuracy by providing better precision especially under large deformation. Also, the deconvolution of confocal images improved the accuracy of the measurement in the direction of the optical axis. These results shows that the proposed technique is well-suited for investigating cell-matrix mechanical interactions as well as for obtaining local constitutive properties of soft biological materials including tissues in 3-D.

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“…Nevertheless, utilizing the analogous reasoning applied to the field of incoherent fluorescent CSOM [25][26][27], to obtain estimates for incoherent SCEM resolution limits we assume circumstances in which the nonlocal potential in Eq. (22) is proportional to dðr ?…”

Section: Point Spread Functions In Stem and Scemmentioning

confidence: 99%

“…(26) and making the approximation that the free space STEM probe evolution given by Eq. (10) sufficiently describes the probe within the specimen we find that the Fourier transform of the recorded 3D STEM image is (see Appendix B)…”

Section: Article In Pressmentioning

confidence: 99%

“…The local STEM expression of Eq. (26) shows that, for a given probe position and defocus, the signal is the overlap integral between the probe intensity and the effective scattering potential. It is a direct consequence of the conservation of electrons that the integrated intensity is the same in all planes perpendicular to the beam axis.…”

Section: Article In Pressmentioning

confidence: 99%

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Three-dimensional imaging in double aberration-corrected scanning confocal electron microscopy, Part II: Inelastic scattering

et al. 2008

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Principles of Three-Dimensional Imaging in Confocal Microscopes

Cited by 107 publications

References 0 publications

Comparison of penetration depth between two-photon excitation and single-photon excitation in imaging through turbid tissue media

Comparison of penetration depth between two-photon excitation and single-photon excitation in imaging through turbid tissue media

Three-dimensional Full-field Measurements of Large Deformations in Soft Materials Using Confocal Microscopy and Digital Volume Correlation

Three-dimensional imaging in double aberration-corrected scanning confocal electron microscopy, Part II: Inelastic scattering

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