Line-scan focal modulation microscopy

Pant, Shilpa; Li, Caixia; Gong, Zhiyuan; Chen, Nanguang

doi:10.1117/1.jbo.22.5.050502

Cited by 15 publications

(17 citation statements)

References 29 publications

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“…This problem can be alleviated by introducing additional background rejection mechanisms beyond spatial filtering provided by confocal detection [41][42][43]. For example, focal modulation has been demonstrated to be very effective in suppressing scattering induced background in visible light confocal microscopy [44][45][46][47][48][49][50].…”

Section: Discussionmentioning

confidence: 99%

Three‐dimensional cellular imaging in thick biological tissue with confocal detection of one‐photon fluorescence in the near‐infrared II window

Wang

Chen

2019

Journal of Biophotonics

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Fluorescence imaging in the second near‐infrared optical window (NIR‐II, 900‐1700 nm) has become a technique of choice for noninvasive in vivo imaging in recent years. Greater penetration depths with high spatial resolution and low background can be achieved with this NIR‐II window, owing to low autofluorescence within this optical range and reduced scattering of long wavelength photons. Here, we present a novel design of confocal laser scanning microscope tailored for imaging in the NIR‐II window. We showcase the outstanding penetration depth of our confocal setup with a series of imaging experiments. HeLa cells labeled with PbS quantum dots with a peak emission wavelength of 1276 nm can be visualized through a 3.5‐mm‐thick layer of scattering medium, which is a 0.8% Lipofundin solution. A commercially available organic dye IR‐1061 (emission peak at 1132 nm), in its native form, is used for the first time, as a NIR‐II fluorescence label in cellular imaging. Our confocal setup is capable of capturing optically sectioned images of IR‐1061 labeled chondrocytes in fixed animal cartilage at a depth up to 800 μm, with a superb spatial resolution of around 2 μm.

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Section: Discussionmentioning

confidence: 99%

Three‐dimensional cellular imaging in thick biological tissue with confocal detection of one‐photon fluorescence in the near‐infrared II window

Wang

Chen

2019

Journal of Biophotonics

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“…This is not surprising given the numerous advantages it offers including cellular (i.e., micron-level) axial resolution, noninvasive nature, cost-effectiveness, and the ability to be incorporated into catheters [2] and endoscopes [3]. Unlike many other microscopic methods such as fluorescence microscopy [4], OCT does not require exogenous labeling for visualizing biological structures. Recently there has been a growing interest in applying OCT to pre-clinical studies that involve various small animal models.…”

Section: Introductionmentioning

confidence: 99%

Ultrahigh-speed line-scan SD-OCT for four-dimensional in vivo imaging of small animal models

Al-Qazwini¹,

Ko²,

Mehta³

et al. 2018

Biomed. Opt. Express

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We report an ultrahigh-speed and high-resolution line-scan spectral-domain optical coherence tomography (SD-OCT) system that integrates a number of mechanisms for improving image quality. The illumination uniformity is significantly improved by the use of a Powell lens; Phase stepping and differential reconstruction are combined to suppress autocorrelation artifacts; Nonlocal means (NLM) is employed to enhance the signal to noise ratio while minimizing motion artifacts. The system is capable of acquiring cross-sectional images at more than 3,500 B-scans per second with sensitivities between 70dB and 90dB. The high B-scan rate enables image post-processing with nonlocal means, an advanced noise reduction algorithm that affords enhanced morphological details and reduced motion artifacts. The achieved axial and lateral resolutions are 2.0 and 6.2 microns, respectively. We have used this system to acquire four-dimensional (three-dimensional space and one-dimensional time) imaging data from live chicken embryos at up to 40 volumes per second. Dynamic cardiac tissue deformation and blood flow could be clearly visualized at high temporal and spatial resolutions, providing valuable information for understanding the mechanical and fluid dynamic properties of the developing cardiac system.

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“…Hence, an intensity-modulated fluorescence signal at 1.7 MHz was detected in the experiment. 13 In the setup, a dichroic mirror separated the exciting laser beam and the fluorescent emission beam. The diameter of the pinhole aperture at 25 μm was chosen to match the objective lens O 1 (Olympus, LMPLFL 100X, NA ¼ 0.8, WD ¼ 3.4 mm) and the collector lens O 2 (Lightpath Gradium lenses, EFL ¼ 60 mm, f∕# ¼ 2.6).…”

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confidence: 99%

Improved axial point spread function in a two-frequency laser scanning confocal fluorescence microscope

Chung

Chien

et al. 2018

J. Biomed. Opt.

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A two-frequency laser scanning confocal fluorescence microscope (TF-LSCFM) based on intensity modulated fluorescence signal detection was proposed. The specimen-induced spherical aberration and scattering effect were suppressed intrinsically, and high image contrast was presented due to heterodyne interference. An improved axial point spread function in a TF-LSCFM compared with a conventional laser scanning confocal fluorescence microscope was demonstrated and discussed.

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Line-scan focal modulation microscopy

Cited by 15 publications

References 29 publications

Three‐dimensional cellular imaging in thick biological tissue with confocal detection of one‐photon fluorescence in the near‐infrared II window

Three‐dimensional cellular imaging in thick biological tissue with confocal detection of one‐photon fluorescence in the near‐infrared II window

Ultrahigh-speed line-scan SD-OCT for four-dimensional in vivo imaging of small animal models

Improved axial point spread function in a two-frequency laser scanning confocal fluorescence microscope

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