Mr. Hyeong-Jun Jeong Profile

Hyeong-Jun Jeong

at KAIST

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Proceedings Article | 22 February 2013 Paper

Dual detection confocal fluorescence microscopy: depth imaging without depth scanning

Dong-Ryoung Lee, Young-Duk Kim, Hyeong-Jun Jeong, Jong-Min Lee, Dae-Gab Gweon, Hongki Yoo

Proceedings Volume 8589, 85891G (2013) https://doi.org/10.1117/12.2003005

KEYWORDS: Confocal microscopy, Luminescence, Microscopy, Quantum efficiency, Fluorescence spectroscopy, 3D image processing, Mirrors, Beam splitters, Ferroelectric materials, 3D scanning

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We propose a new method for three-dimensional fluorescence imaging without depth scanning that we refer to as the dual detection confocal fluorescence microscopy (DDCFM). Compared to conventional beam-scanning confocal fluorescence microscopy, where the focal spot must be scanned either optically or mechanically to collect a three-dimensional images, DDCFM is able to obtain depth information without depth scanning. DDCFM utilizes two photo multiplier tubes (PMTs) in the confocal detection system. The emitted fluorescence is divided by the beam splitter and received by the two PMTs through pinholes with different size. Each PMT signal generates different axial response curve according to the pinhole diameter, which decides stiffness of the curve. Since the PMT signal is determined by the intensity of the fluorescent emitter and the distance from the focal point, we can acquire depth position of a fluorescent emitter by comparing two intensity signals from the PMTs. Since the depth information can be obtained by a single excitation without depth scanning, DDCFM has many advantages. The measurement time is dramatically reduced for volume imaging. Also, photo-bleaching and photo-toxicity can be minimized. The system can be easily miniaturized because no mechanical depth scan is needed. Here, we demonstrate the feasibility of the proposed method by phantom study using fluorescent beads.

Proceedings Article | 23 February 2011 Paper

Design and analysis of confocal-spectral microscopy using wavelength scanning scheme

Dukho Do, Wanhee Chun, Hyeongjun Jeong, Dae-Gab Gweon

Proceedings Volume 7904, 79041L (2011) https://doi.org/10.1117/12.873667

KEYWORDS: Sensors, Luminescence, Microscopy, Mirrors, Signal detection, Calcite, Birefringence, Confocal microscopy, Polarization, Fluorescence spectroscopy

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The multi-color, or spectral fluorescence microscopy has ability to detect fluorescence spectral signals which are useful in case of studying interactions and phenomena between biological samples. Recently, commercial devices are combining with confocal microscope so to enhance lateral resolution and to have axial direction discernment. Also Acousto-Optic Tunable Filter(AOTF) is used instead of dichroic mirror to divide excitation and emission signals with mininum light efficiency. In addition, AOTF is used in spectral fluorescence microscopy have many advantages, these are very fast switching speed and high resolution in wavelength selection. However it uses acousto-optic interactions in birefringence material, Tellurium Dioxide(TeO₂), the excitation light interacts with appropriate acoustic signal so that it is diffracted to 1 or -1 order path. But the fluorescence signals from a sample propagate in 0 order path with small different angle according to the polarization state. In this paper, a confocal-spectral microscopy is proposed with the new kind of spectral detector design having wavelength scanning galvano mirror. It makes possible to detect broad wavelength fluorescence signal by single PMT with simply rotating the galvano mirror. Also a new birefringent material, calcite(CaCO₃) is used to compensate polarization effect. The proposed spectral confocal microscopy with unique spectrometer body has many advantages in comparison with commercial devices. In terms of detection method, it can be easily applied to other imaging modalities. Hence this system will be adapted in many applications.

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