• Title/Summary/Keyword: fluorescence imaging

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A Multi-detection Fluorescence Dye with 5-ALA and ICG Using Modified Light Emitting Diodes

  • Yoon, Kicheol;Kim, Eunji;Kim, Kwanggi;Lee, Seunghoon;Yoo, Heon
    • Current Optics and Photonics
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    • v.3 no.3
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    • pp.256-262
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    • 2019
  • Extensive tumor resection accompanied by radiotherapy and chemotherapy is the standard of care for malignant gliomas. However, there is a significant obstacle to the complete resection of the tumor due to the difficulty of distinguishing tumor and normal brain tissue with a conventional surgical microscope. Recently, multiple studies have shown the possibility of fluorescence-guided surgery in malignant gliomas. The most used fluorescence dyes for brain tumor surgery are 5-aminolevulinic acid (5-ALA) and indocyanine green (ICG). In this paper, a new fluorescence guided operation system, which can detect both 5-ALA and ICG fluorescent images simultaneously, is presented. This operation system consists of light emitting diodes (LEDs) which emits 410 nm and 740 nm wavelengths. We have performed experiments on rats in order to verify the operation of the newly developed operation system. Oral administration and imaging were performed to observe the fluorescence of 5-ALA and ICG fluorescence in rats. When LEDs at wavelengths of 410 nm and 740 nm were irradiated on rats, 628 nm wavelength with a violet fluorescence color and 825 nm wavelength with a red fluorescence color were expressed in 5-ALA and ICG fluorescent material, respectively, thus we were able to distinguish the tumor tissues easily. Previously, due to the poor resolution of the conventional surgical microscope and the fact that the color of the vein is similar to that of the tumor, the tumor resection margin was not easy to observe, thus increasing the likelihood for cancer recurrence. However, when the tumor is observed through the fluorescence guided operation system, it is possible to easily distinguish the color with the naked eye and it can be completely removed. Therefore, it is expected that surgical removal of cancerous tumors will be possible and surgical applications and surgical microscopes for cancer tumor removal surgery will be promising in the future.

Development of Image-based System for Multiple Fluorescence Imaging Study (다중형광영상 연구를 위한 영상기반 시스템 개발)

  • Yoon, WoongBae;Kim, Hong Rae;Lee, Hyun Min;Kim, Young Jae;Kim, Kwang Gi;Yoo, Heon;Lee, Seung Hoon
    • Journal of Korea Multimedia Society
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    • v.18 no.12
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    • pp.1445-1452
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    • 2015
  • In these days, fluorescent materials such as ICG or 5-ALA is used for the brain surgery. The patients who underwent brain tumor surgery has been increased during last 30 years and the survivorship rate increased 22∼33% in 5 years. Recently, the Fluorescence induction surgery is developed for more safety and improved the resection rate for the glioma in the neurosurgery field. In this study, we proposed fluorescence area detection method for ICG and 5-ALA fluorescence induced surgery using acquired images from image processing. Accuracy was 99.21% from ICG images, and 99.51% from 5-ALA images. Matthews correlation coefficient was 88.67% from ICG images, and 90.49% from 5-ALA images.

Development of line-scanning two-photon microscopy based on spatial and temporal focusing for tryptophan based auto fluorescence imaging (고속 트립토판 자가형광 이미징을 위한 시공간적 집중 기반의 라인 스캐닝 이광자 현미경 개발)

  • Lee, Jun Ho;Nam, Hyo Seok;Kim, Ki Hean
    • Journal of the Korean Society of Visualization
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    • v.11 no.2
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    • pp.41-45
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    • 2013
  • Two-photon microscopy (TPM) is minimally-invasive 3D fluorescence microscopy based on nonlinear excitation, and TPM can visualize cellular structures based on auto-fluorescence. Line-scanning TPM is one of high-speed TPM methods without sacrificing the image resolution by using spatial and temporal focusing. In this paper, we developed line-scanning TPM based on spatial and temporal focusing for auto-fluorescence imaging by exciting the tryptophan. Laser source for this system was an optical parametric oscillator (OPO) and it made near 570 nm femtosecond pulse laser. It had 200fs pulse width and 1.72 nm bandwidth, so that the achievable depth resolution was 2.41um and field of view (FOV) is 10.8um. From the characterization, our system has 3.0 um depth resolution and 12.3 um FOV. We visualized fixed leukocyte cell sample and compared with point scanning system.

Application of black phosphorus nanodots to live cell imaging

  • Shin, Yong Cheol;Song, Su-Jin;Lee, Yu Bin;Kang, Moon Sung;Lee, Hyun Uk;Oh, Jin-Woo;Han, Dong-Wook
    • Biomaterials Research
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    • v.22 no.4
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    • pp.352-359
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    • 2018
  • Background: Black phosphorus (BP) has emerged as a novel class of nanomaterials owing to its unique optical and electronic properties. BP, a two-dimensional (2D) nanomaterial, is a structure where phosphorenes are stacked together in layers by van der Waals interactions. However, although BP nanodots have many advantages, their biosafety and biological effect have not yet been elucidated as compared to the other nanomaterials. Therefore, it is particularly important to assess the cytotoxicity of BP nanodots for exploring their potentials as novel biomaterials. Methods: BP nanodots were prepared by exfoliation with a modified ultrasonication-assisted solution method. The physicochemical properties of BP nanodots were characterized by transmission electron microscopy, dynamic light scattering, Raman spectroscopy, and X-ray diffractometry. In addition, the cytotoxicity of BP nanodots against C2C12 myoblasts was evaluated. Moreover, their cell imaging potential was investigated. Results: Herein, we concentrated on evaluating the cytotoxicity of BP nanodots and investigating their cell imaging potential. It was revealed that the BP nanodots were cytocompatible at a low concentration, although the cell viability was decreased with increasing BP nanodot concentration. Furthermore, our results demonstrated that the cells took up the BP nanodots, and the BP nanodots exhibited green fluorescence. Conclusions: In conclusion, our findings suggest that the BP nanodots have suitable biocompatibility, and are promising candidates as fluorescence probes for biomedical imaging applications.

Assessment and Comparison of Three Dimensional Exoscopes for Near-Infrared Fluorescence-Guided Surgery Using Second-Window Indocyanine-Green

  • Cho, Steve S.;Teng, Clare W.;Ravin, Emma De;Singh, Yash B.;Lee, John Y.K.
    • Journal of Korean Neurosurgical Society
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    • v.65 no.4
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    • pp.572-581
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    • 2022
  • Objective : Compared to microscopes, exoscopes have advantages in field-depth, ergonomics, and educational value. Exoscopes are especially well-poised for adaptation into fluorescence-guided surgery (FGS) due to their excitation source, light path, and image processing capabilities. We evaluated the feasibility of near-infrared FGS using a 3-dimensional (3D), 4 K exoscope with near-infrared fluorescence imaging capability. We then compared it to the most sensitive, commercially-available near-infrared exoscope system (3D and 960 p). In-vitro and intraoperative comparisons were performed. Methods : Serial dilutions of indocyanine-green (1-2000 ㎍/mL) were imaged with the 3D, 4 K Olympus Orbeye (system 1) and the 3D, 960 p VisionSense Iridium (system 2). Near-infrared sensitivity was calculated using signal-to-background ratios (SBRs). In addition, three patients with brain tumors were administered indocyanine-green and imaged with system 1, with two also imaged with system 2 for comparison. Results : Systems 1 and 2 detected near-infrared fluorescence from indocyanine green concentrations of >250 ㎍/L and >31.3 ㎍/L, respectively. Intraoperatively, system 1 visualized strong near-infrared fluorescence from two, strongly gadolinium-enhancing meningiomas (SBR=2.4, 1.7). The high-resolution, bright images were sufficient for the surgeon to appreciate the underlying anatomy in the near-infrared mode. However, system 1 was not able to visualize fluorescence from a weakly-enhancing intraparenchymal metastasis. In contrast, system 2 successfully visualized both the meningioma and the metastasis but lacked high resolution stereopsis. Conclusion : Three-dimensional exoscope systems provide an alternative visualization platform for both standard microsurgery and near-infrared fluorescent guided surgery. However, when tumor fluorescence is weak (i.e., low fluorophore uptake, deep tumors), highly sensitive near-infrared visualization systems may be required.

Optimal Optical Filters of Fluorescence Excitation and Emission for Poultry Fecal Detection

  • Kim, Tae-Min;Lee, Hoon-Soo;Kim, Moon-S.;Lee, Wang-Hee;Cho, Byoung-Kwan
    • Journal of Biosystems Engineering
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    • v.37 no.4
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    • pp.265-270
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    • 2012
  • Purpose: An analytic method to design excitation and emission filters of a multispectral fluorescence imaging system is proposed and was demonstrated in an application to poultry fecal inspection Methods: A mathematical model of a multispectral imaging system is proposed and its system parameters, such as excitation and emission filters, were optimally determined by linear discriminant analysis (LDA). An alternating scheme was proposed for numerical implementation. Fluorescence characteristics of organic materials and feces of poultry carcasses are analyzed by LDA to design the optimal excitation and emission filters for poultry fecal inspection. Results: The most appropriate excitation filter was UV-A (about 360 nm) and blue light source (about 460 nm) and band-pass filter was 660-670 nm. The classification accuracy and false positive are 98.4% and 2.5%, respectively. Conclusions: The proposed method is applicable to other agricultural products which are distinguishable by their spectral properties.

Research about Hyperspectral Imaging System for Pre-Clinical testing of Small Animal (소형동물 전임상실험을 위한 하이퍼스펙트럼 영상장비 연구)

  • Lee, kyeong-Hee;Choi, Young-Wook
    • The Transactions of The Korean Institute of Electrical Engineers
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    • v.56 no.12
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    • pp.2208-2213
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    • 2007
  • In this study we have developed a hyperspectrum imaging system for highly sensitive and effective imaging analysis. An optical setup was designed using acoustic optical tunable filter (AOTF) for high sensitive hyperspectrum imaging. Light emitted by mercury lamp gets split in to diffracted and undiffracted beams while passing though AOTF. GFP transfected HEK-293 cell line was used as a model for in vitro imaging analysis. Cells were first, analyzed by fluorescence microscope followed by flow cytometric analysis. Flow cytometric analysis showed 66.31% transfection yield in GFP transfected HEK-293 cells. Various images of GFP transfected HEK-293 cell were grabbed by collecting the diffracted light using a CCD over a dynamic range of frequency of 129-171 MHz with an interval of 3 MHz. Subsequently, for in vivo image analysis of GFP transfected cells in mouse, a whole-body-imaging system was constructed. The blue light of 488 nm wavelength was obtained from a Xenon arc lamp using an appropriate filter and transmitted through an optical cable to a ring illuminator. To check the efficacy of the newly developed whole-body-imaging system, a comparative imaging analysis was performed on a normal mouse in presence and absence of Xenon arc irradiation. The developed hyperspectrum imaging analysis with AOTF showed the highest intensity of green fluorescent protein at 153 MHz of frequency and 494 nm of wavelength. However, the fluorescence intensity remained same as that of the background below 138 MHz (475 nm) and above 162 MHz (532 nm). The mouse images captured using the constructed whole-body-imaging system appeared monochromatic in absence of Xenon arc irradiation and blue when irradiated with Xenon arc lamp. Nevertheless, in either case mouse images appeared clearly.

A Study on Characteristics Analysis of Multichannel Filter Module for Near-infrared Fluorescence Imaging (근적외선 형광 이미징 영상 구현을 위한 다채널 필터 모듈 특성분석 연구)

  • Choi, Jinsoo;Cho, Sang Uk;Kim, Doo-In;Lee, Hak-Guen;Choi, Hak Soo;Jeong, Myung Yung
    • Journal of the Microelectronics and Packaging Society
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    • v.23 no.1
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    • pp.29-34
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    • 2016
  • In this study, development of multichannel filter module and characteristic evaluation for bio imaging were studied. The filter module was fabricated in order to realize near infrared fluorescence imaging of 700 nm and 800 nm wavelength ranges, and contrast imaging analysis for characteristic evaluation of the filter module was studied through signal to back ground ratio (SBR), controlled by parameters such as magnification, exposure, gain. Furthermore, phantoms, which are biomimetic tissue with equal optical properties of kidney and liver, were fabricated to study characteristics of both filter module depending on thickness and exposure amount of light source for bio imaging analysis. The fabricated filter module has more than 4 of SBR difference despite changes of magnification, exposure, gain, and in the case of the kidney phantom and the liver phantom, contrast imaging of more than 4 of SBR was confirmed on 50 mA, 60 mA exposure amount of light source respectively.

Study on Fluorescence Excitation and Emission Bands for Detection of Fecal Residue on Poultry Carcasses (도계육 표면 내장 잔유물 오염 검사를 위한 형광 조사 및 방출파장 선정에 관한 연구)

  • Cho, B.K.;Kim, M.S.
    • Journal of Biosystems Engineering
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    • v.33 no.2
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    • pp.136-141
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    • 2008
  • Fecal contaminations on poultry carcasses, not easily discemable by human eyes, are potential harbor sites of pathogenic Escherichia Coli (E. coli O157:H7). Development of sensitive detection methods for fecal contamination is essential to ensure safe production of poultry products. Fluorescence has been shown to be very sensitive in detecting fecal and other biological substances that can harbor pathogens. In this study, fluorescence excitation-emission spectra of poultry fecal matter were compared with spectra for poultry skin and meat. Results indicated that the combinations of fluorescence intensities at the wavelength of 520 nm, 579 nm, 625 nm, and 635 nm with 411 nm excitation showed above 97% accuracy for differentiation of the contaminants from poultry tissues. Excitation and emission bands determined could be used for constructing a real-time fluorescence imaging device for detection of harmful residues on poultry carcasses.

Development and Evaluation of Multi-Wavelength Excitation light Source for Fluorescence Imaging to Diagnose Malignancies (악성종양의 형광영상 진단을 위한 다파장 여기광원장치의 개발과 평가)

  • Lim, Hyun-Soo
    • Journal of Biomedical Engineering Research
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    • v.30 no.2
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    • pp.113-121
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    • 2009
  • This study aims at designing and evaluating light source devices that can stably generate light with various wavelengths in order to make possible PDD using a photosensitizer and diagnosis using auto-fluorescence. The light source was a Xenon lamp and filter wheel, composed of an optical output control through Iris and filters with several wavelength bands. It also makes the inducement of auto-fluorescence possible because it is designed to generate a wavelength band of 380-420nm, 430-480nm, and 480-560nm. The transmission part of the light source was developed to enhance the efficiency of light transmission. To evaluate this light source, the characteristics of light output and wavelength band were verified. To validate the capability of this device as PDD, the detection of auto-fluorescence using mouse models was performed.