Earticle

Over the past 40 years, there was significant increase in research towards the field of molecular electronic to overcome the limitations of silicon based devices. Among several types of molecular devices, molecular memory using biomaterial is a major research theme nowadays. The naturally occurring biomaterials like metalloproteins are nano-sizes and possess inherently charge retention property, which make them to use as a high density molecular device platform. Pseudomonas aeruginosa azurin, the protein of interest in this research, has copper as a key element in the electron transfer mechanisms. This can be used as an electron acceptor in the development of molecular electronic device by mimicking biological mechanism. To achieve their information storage properties on solid state with reproducibility and reliability, the robust fabrication of molecular layers with fine orientation are one of the key processes. We have modified azurin with different number of cysteine residues at its amino acid chain based on site-directed mutagenesis. The resulting recombinant protein, azurin, retained its original redox property in the same manner as native azurin. Recombinant azurin was immobilized on Au substrate by strong affinity between thiol of cysteine and gold. The orientations of recombinant azurin with cystein residues immobilized on the Au substrate were analyzed by fluorescence microscope, scanning tunneling microscope, and surface plasmon resonance. Our data revealed that binding activity of recombinant azurin with two cystein residue on the Au substrate significantly increased in comparison to single residue auzrin. Immobilization of highly oriented recombinant azurin based on cysteine-modification could be useful for the nanoscale fabrication of bioelectronic device. Acknowledgments: This research was supported by the Nano/Bio Science & Technology Program of the Ministry of Education, Science and Technology (MEST), by the Korea Science and Engineering Foundation (KOSEF) grant funded by the Korea government (MEST) (2006-05374), and by Seoul Research and Business Development Program (10816).

We report the new threadlike structures outside the blood vessels of mice. For this, we developed an in-situ searching method of the structure by vital staining with the dye of acridine orange and using a fluorescent stereomicroscope designed specifically for this purpose. We consider that the newly found threadlike structure might be rediscovery of the extra-vascular Bonghan duct which was reported in 1963 by Bonghan Kim.

In this study, we simulated a statistical model of FCS(fluorescence correlation spectroscopy) based on a Poisson process to understand and explain observations of the experiment performed on molecules of ultra-low concentration by the home-built laser-scanning confocal microscope. The statistical model confirmed that the relative mean square amplitude of fluctuations is shown to be inversely proportional to the average number of molecules, even in the ultra-low concentration, if some conditions are satisfied. Signal-to-noise ratio and the variability of dwelling time under the confocal volume were found to be effective conditions for the experiment.

In order to analyze cell images, accurate segmentation of each cell is indispensable. However, the reality is that accurate cell image segmentation is not easy due to various noises, dense cells, and inconsistent shape of cells. Therefore, in this paper, we propose an algorithm that combines marker-based watershed segmentation and ellipse fitting method for glioblastoma cell segmentation. In the proposed algorithm, in order to solve the over-segmentation problem of the existing watershed method, the marker-based watershed technique is primarily performed through "seeding using local minima". In addition, as a second process, the concave point search using ellipse fitting for final segmentation based on the connection line between the concave points has been performed. To evaluate the performance of the proposed algorithm, we compared three algorithms with other algorithms along with the calculation of segmentation accuracy, and we applied the algorithm to other cell image data to check the generalization and propose a solution.

본 연구에서는 뇌 종양 수술에서 다수의 광원과 빔 스플리터 모듈을 사용해 종양과 혈관의 형광영상을 동시에 검출하고 획득한 형광영상을 동일한 디스플레이 장치에 표시함으로써 시술자에게 종양과 혈관의 정확한 정보를 실시간으로 제공할 수 있는 현미경 시스템을 제안한다. 5-ALA(5-Aminolevulinic acid) 와 ICG(Indocyanine green) 의 형광영상의 동시 검출을 위해 빔 스플리터(beam-splitter : BS)모듈을 사용하였고 5-ALA는 600nm, ICG는 800nm이상의 파장 대역에서 가장 효율이 뛰어나도록 구성하였다. 빔 스플리터 모듈은 파장 대역에 따라 광학기기의 구조를 변경할 수 있고 필터를 탈, 착 가능한 구조로 설계하여 필요에 따라 빔 스플리터와 필터의 종류를 변경할 수 있으며 5-ALA 및 ICG 이외의 형광염료를 사용한 시술에서 사용할 수 있다. 빔 스플리터 모듈을 통한 형광영상은 5-ALA는 가시광역, ICG는 근적외선 영역을 검출 할 수 있는 CCD 카메라를 장착해 동일한 디스플레이에서 확인할 수 있고 획득한 형광영상은 닮음 변환(similarity transform)을 이용해 원영상과 정합하여 실시간으로 시술자에게 제공하는 시스템을 구현하였다.

In this paper, we propose a microscope system for detecting both a tumor and blood vessels in brain tumor surgery as fluorescence images by using multiple light sources and a beam-splitter module. The proposed method displays fluorescent images of the tumor and blood vessels on the same display device and also provides accurate information about them to the operator. To acquire a fluorescence image, we utilized 5-ALA (5-aminolevulinic acid) for the tumor and ICG (Indocyanine green) for blood vessels, and we used a beam-splitter module combined with a microscope for simultaneous detection of both. The beam-splitter module showed the best performance at 600 nm for 5-ALA and above 800 nm for ICG. The beam-splitter is flexible to enable diverse objective setups and designed to mount a filter easily, so beam-splitter and filter can be changed as needed, and other fluorescent dyes besides 5-ALA and ICG are available. The fluorescent images of the tumor and the blood vessels can be displayed on the same monitor through the beam-splitter module with a CCD camera. For ICG, a CCD that can detect the near-infrared region is needed. This system provides the acquired fluorescent image to an operator in real time, matching it to the original image through a similarity transform.