• Applied Microscopy
• /
• 제51권
• /
• pp.2.1-2.7
• /
• 2021

Synaptic vesicles, which are endogenous to neurotransmitters, are involved in exocytosis by active potentials and release neurotransmitters. Synaptic vesicles used in neurotransmitter release are reused via endocytosis to maintain a pool of synaptic vesicles. Synaptic vesicles show different types of exo- and endocytosis depending on animal species, type of nerve cell, and electrical activity. To accurately understand the dynamics of synaptic vesicles, direct observation of synaptic vesicles is required; however, it was difficult to observe synaptic vesicles of size 40-50 nm in living neurons. The exo-and endocytosis of synaptic vesicles was confirmed by labeling the vesicles with a fluorescent agent and measuring the changes in fluorescence intensity. To date, various methods of labeling synaptic vesicles have been proposed, and each method has its own characteristics, strength, and drawbacks. In this study, we introduce methods that can measure presynaptic activity and describe the characteristics of each technique.

• Journal of Microbiology
• /
• 제40권4호
• /
• pp.319-326
• /
• 2002

Cellular DNA in prokaryotes is organized in nucleic acid-protein self-assemblies referred to as the nucleoid. The physical forces responsible for its stability inside the poor solvent properties of the cytoplasm and their functional implications are not understood. Studies on the organisation and functioning of the cytosol of cells largely rely on experimental protocols performed in highly dilute solutions using biochemically purified molecules, which is not a reliable substitute for the situation existing in vivo. Our current research interest is focused on the characterization of biological and physical forces determining the compaction and phase separation of DNA in Escherichia coli cytoplasm. We have emphasized the effect of excluded volume in solutions with high macromolecular concentrations (macromolecular crowding) upon self-association patterns of reactions. The prokaryotic cytosol was simulated by addition of inert polymer polyethylene glycol (PEG) (average molecular weight 20000), as an agent which afterwards facilitates the self-association of macromolecules. Fluorescence microscopy was used for direct visualization of nucleoids in intact cells, after staining with DAPI (4',6-diamidino-2-phenylindole dihydrochloride). Addition of the crowding agent PEG 20,000, in increasing concentrations generated progressively enhanced nucleoid compaction, the effect being stronger in the presence of 0.2 M NaCl and 5 mM MgCl$\_$2/. Under these conditions, the nucleoids were compacted to volumes of around 2 ㎛$\^$3/ or comparable sizes with that of living cells.

• 한국산림과학회지
• /
• 제67권1호
• /
• pp.28-30
• /
• 1984

형광색소(蛍光色素) DAPI(4'-6-diamidino-2-phenylindole, 2HCI)와 형광원미경(蛍光願微鏡)을 이용(利用)한 조직화학적(組織化學的) 기법(技法)으로 빗자루병(病)에 걸린 대추나무 조직내(組織內)의 마이코플라스마 분포(分布)를 조사(調査)한 결과(結果), 빗자루병징(病徵)이 나타나 있는 가지에서는 외관상(外觀上) 건전엽(健全葉)을 포함(包含)하여 모든 잎과 줄기에서 마이코 플라스마가 검출(檢出)되었으나 병징(病徵)이 나타나 있지 않은 가지의 잎과 줄기에서는 마이코플라스마가 검출(檢出)되지 않았다. 또한 감염(感染)된 나무의 뿌리조직(組織)에서도 뚜렷한 형광반응(蛍光反應)이 나타남으로써 마이코플라스마가 뿌리조직(組織)에도 많이 존재하고 있음을 알 수 있다.

• The Plant Pathology Journal
• /
• 제17권6호
• /
• pp.329-335
• /
• 2001

Yellowing disease of palms caused by phytoplasma is spreading in the Arabian Gulf region. Surveys were conducted to determine the occurrence of the disease. Electron and fluorescence microscopy, and polymerase chain reaction (PCR) techniques were used to detect the phytoplasma associated with the yellowing disease of ornamental palm Washingtonia sp. grown in Kuwait. An accumulation of phytoplasmal DNA was observed by fluorescence microscopy in phloem tissues of diseased palms. Electron microscopy showed that phytoplasma cells were primarily confined to the phloemsieve elements of tissue samples collected from infected mature palms in the field. The pathogen was identified on the basis of molecular analysis using universal and specific nested primers in PCR amplifications. Prokaryotic 16S rDNA gene was detected in amplified PCR products. Nested PCR resulted in DNA amplification of 1.2 kbp fragment. This is the first report of a phytoplasmal rDNA gene identified from the putative causal pathogen of yellows in ornamental palms in the Arabian Gulf region.

• 한국광과학회:학술대회논문집
• /
• 한국광과학회 2002년도 정기총회 및 제9회 광과학 학술심포지움 논문초록
• /
• pp.32-32
• /
• 2002

• Molecules and Cells
• /
• 제38권11호
• /
• pp.975-981
• /
• 2015

Precise 3D spatial mapping of cells and their connections within living tissues is required to fully understand developmental processes and neural activities. Zebrafish embryos are relatively small and optically transparent, making them the vertebrate model of choice for live in vivo imaging. However, embryonic brains cannot be imaged in their entirety by confocal or two-photon microscopy due to limitations in optical range and scanning speed. Here, we use light-sheet fluorescence microscopy to overcome these limitations and image the entire head of live transgenic zebrafish embryos. We simultaneously imaged cranial neurons and blood vessels during embryogenesis, generating comprehensive 3D maps that provide insight into the coordinated morphogenesis of the nervous system and vasculature during early development. In addition, blood cells circulating through the entire head, vagal and cardiac vasculature were also visualized at high resolution in a 3D movie. These data provide the foundation for the construction of a complete 4D atlas of zebrafish embryogenesis and neural activity.

• Restorative Dentistry and Endodontics
• /
• 제26권1호
• /
• pp.1-8
• /
• 2001

Dental Caries which has high prevalence rate, accounts for majority of dental diseases. Many treatment and preventive treatment has been developed, thereby reducing the prevalence rate, but in our country, fluoridization has not spread widely yet, so prevention has not been done satisfactorily. When dental caries progresses, irreversible damage of tooth structure occurs. In initial dental caries, demineralizing tooth structure can be remineralized, so restorative treatment is unnecessary. In this study, 20 teeth restored with composite resin without fluoride release were used and divided into two groups. Incipient dental caries were artificially made and demineralization procedure was done for 1 and 2 weeks, for each group. Changes in mineral contents around the margins were analysed with confocal laser scanning microscope. The results were as follow. 1. Both total fluorescence of the lesion and average fluorescence of the lesion of remineralized samples decreased compared to demineralizing state. (p＜0.01) 2. Confocal laser scanning microscopy can be used in quantitative analysis of mineral change. In result, confocal laser scanning microscopy can be used in quantitative analysis of mineral change and it could be used in many different fields of dentistry in the future.

• 정보저장시스템학회논문집
• /
• 제11권2호
• /
• pp.22-25
• /
• 2015

In this paper, we demonstrate solid-immersion lens based confocal microscopy using super-continuum generation effect. Using super-continuum generation effect, we could diversify the excitation wavelength of confocal microscopy. Further, high refractive index of solid-immersion lens would increase the resolution of confocal microscopy. As a result, by applying the super-continuum generation effect and solid-immersion lens to confocal microscopy, some problems of confocal fluorescent microscopy, the excitation wavelength and the resolution, could be overcome. To verify it, we made home-built solid-immersion lens based confocal microscopy using super-continuum generation effect, and evaluate the performance of the system.

• Journal of the Optical Society of Korea
• /
• 제16권1호
• /
• pp.42-46
• /
• 2012

We have developed a fluorescence optical detection system using a digital micromirror device (DMD) for monitoring 3D cell culture matrices in situ. Full 3D imaging with fast scanning speed was implemented by the combined action of a DMD and a motorized stage. Imaging results with fluorescent microbeads measure the minimum axial resolution of the system as $6.3{\mu}m$, while full 1-mm scanning through 3D alginate-based matrix was demonstrated. For cell imaging, improved images were obtained by removing background fluorescence although the scanning distance was reduced because of low intracellular fluorescence efficiency. The system is expected to be useful to study various dynamics and behaviors of 3-dimensionally cultured cells in microfluidic systems.

• Bulletin of the Korean Chemical Society
• /
• 제34권9호
• /
• pp.2725-2730
• /
• 2013

Single C-reactive protein (CRP) molecules, which are non-specific acute phase markers and products of the innate immune system, were quantitatively detected on a gold-nanopatterned biochip using evanescent field-enhanced fluorescence imaging. The $4{\times}5$ gold-nanopatterned biochip (spot diameter of 500 nm) was fabricated by electron beam nanolithography. Unlabeled CRP molecules in human serum were identified with single-molecule sandwich immunoassay by detecting secondary fluorescence generated by total internal reflection fluorescence (TIRF) microscopy. With decreased standard CRP concentrations, relative fluorescence intensities reduced in the range of 33.3 zM-800 pM. To enhance fluorescence intensities in TIRF images, the distance between biochip surface and CRP molecules was optimally adjusted by considering the quenching effect of gold and the evanescent field intensity. As a result, TIRF only detected one single-CRP molecule on the biochip the first time.