• Title/Summary/Keyword: single cell imaging

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Molecular Structure of Muscle Filaments Determined by Electron Microscopy

  • Craig, Roger
    • Applied Microscopy
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    • v.47 no.4
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    • pp.226-232
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    • 2017
  • Electron microscopy and X-ray diffraction have together played a key role in our understanding of the molecular structure and mechanism of contraction of muscle. This review highlights the role of electron microscopy, from early insights into thick and thin filament structure by negative staining, to studies of single myosin molecule structure, and finally to recent high-resolution structures by cryo-electron microscopy. Muscle filaments are designed for movement. Their labile structures thus present challenges to obtaining near-atomic detail, which are also discussed.

Common-path phase microscopy for lives cell imaging (살아있는 세포 영상획득을 위한 common-path phase microscopy)

  • Lee, Ji-Yong;Lee, Seung-Rak;Yang, W.Z.;Kim, Deok-Yeong
    • Proceedings of the Optical Society of Korea Conference
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    • 2008.07a
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    • pp.273-274
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    • 2008
  • We present a quantitative phase microscopy for live cells. This method uses the principles of common path inteferometry and single shot phase image. This system has the ability to measure live cells quantitatively with subnanometer path length stability and millisecond scale aquisition time.

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Production of the Recombinant Single Chain Anti-B Cell Lymphoma Antibody and Evaluation of Immunoreactivity (pET vector를 통한 유전자 재조합 단일사슬 항 B형 림프종 항체의 생산과 면역반응성 평가)

  • Jung, Jae-Ho;Choi, Tae-Hyun;Woo, Kang-Sun;Chung, Wee-Sup;Kim, Soo-Gwan;Cheon, Gi-Jeong;Choi, Chang-Woon;Lim, Sang-Moo
    • Nuclear Medicine and Molecular Imaging
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    • v.40 no.4
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    • pp.211-217
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    • 2006
  • Purpose: Recombinant ScFv lym-1 was produced, using pET vector system for large scale production. Methods: ScFv lym-1 gene inserted pET-22b (+) vector, was expressed in E.coli BL-21 strain. ScFv lym-1 antibody extracted from periplasm, was purified with His-Taq column. To evaluated immunoreactivity with Raji cell, ScFv lym-1 was labeled with I-125 and I-125 ScFv lym-1 was purified with desalting column. Raji cell was injected into the C57BR/cdJ SCID mice. Gamma camera imaging were taken time point at 1, 8, 24, and 48 hr with 8 mm pinhole collimator. Results: An active scFv lym-1 could be produced in E. coli with soluble iron using PET vector system. Immuuoreaetivity and affinity constant of IgG lym-1 were 54% and $1.83{\times}10^9M^{-1}$, respectively, and those of scFv lym-1 were 53.7% and $1.46{\times}10^9M^{-1}$, respectively. Biodistribution of I-125 scFv lym-1 antibody showed faster clearance in blood, spleen, kidney and than I-125 IgG lym-1 antibody. Gamma camera image of I-125 scFv lym-1 antibody showed faster clearance and tumor targeting liver than I-125 IgG lym-1 antibody. Conclusions: In vitro properties of scFv lym-1 were similar to those of IgG lym-1. ScFv lym-1 showed faster blood clearance than IgG lym-1 There results suggest that scFv lym-1 antibody can be useful for tumor imaging agent.

A comparison of photosynthesis measurements by O2 evolution, 14C assimilation, and variable chlorophyll fluorescence during light acclimatization of the diatom Coscinodiscus granii

  • Trampe, Erik;Hansen, Per Juel;Kuhl, Michael
    • ALGAE
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    • v.30 no.2
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    • pp.103-119
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    • 2015
  • Photosynthetic rates of the large centric diatom Coscinodiscus granii were measured by means of multicolor variable chlorophyll fluorescence imaging, single cell $^{14}C$ assimilation, and optical $O_2$ sensor measurements during light acclimatization of cultures grown at five different irradiances: 50, 150, 235, 332, and $450{\mu}mol$ photons $m^{-2}\;s^{-1}$. Photo-acclimatization was evident from changes of cellular chlorophyll a content, growth rates, and light response curves. Each of the applied methods evaluates different parts and reactions in the photosynthetic apparatus, which makes a direct quantitative comparison of rates difficult, although a different degree of correlation were found between all three methods. However, when used in combination, they provide information about the internal relationship of photosynthetic pathways as well as the variation in photosynthetic capacity between individual cells within a single algal culture.

Non-invasive evaluation of embryo quality for the selection of transferable embryos in human in vitro fertilization-embryo transfer

  • Jihyun Kim;Jaewang Lee;Jin Hyun Jun
    • Clinical and Experimental Reproductive Medicine
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    • v.49 no.4
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    • pp.225-238
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    • 2022
  • The ultimate goal of human assisted reproductive technology is to achieve a healthy pregnancy and birth, ideally from the selection and transfer of a single competent embryo. Recently, techniques for efficiently evaluating the state and quality of preimplantation embryos using time-lapse imaging systems have been applied. Artificial intelligence programs based on deep learning technology and big data analysis of time-lapse monitoring system during in vitro culture of preimplantation embryos have also been rapidly developed. In addition, several molecular markers of the secretome have been successfully analyzed in spent embryo culture media, which could easily be obtained during in vitro embryo culture. It is also possible to analyze small amounts of cell-free nucleic acids, mitochondrial nucleic acids, miRNA, and long non-coding RNA derived from embryos using real-time polymerase chain reaction (PCR) or digital PCR, as well as next-generation sequencing. Various efforts are being made to use non-invasive evaluation of embryo quality (NiEEQ) to select the embryo with the best developmental competence. However, each NiEEQ method has some limitations that should be evaluated case by case. Therefore, an integrated analysis strategy fusing several NiEEQ methods should be urgently developed and confirmed by proper clinical trials.

Effects of irradiation on the mRNA expression of the osteocalcin and osteopontin in MC3T3-E1 osteoblastic cell line (MC3T3-E1 조골세포주의 osteocalcin과 osteopontin mRNA 발현에 미치는 방사선의 영향)

  • Cho Su-Beom;Lee Sang-Rae;Koh Kwang-Joon
    • Imaging Science in Dentistry
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    • v.33 no.3
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    • pp.179-185
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    • 2003
  • Purpose: To investigate the effects of irradiation on the phenotypic expression of the MC3T3-El osteoblastic cell line, particularly on the expression of osteocalcin and osteopontin. Materials and Methods: Cells were irradiated with a single dose of 0.5, 1,4, and 8 Gy at a dose rate of 5.38 Gy/min using a cesium 137 irradiator. After the specimens were harvested, RNA was extracted on the 3rd, 7th, 14th, and 21st day after irradiation. The RNA strands were reverse-transcribed and the resulting cDNAs were subjected to amplification by PCR. Results: The irradiated cells demonstrated a dose-dependent increase in osteocalcin and a dose-dependent decrease in osteopontin mRNA expression compared with the non-irradiated control group, The amount of osteocalcin mRNA expression decreased significantly at the 3rd day after irradiation of 0,5, 1,4, and 8 Gy, and also decreased significantly at the 3rd, 14th, and 21 st day after irradiation in the 8 Gy exposed group compared with the control group, The degree of osteopontin mRNA expression increased significantly at the 7th day after irradiation of 0,5, 1,4, and 8Gy, Conclusion: These results showed that each single dose of 0,5, 1, 4, and 8 Gy influenced the mRNA expression of osteocalcin and osteopontin associated with the calcification stage of osteoblastic cells, suggesting that each single dose affected bone formation at the cell level.

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Diagnostic imaging of portosystemic shunts in 43 dogs (개 문맥전신순환단락의 진단영상 43례)

  • Choi, Jihye;Kim, Hyunwook;Jang, Jaeyoung;Kim, Junyoung;Yoon, Junghee
    • Korean Journal of Veterinary Research
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    • v.48 no.2
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    • pp.227-233
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    • 2008
  • Portosystemic shunt (PSS) was diagnosed in 43 dogs by mesenteric portogram from January, 2002 to June 2007 in Haemaru referral animal hospital. PSS was found in various breeds including Maltese, Miniature Schnauzer and Yorkshire Terrier and there was no predisposition in gender. In laboratory parameters, mean cell volume was lower than normal value in single shunt and alanine aminotransferase was higher than normal range in multiple shunts with clinical significance. Cystic calculi were found in over 50% dogs with PSS and even in 70.8% dogs with single shunt. In 81% dogs with PSS, extrahepatic single shunt such as portocarval type and portoazygous type was identified. Extrahepatic multiple shunt and intrahepatic single shunt were observed in 4 dogs, respectively. Gradual attenuation using ameroid constrictor was applied to 35 dogs with extrahepatic single shunt and the prognosis of these dogs were good except two dogs, which showed poor prognosis because of acquired multiple PSS and renal disease unrelated with PSS, respectively.

Carbon Nanotubes Multi Electrodes Array to Image Capacitance for Label-free Discrimination of Lipid Region in Atherosclerosis ex vivo

  • Song, Jun-Ho;Lee, Seon-Mi;Han, Nal-Ae;Yu, Gyeong-Hwa
    • Proceedings of the Korean Vacuum Society Conference
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    • 2016.02a
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    • pp.372.1-372.1
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    • 2016
  • Recently, there are a lot of diseases all around the world. Out of them, Atherosclerosis (AS) is the most common cause of stroke, cardiovascular mortality, and myocardial infarction. The macrophage-derived foam cell, which is formed by oxidized low-density lipoprotein (oxLDL), is the crucial marker for AS. In this study, we report a label-free capacitance imaging technique with multi-electrode array (MEA). The lipid-rich aorta arch lesions, which are derived from an apolipoprotein-E receptor-deficient (apoE-/-) mouse, exhibit higher capacitance than the lipid-free aorta arch, allowing the capacitance imaging of lipid region in atherosclerosis. To improve the contacts between MEA and tissue, polypyrrole(PPy)-coated multi walled carbon nanotubes (MWNTs) multi electrode array (PPy-MWNTs-MEA) was fabricated. Compared to TiN-MEA, PPy-MWNTs-MEA yielded lower contact impedance and better capacitance images. In addition, we have also developed a flexible MEA using single walled carbon nanotubes on a PET substrate. The lipid region could be discriminated in the capacitance images of the lipid-rich aorta arch lesions measured using flexible MEA, demonstrating a feasibility of in vivo applications.

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Visualization of Epidermis and Dermal Cells in ex vivo Human Skin Using the Confocal and Two-photon Microscopy

  • Choi, Sang-Hoon;Kim, Wi-Han;Lee, Yong-Joong;Lee, Ho;Lee, Weon-Ju;Yang, Jung-Dug;Shim, Jong-Won;Kim, Jin-Woong
    • Journal of the Optical Society of Korea
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    • v.15 no.1
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    • pp.61-67
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    • 2011
  • The confocal laser scanning microscopy and two-photon microscopy was implemented based on a single laser source and an objective lens. We imaged and compared the morphology of identical sites of ex vivo human skin using both microscopes. The back-scattering emission from the sample provided the contrast for the confocal microscopy. The intrinsic autofluorescence and the second harmonic generation were used as the luminescence source for the two-photon microscopy. The wavelength of the Ti:Sapphire laser was tuned at 710 nm, which corresponds to the excitation peak of NADH and FAD in skin tissue. The various cell layers in the epidermis and the papillary dermis were clearly distinguished by both imaging modalities. The two-photon microscopy more clearly visualized the intercellular region and the nucleus of the cell compared to the confocal microscopy. The fibrous structures in the dermis were more clearly resolved by the confocal microscopy. Numerous cells in papillary dermal layer, as deep as $100\;{\mu}m$, were observed in both CLSM and two-photon microscopy. While most previous studies focused on fibrous structure imaging (collagen and elastin fiber) in the dermis, we demonstrated that the combined imaging with the CLSM and two-photon microscopy can be applied for the non-invasive study of the population, distribution and metabolism of papillary dermal cells in skin.

Advances in higher-order chromatin architecture: the move towards 4D genome

  • Jung, Namyoung;Kim, Tae-Kyung
    • BMB Reports
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    • v.54 no.5
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    • pp.233-245
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    • 2021
  • In eukaryotes, the genome is hierarchically packed inside the nucleus, which facilitates physical contact between cis-regulatory elements (CREs), such as enhancers and promoters. Accumulating evidence highlights the critical role of higher-order chromatin structure in precise regulation of spatiotemporal gene expression under diverse biological contexts including lineage commitment and cell activation by external stimulus. Genomics and imaging-based technologies, such as Hi-C and DNA fluorescence in situ hybridization (FISH), have revealed the key principles of genome folding, while newly developed tools focus on improvement in resolution, throughput and modality at single-cell and population levels, and challenge the knowledge obtained through conventional approaches. In this review, we discuss recent advances in our understanding of principles of higher-order chromosome conformation and technologies to investigate 4D chromatin interactions.