• Title/Summary/Keyword: Fluorescent microscopy

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Simple Fabrication of Green Emission and Water-Resistant CsPbBr3 Encapsulation Using Commercial Glass Frits (상업용 유리프릿의 소결 공정을 이용한 내수성을 갖는 CsPbBr3/Glass 세라믹 복합체의 제작)

  • Mun, Na-eun;Kim, Sunghoon
    • Korean Journal of Materials Research
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    • v.31 no.1
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    • pp.54-59
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    • 2021
  • In this work, narrow-band green-emitting CsPbBr3 particles are embedded in commercialized glass composites by a facile dry process. By optimizing the method through sintering in glass frit (GF) composites including CsBr and PbBr2, used as precursors, the encapsulation of CsPbBr3 particles made them waterproof with green fluorescence. To improve the fluorescent properties by reducing aggregation of CsPbBr3, fumed silica (FS) is additionally used to help particles avoid bulking up in the glass matrix. The CsPbBr3 perovskite/glass composites are characterized using scanning electron microscopy (SEM) images and energy-dispersive X-ray spectroscopy (EDS) maps, which support the existence of CsPbBr3 particles in the glass matrix. The photoluminescence (PL) properties demonstrate that the emission spectrum peak, full width at half maximum (FWHM), and photoluminescence quantum yield (PLQY) values are 519 nm, 17 nm, and 17.7 %. We also confirm the water-resistant properties. To enhance water/moisture stability, the composite sample is put directly into water, with its PLQY monitored periodically under UV light.

Antioxidant and Antibacterial Activity of Caprylic Acid Vanillyl Ester Produced by Lipase-Mediated Transesterification

  • Kim, Jin Ju;Kim, Hyung Kwoun
    • Journal of Microbiology and Biotechnology
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    • v.31 no.2
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    • pp.317-326
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    • 2021
  • Vanillyl alcohol (VA), which is abundant in Vanilla bean, has strong antioxidant activity. However, the use of VA in the food and cosmetics industries is limited, due to its low solubility in emulsion or organic solvents. Meanwhile, medium chain fatty acids and medium chain monoglycerides have antibacterial activity. We synthesized butyric acid vanillyl ester (BAVE) or caprylic acid vanillyl ester (CAVE) from VA with tributyrin or tricaprylin through transesterification reaction using immobilized lipases. BAVE and CAVE scavenged 2,2-diphenyl-1-picrylhydrazyl radicals in organic solvents. In addition, BAVE and CAVE decreased the production rate of conjugated diene and triene in the menhaden oil-in-water emulsion system. While BAVE showed no antibacterial activity, CAVE showed antibacterial activity against food spoilage bacteria, including Bacillus coagulans. In this study, the antibacterial activity of vanillyl ester with medium chain fatty acid was first revealed. Zeta potential measurements confirmed that BAVE and CAVE were inserted into B. coagulans membrane. In addition, the propidium iodide uptake assay and fluorescent microscopy showed that CAVE increased B. coagulans membrane permeability. Therefore, CAVE is expected to play an important role in the food and cosmetics industries as a bi-functional material with both antioxidant and antibacterial activities.

Sclerotiorin: a Novel Azaphilone with Demonstrated Membrane Targeting and DNA Binding Activity against Methicillin-Resistant Staphylococcus aureus

  • Dasagrandhi, Chakradhar;Pandith, Anup;Imran, Khalid
    • Microbiology and Biotechnology Letters
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    • v.48 no.4
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    • pp.429-438
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    • 2020
  • The emergence of multi-drug resistant, pathogenic methicillin-resistant Staphylococcus aureus (MRSA) is a threat to global health and has created a need for novel functional therapeutic agents. In this study, we evaluated the underlying mechanisms of the anti-MRSA effect of an azaphilone pigment, sclerotiorin (SCL) from Penicillium sclerotiorum. The antimicrobial activity of SCL was evaluated using agar disc diffusion, broth microdilution, time-kill assays and biophysical studies. SCL exhibits selective activity against Gram positive bacteria including MRSA (range, MIC = 128-1028 ㎍/ml) and exhibited rapid bactericidal action against MRSA with a > 4 log reduction in colony forming units within three hours of administration. Biophysical studies, using fluorescent probes and laser or electron microscopy, demonstrated a SCL dose-dependent alternation in membrane potential (62.6 ± 5.0.4% inhibition) and integrity (> 95 ± 2.3%), and the release of UV260 absorbing materials within 60 min (up to 3.2 fold increase, p < 0.01) of exposure. Further, SCL localized to the cytoplasm and hydrolyzed plasmid DNA. While in vitro checkerboard studies revealed that SCL potentiated the antimicrobial activity of topical antimicrobials such as polymixin, neomycin, and bacitracin (Fractional Inhibitory Concentration Index range, 0.26-0.37). Taken together these results suggest that SCL targets the membrane and DNA of MRSA to facilitate its anti-MRSA antimicrobial effect.

Characterizing Organelles in Live Stem Cells Using Label-Free Optical Diffraction Tomography

  • Kim, Youngkyu;Kim, Tae-Keun;Shin, Yeonhee;Tak, Eunyoung;Song, Gi-Won;Oh, Yeon-Mok;Kim, Jun Ki;Pack, Chan-Gi
    • Molecules and Cells
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    • v.44 no.11
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    • pp.851-860
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    • 2021
  • Label-free optical diffraction tomography (ODT), an imaging technology that does not require fluorescent labeling or other pre-processing, can overcome the limitations of conventional cell imaging technologies, such as fluorescence and electron microscopy. In this study, we used ODT to characterize the cellular organelles of three different stem cells-namely, human liver derived stem cell, human umbilical cord matrix derived mesenchymal stem cell, and human induced pluripotent stem cell-based on their refractive index and volume of organelles. The physical property of each stem cell was compared with that of fibroblast. Based on our findings, the characteristic physical properties of specific stem cells can be quantitatively distinguished based on their refractive index and volume of cellular organelles. Altogether, the method employed herein could aid in the distinction of living stem cells from normal cells without the use of fluorescence or specific biomarkers.

Apoptin gene delivery by a PAMAM dendrimer modified with a nuclear localization signal peptide as a gene carrier for brain cancer therapy

  • Bae, Yoonhee;Lee, Jell;Kho, Changwon;Choi, Joon Sig;Han, Jin
    • The Korean Journal of Physiology and Pharmacology
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    • v.25 no.5
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    • pp.467-478
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    • 2021
  • In this study, we aimed to synthesize PAMAMG3 derivatives (PAMAMG3-KRRR and PAMAMG3-HKRRR), using KRRR peptides as a nuclear localization signal and introduced histidine residues into the KRRR-grafted PAMAMG3 for delivering a therapeutic, carcinoma cell-selective apoptosis gene, apoptin into human primary glioma (GBL-14) cells and human dermal fibroblasts. We examined their cytotoxicity and gene expression using luciferase activity and enhanced green fluorescent protein PAMAMG3 derivatives in both cell lines. We treated cells with PAMAMG3 derivative/apoptin complexes and investigated their intracellular distribution using confocal microscopy. The PAMAMG3-KRRR and PAMAMG3-HKRRR dendrimers were found to escape from endolysosomes into the cytosol. The JC-1 assay, glutathione levels, and Annexin V staining results showed that apoptin triggered cell death in GBL-14 cells. Overall, these findings indicated that the PAMAMG3-HKRRR/apoptin complex is a potential candidate for an effective nonviral gene delivery system for brain tumor therapy in vitro.

Lipofuscin Granule Accumulation Requires Autophagy Activation

  • Seon Beom Song;Woosung Shim;Eun Seong Hwang
    • Molecules and Cells
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    • v.46 no.8
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    • pp.486-495
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    • 2023
  • Lipofuscins are oxidized lipid and protein complexes that accumulate during cellular senescence and tissue aging, regarded as markers for cellular oxidative damage, tissue aging, and certain aging-associated diseases. Therefore, understanding their cellular biological properties is crucial for effective treatment development. Through traditional microscopy, lipofuscins are readily observed as fluorescent granules thought to accumulate in lysosomes. However, lipofuscin granule formation and accumulation in senescent cells are poorly understood. Thus, this study examined lipofuscin accumulation in human fibroblasts exposed to various stressors. Our results substantiate that in glucose-starved or replicative senescence cells, where elevated oxidative stress levels activate autophagy, lipofuscins predominately appear as granules that co-localize with autolysosomes due to lysosomal acidity or impairment. Meanwhile, autophagosome formation is attenuated in cells experiencing oxidative stress induced by a doxorubicin pulse and chase, and lipofuscin fluorescence granules seldom manifest in the cytoplasm. As Torin-1 treatment activates autophagy, granular lipofuscins intensify and dominate, indicating that autophagy activation triggers their accumulation. Our results suggest that high oxidative stress activates autophagy but fails in lipofuscin removal, leaving an abundance of lipofuscin-filled impaired autolysosomes, referred to as residual bodies. Therefore, future endeavors in treating lipofuscin pathology-associated diseases and dysfunctions through autophagy activation demand meticulous consideration.

Applications of the Scanning Electron Microscope (주사형(走査型) 전자현미경(電子顯微鏡)의 응용분야(應用分野))

  • Kim, Yong-Nak
    • Applied Microscopy
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    • v.2 no.1
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    • pp.39-46
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    • 1972
  • There are many kinds of microscopes suitable for general studies; optical microscopes(OM), conventional transmission electron microscopes (TEM), and scanning electron microscopes(SEM). The optical microscopes and the conventional transmission electron microscopes are very familiar. The images of these microscopes are directly formed on an image plane with one or more image forming lenses. On the other hand, the image of the scanning electron microscope is formed on a fluorescent screen of a cathode ray tube using a scanning system similar to television technique. In this paper, the features and some applications of the scanning electron microscope will be discussed briefly. The recently available scanning electron microscope, combining a resolution of about $200{\AA}$ with great depth of field, is favorable when compared to the replica technique. It avoids the problem of specimen damage and the introduction of artifacts. In addition, it permits the examination of many samples that can not be replicated, and provides a broader range of information. The scanning electron microscope has found application in diverse fields of study including biology, chemistry, materials science, semiconductor technology, and many others. In scanning electron microscopy, the secondary electron method. the backscattererd electron method, and the electromotive force method are most widely used, and the transmitted electron method will become more useful. Change-over of magnification can be easily done by controlling the scanning width of the electron probe. It is possible. to continuously vary the magnification over the range from 100 times to 1.00,000 times without readjustment of focusing. Conclusion: With the development of a scanning. electron microscope, it is now possible to observe almost all-information produced through interactions between substances and electrons in the form of image. When the probe is properly focused on the specimen, changing magnification of specimen orientation does not require any change in focus. This is quite different from the conventional transmission electron microscope. It is worthwhile to note that the typical probe currents of $10^{-10}$ to $10^{-12}\;{\AA}$ are for below the $10^{-5}$ to $10^{-7}\;{\AA}$ of a conventional. transmission microscope. This reduces specimen contamination and specimen damage due to heatings. Outstanding features of the scanning electron microscope include the 'stereoscopic observation of a bulky or fiber specimen in high resolution' and 'observation of potential distribution and electromotive force in semiconductor devices'.

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Site-Specific Recombination by the Integrase MJ1 on Mammalian Cell (동물 세포 내에서 MJ1 인티그라제에 의한 부위 특이적 재조합)

  • Kim, Hye-Young;Yoon, Bo-Hyun;Chang, Hyo-Ihl
    • Microbiology and Biotechnology Letters
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    • v.39 no.4
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    • pp.337-344
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    • 2011
  • Integrase MJ1 from the bacteriophage ${\Phi}FC1$ carries out recombination between two DNA sequences (the phage attachment site, attP and the bacterial attachment site, attB) in NIH3T3 mouse cells. In this study, the integration vector containing attP, attB and the integrase gene MJ, was constructed. The integration mediated by integrase MJ1 in Escherichia coli led to excision of LacZ. Therefore, the frequency of integration was measured by the counting of the white colony, which is detectable on X-Gal plates. The extrachromosomal integration in NIH3T3 mouse cells was monitored by the expression of the green fluorescent protein (GFP) as a reporter. To demonstrate integration mediated integrase MJ1 in NIH3T3 cells, vectors containing attP and attB were co-transfected into NIH3T3 cells. The integration was confirmed by fluorescent microscopy. The expression of GFP was induced in NIH3T3 cells expressing MJ1 without accessory factors. By contrast, the excision mediated by the MJ1 between attR and attL had no effect on the expression of GFP. These results suggest that integrase MJ1 may enable a variety of genomic modifications for research and therapeutic purposes in higher living cells.

Osseointegration of Implants in Rabbit Bone with a Low Calcium Diet and Irradition (저칼슘식이 투여와 방사선조사가 임프란트의 골유착에 미치는 영향에 관한 실험적 연구)

  • Kim So-Jung;Hwang Eui-Hwan;Lee Sang-Rae
    • Imaging Science in Dentistry
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    • v.30 no.1
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    • pp.33-48
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    • 2000
  • Purpose : To investigate osseointergration of titanium implants into the tibia of rabbits, which were fed a low calcium diet and irradiated. Materials and Methods : To prepare the experimental model, control group was fed a normal diet and experimental group was fed a low calcium diet for 4 weeks. And then, titanium implants were inserted into the tibia of each rabbit. Experimental group was subdivided into two groups; low calcium diet/non-irradiation group and low calcium diet/irradiation group. The low calcium diet/irradiation group was irradiated with a single absorbed dose of 15 Gy at the 5th postoperative day. At 12, 19, 33, 47, and 61 days after implantation (1, 2, 4, 6, and 8 weeks after irradiation), the bone formation in the bone-implant interface area was examined by light microscopy and fluorescent microscopy. Results and Conclusions: 1. In the control group, there began to form woven bone in the bone-implant interface area at 12 days after implantation. As the experimental time was going on, the amount of bone which was in contact with the implant was increased. 2. In the low calcium diet/non-irradiation group, there began to form woven bone in the bone-implant interface area at 19 days after implantation. Although the amount of bone which was in contact with the implant was increased as the experimental time was going on, the extent of increased bone was weak as compared with control group. 3. In the low calcium diet/irradiation group, there began to form woven bone incompletely in the bone-implant interface area at 19 days after implantation, but there were vascular connective tissues in the bone-implant interface area over the entire experimental period. 4. In the control group and low calcium diet/non-irradiation group, bone labeling bands were observed at 33 days after implantation, which suggests that the bone formation and remodeling was in process, but interstitial bone remodeling was not observed in the low calcium diet/irradiation group.

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Healing of the Bone around Hydroxyapatite-Coated Implants without Primary Bone Contact (초기 골 접촉이 없는 수산화 인회석 피복 임프란트 주위 골의 치유)

  • Cho, Hyung-Soo;Shin, Kwang-Yong;Kim, Heung-Joong;Park, Joo-Cheol;Han, Kyung-Yoon;Kim, Byung-Ock
    • Journal of Periodontal and Implant Science
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    • v.29 no.2
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    • pp.415-433
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    • 1999
  • Implant stability is the key to long-term successful outcome for osseointegrated implants. To evaluate the initial healing response of bone around HA-coated implants without primary bone contact. 21 HA-coated thread type implants(STERI-OSS?) were placed in the femurs of 5 mongrel dogs, about 1-year old. Implants, 8 mm in length and 3.8mm(experimental 1group), 5.0mm(experimental 2group) and 6.0mm(control group) in diameter, were inserted after 3 holes of 6.0mm in diameter and 10mm in depth were prepared in the surgical sites each dog. Implants were supported by only nonresorbable membrane($Teflon^{(R)}$), in order to prevent the ingrowth of upper soft tissue into the gap between bone and implant, and to maintain each implant to be positioned in the center of the drilled hole. 9 implants with different diameters were inserted in 3 dogs for histologic observation, and 12 implants were inserted in 2 dogs for mobility test and removal torque test. Fluorescent dyes were injected for the observation of new bone formation in order of $Terramycin^{(R)}$, Arizarin $Red^{(R)}$, and $Calcein^{(R)}$ at an interval of 2 weeks. 3 dogs were sacrificed for histologic observation at 4, 8, and 12-week after placement. Light microscopy and confocal laser scanning microscopy were used to qualitatively characterize the bone around HA-coated implant. 2 dogs were sacrificed for mobility test($Periotest^{(R)}$, Simens AG, Bensheim, Germany) and removal torque test($Autograph^{(R)}$ AGS-1000D series, Japan) at 8 and 12-week after placement The results were as follows: 1. Histologic observation showed that osseointegration occurred to both control and experimental groups as time lapse, but delayed bone healing was revealed in 3.8mm group (experimental 1group), compared to contrtol group and 5.0mm group (experimental 2group). 2. The mobility test showed that the experimental groups had no distinguishable movement during experimental periods of 8 and 12-week, and there was no difference in mobility depending on the gap between bone and implant, and time lapse. 3. The removal torque forces were increased depended on the gaps decreasing between bone and implant, and time lapse. The results suggest that HA-coated implant without primary bone contact, based on guided bone regeneration could obtain its stability in all experimental groups as time lapse, but bone healing was delayed in experimental group of 3.8mm. And the results suggested that studies on correlationship between mobility test and removal torque test for implant stability would be necessary.

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