• 제목/요약/키워드: live cell

검색결과 392건 처리시간 0.033초

Anabaena flos-aquae 에서의 세포사멸계수(Cell Death Constant)의 측정 (Measurement of Cell Death Constant in Anabaena flos-aquae (Cyanophyceae) by the Molecular Probe)

  • 오인혜
    • The Korean Journal of Ecology
    • /
    • 제20권3호
    • /
    • pp.169-173
    • /
    • 1997
  • The measurement of cell death constant in Anabaena flos-aquae was tested by the Live/Dead BacLight Viability kit(Molecular Probes Co., Seatle, WA). When the Live/Dead BacLight Viability kit was applied to Anabaena flos-aquae, the cells with intact cell membranes(live cells) stained fluorescent green, while the cell with damaged membranes(dead cells) stained fluorescent red and the background remained virtually nonfluorescent. The rations of live : dead cells in the cell suspension were controlled artifically and Live/Dead BacLight Viability kit was applied to them. The ratios of green:red fluorescent cells in the cell suspension were the same as those of live : dead cells controlled artifically. It was also approved by the fluorescence emission. The cell death constant was measured in the P-limited Anabaena flos-aquae chemostal culture in the N-fixing and $KNO_3-supplied$ conditions. The culture in N-fixing chemostat had a dead cell proportion of 1.2% at the growth rate of 0.7/day and increased to 2.6% at the growth rate of 0.3/day. The cell death constant of N-fixing culture was 0.008/day.There was a same trend in the $KNO_3-supplied$ chemostat culture. The proportion of dead cell was 1.6% of dead cell proportion at the growth rate of 0.7/day and increased to 4.3% at the growth rate of 0.3/day.

  • PDF

생혈액검사상 적혈구의 연전현상과 피로도와의 상관관계 (Correlation between Rouleau Condition of Red Blood Cell on Live Blood Analysis and Degree of Fatigue)

  • 신승우;이종수
    • 대한추나의학회지
    • /
    • 제2권1호
    • /
    • pp.159-168
    • /
    • 2001
  • Objectives : It is known that rouleau condition of red blood cell on live blood analysis is related to degree of fatigue. This study was planed to prove the correlation between rouleau condition of red blood cell and degree of fatigue by using the questionnaire 'Symptom Table on Fatigue Perception' that had been verified before. Methods : We analyzed the correlation between rouleau condition and degree of fatigue by calculating ratio of rouleau of red blood cell after measuring degree of fatigue by the questionnaire to the people who had not evidence of illness on health examination and did not take any medicine. Results and Conclusions : This study showed a significance of positive correlation(0.464) between physical fatigue and rouleau condition of red blood cell.

  • PDF

Imaging Single-mRNA Localization and Translation in Live Neurons

  • Lee, Byung Hun;Bae, Seong-Woo;Shim, Jaeyoun Jay;Park, Sung Young;Park, Hye Yoon
    • Molecules and Cells
    • /
    • 제39권12호
    • /
    • pp.841-846
    • /
    • 2016
  • Local protein synthesis mediates precise spatio-temporal regulation of gene expression for neuronal functions such as long-term plasticity, axon guidance and regeneration. To reveal the underlying mechanisms of local translation, it is crucial to understand mRNA transport, localization and translation in live neurons. Among various techniques for mRNA analysis, fluorescence microscopy has been widely used as the most direct method to study localization of mRNA. Live-cell imaging of single RNA molecules is particularly advantageous to dissect the highly heterogeneous and dynamic nature of messenger ribonucleoprotein (mRNP) complexes in neurons. Here, we review recent advances in the study of mRNA localization and translation in live neurons using novel techniques for single-RNA imaging.

락토바실러스 아시도필러스 CBT LA1 생균과 사균체의 세포부착력, 자가응집력, 소수성 상호작용력, LPS 결합력에 대한 평가 (Assessment of cell adhesion, cell surface hydrophobicity, autoaggregation, and lipopolysaccharide-binding properties of live and heat-killed Lactobacillus acidophilus CBT LA1)

  • 신주현;이중수;서재구
    • 미생물학회지
    • /
    • 제51권3호
    • /
    • pp.241-248
    • /
    • 2015
  • 프로바이오틱스에 대한 연구는 주로 생균의 효과가 많이 알려져 있지만 가열 처리된 유산균인 사균체의 기능에 대한 연구도 활발히 이루어지고 있다. 본 연구에서는 락토바실러스 아시도필러스 CBT LA1의 사균체의 장관장벽 기능에 대하여 in vitro, in vivo에서 실험하였다. 이를 위하여, 세포표면 소수성 상호작용력(cell surface hydrophobicity), 자가응집력(autoaggregation), 세포에 부착하는 능력(cell adhesion)과 자가응집력(autoaggregation), LPS와의 결합력을 조사하였다. 또한 HT-29 장상피세포에서 LPS로 유도되는 IL-8의 발현을 억제하는 효과를 조사하였다. CBT LA1을 80도에서 121도까지 10분 동안 열을 처리하였을 때, 80도에서 열을 처리한 CBT LA1 사균체가 가장 높은 세포에 부착하는 능력을 보여 주었다. CBT LA1 생균과 비교했을 때, 80도에서 열을 처리한 CBT LA1 사균체는 높은 LPS와의 결합력, 소수성 상호작용력, 자가응집력, HT-29 세포에 부착하는 능력과 IL-8의 발현을 억제하는 능력을 보여주었다. In vivo 실험에서 FITC로 label된 LPS를 투여하였을 때 16시간 후, CBT LA1 사균체를 섭취한 동물의 장관 내 LPS가 가장 많이 제거되었다. 이러한 연구 결과들은 CBT LA1 생균처럼 CBT LA1 사균체도 장관장벽 기능을 가지며 이는, 파마바이오틱스로서 그 가능성을 시사한다.

분자 비컨을 이용한 살아 있는 세포에서 단일클론항체 경쇄와 중쇄 mRNA 검출에 의한 세포주 선별방법 (Live Cell Detection of Monoclonal Antibody Light and Heavy Chain mRNAs using Molecular Beacons)

  • 정승아;이원종
    • KSBB Journal
    • /
    • 제31권1호
    • /
    • pp.33-39
    • /
    • 2016
  • Developing the method for the selection of animal cell line producing therapeutic monoclonal antibody (mAb) is invaluable as its market is rapidly growing. Although the quality of produced mAb is as important as quantity, however there is no method developed for the selective screening of cell lines on the basis of both quantity and quality. From recent reports, the ratio of light and heavy chain mRNAs of mAb in the cell is a key parameter for the indication of product quality. Therefore, it is obvious that developing the novel method that can detect both light and heavy chain mRNAs in single live cell will provide unprecedented opportunities in bio-industry. Here, we have constructed oligonucleotide probes, molecular beacons for the detection of light or heavy chain mRNAs, respectively, in the live cells producing mAbs. Both beacons showed increased fluorescent intensity after transient transfection of plasmid expressing mAbs analyzed by fluorometer. Flow cytometric analysis clearly demonstrated that both molecular beacons can simultaneously detect the expression of light and heavy chain mRNAs of mAb in the same cell. The technique described in the thesis provides the new direction and concept for developing the method for the smart selection of cell lines producing recombinant proteins including therapeutic mAbs.

Time-Lapse Live-Cell Imaging Reveals Dual Function of Oseg4, Drosophila WDR35, in Ciliary Protein Trafficking

  • Lee, Nayoung;Park, Jina;Bae, Yong Chul;Lee, Jung Ho;Kim, Chul Hoon;Moon, Seok Jun
    • Molecules and Cells
    • /
    • 제41권7호
    • /
    • pp.676-683
    • /
    • 2018
  • Cilia are highly specialized antennae-like organelles that extend from the cell surface and act as cell signaling hubs. Intraflagellar transport (IFT) is a specialized form of intracellular protein trafficking that is required for the assembly and maintenance of cilia. Because cilia are so important, mutations in several IFT components lead to human disease. Thus, clarifying the molecular functions of the IFT proteins is a high priority in cilia biology. Live imaging in various species and cellular preparations has proven to be an important technique in both the discovery of IFT and the mechanisms by which it functions. Live imaging of Drosophila cilia, however, has not yet been reported. Here, we have visualized the movement of IFT in Drosophila cilia using time-lapse live imaging for the first time. We found that NOMPB-GFP (IFT88) moves according to distinct parameters depending on the ciliary segment. NOMPB-GFP moves at a similar speed in proximal and distal cilia toward the tip (${\sim}0.45{\mu}m/s$). As it returns to the ciliary base, however, NOMPB-GFP moves at ${\sim}0.12{\mu}m/s$ in distal cilia, accelerating to ${\sim}0.70{\mu}m/s$ in proximal cilia. Furthermore, while live imaging NOMPB-GFP, we observed one of the IFT proteins required for retrograde movement, Oseg4 (WDR35), is also required for anterograde movement in distal cilia. We anticipate our time-lapse live imaging analysis technique in Drosophila cilia will be a good starting point for a more sophisticated analysis of IFT and its molecular mechanisms.

Use of DNA-Specific Anthraquinone Dyes to Directly Reveal Cytoplasmic and Nuclear Boundaries in Live and Fixed Cells

  • Edward, Roy
    • Molecules and Cells
    • /
    • 제27권4호
    • /
    • pp.391-396
    • /
    • 2009
  • Image-based, high-content screening assays demand solutions for image segmentation and cellular compartment encoding to track critical events - for example those reported by GFP fusions within mitosis, signalling pathways and protein translocations. To meet this need, a series of nuclear/cytoplasmic discriminating probes have been developed: DRAQ5$^{TM}$ and CyTRAK Orange$^{TM}$. These are spectrally compatible with GFP reporters offering new solutions in imaging and cytometry. At their most fundamental they provide a convenient fluorescent emission signature which is spectrally separated from the commonly used reporter proteins (e.g. eGFP, YFP, mRFP) and fluorescent tags such as Alexafluor 488, fluorescein and Cy2. Additionally, they do not excite in the UV and thus avoid the complications of compound UV-autofluorescence in drug discovery whilst limiting the impact of background sample autofluorescence. They provide a convenient means of stoichiometrically labelling cell nuclei in live cells without the aid of DMSO and can equally be used for fixed cells. Further developments have permitted the simultaneous and differential labelling of both nuclear and cytoplasmic compartments in live and fixed cells to clearly render the precise location of cell boundaries which may be beneficial for quantitative expression measurements, cell-cell interactions and most recently compound in vitro toxicology testing.

Visualization of Gene Transfer into Live Cells Using Fluorescent Semiconductor Nanocrystals

  • 김중경;임선희;이용구;신영식;정찬일;장준근;유정열
    • 한국가시화정보학회:학술대회논문집
    • /
    • 한국가시화정보학회 2003년도 추계학술대회 논문집
    • /
    • pp.81-82
    • /
    • 2003
  • We have developed the method for the conjugation of biotinylated DNA to streptavidin-coated QDs. QD-DNA conjugates and a high-sensitive fluorescence imaging technique are adopted to visualize gene transport across the membrane of the live cell in real time. Endocytotic cellular uptake of oligonucleotide and electrically-mediated plasmid DNA transfer into the live cell are monitored by a quantitative microscopic imaging system. Long-term kinetic study enables us to reveal the unknown mechanisms and rate-limiting steps of extracellular and intracellular transport of biomolecules. We designed experimental protocols to conjugate the oligonucleotide or the plasmid DNA to commercially available streptavidin-coated QDs. Gel electrophoresis is used to verify the effect of incubation time and the molar ratio of QDs and DNA on the conjugation efficiency. It is possible to fractionate the QD-DNA conjugates according to the DNA concentration and obtain the purified conjugates by a gel extraction technique.

  • PDF