scholarly journals Functional Analysis of Human Microtubule-based Motor Proteins, the Kinesins and Dyneins, in Mitosis/Cytokinesis Using RNA Interference

2005 ◽  
Vol 16 (7) ◽  
pp. 3187-3199 ◽  
Author(s):  
Changjun Zhu ◽  
Jian Zhao ◽  
Marina Bibikova ◽  
Joel D. Leverson ◽  
Ella Bossy-Wetzel ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Functional Analysis ◽  
Motor Proteins ◽  
Time Lapse ◽  
Rnase Iii ◽  
Immunofluorescence Analysis ◽  
Cellular Processes ◽  
Time Lapse Microscopy ◽  
Chromosome Congression ◽  
Spindle Dynamics

Microtubule (MT)-based motor proteins, kinesins and dyneins, play important roles in multiple cellular processes including cell division. In this study, we describe the generation and use of an Escherichia coli RNase III-prepared human kinesin/dynein esiRNA library to systematically analyze the functions of all human kinesin/dynein MT motor proteins. Our results indicate that at least 12 kinesins are involved in mitosis and cytokinesis. Eg5 (a member of the kinesin-5 family), Kif2A (a member of the kinesin-13 family), and KifC1 (a member of the kinesin-14 family) are crucial for spindle formation; KifC1, MCAK (a member of the kinesin-13 family), CENP-E (a member of the kinesin-7 family), Kif14 (a member of the kinesin-3 family), Kif18 (a member of the kinesin-8 family), and Kid (a member of the kinesin-10 family) are required for chromosome congression and alignment; Kif4A and Kif4B (members of the kinesin-4 family) have roles in anaphase spindle dynamics; and Kif4A, Kif4B, MKLP1, and MKLP2 (members of the kinesin-6 family) are essential for cytokinesis. Using immunofluorescence analysis, time-lapse microscopy, and rescue experiments, we investigate the roles of these 12 kinesins in detail.

Continuous Measurement of Biological Noise in Escherichia Coli Using Time-lapse Microscopy

10.3791/61290 ◽  
2021 ◽  
Author(s):  
Einel A. Chaimovitz ◽  
Evgeniy Reznik ◽  
Mouna Habib ◽  
Netanel Korin ◽  
Ramez Daniel
Keyword(s):  
Escherichia Coli ◽  
Continuous Measurement ◽  
Time Lapse ◽  
Biological Noise ◽  
Time Lapse Microscopy

scholarly journals The lighthouse at the end of the chromosome*

F1000Research ◽  
2015 ◽  
Vol 4 ◽  
pp. 1427 ◽  
Author(s):  
Yahya Benslimane ◽  
Lea Harrington
Keyword(s):  
Fluorescence Microscopy ◽  
Single Molecule ◽  
Spatial Organization ◽  
Three Dimensional ◽  
Time Lapse ◽  
Living Cells ◽  
Telomere Elongation ◽  
Cellular Processes ◽  
Time Lapse Microscopy

Fluorescence microscopy can be used to assess the dynamic localization and intensity of single entities in vitro or in living cells. It has been applied with aplomb to many different cellular processes and has significantly enlightened our understanding of the heterogeneity and complexity of biological systems. Recently, high-resolution fluorescence microscopy has been brought to bear on telomeres, leading to new insights into telomere spatial organization and accessibility, and into the mechanistic nuances of telomere elongation. We provide a snapshot of some of these recent advances with a focus on mammalian systems, and show how three-dimensional, time-lapse microscopy and single-molecule fluorescence shine a new light on the end of the chromosome.

scholarly journals Microfluidic Cultivation and Laser Tweezers Raman Spectroscopy of E. coli under Antibiotic Stress

2018 ◽  
Author(s):  
Zdeněk Pilát ◽  
Silvie Bernatová ◽  
Jan Ježek ◽  
Johanna Kirchhoff ◽  
Astrid Tannert ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Raman Spectroscopy ◽  
Raman Spectra ◽  
Cell Elongation ◽  
Time Lapse ◽  
Laser Tweezers ◽  
E Coli ◽  
Cellular Physiology ◽  
Time Lapse Microscopy ◽  
Metabolic States

Analyzing the cells in various body fluids can greatly deepen the understanding of the mechanisms governing the cellular physiology. Because of the variability of physiological and metabolic states, it is important to be able to perform such studies on individual cells. Therefore, we developed an optofluidic system in which we precisely manipulated and monitored individual cells of Escherichia coli. We used laser tweezers Raman spectroscopy (LTRS) in a microchamber chip to manipulate and analyze individual E. coli cells. We subjected the cells to antibiotic cefotaxime, and we observed the changes by the time-lapse microscopy and Raman spectroscopy. We found observable changes in the cellular morphology (cell elongation) and in Raman spectra, which were consistent with other recently published observations. We tested the capabilities of the optofluidic system and found it to be a reliable and versatile solution for this class of microbiological experiments.

scholarly journals SpinX: Time-resolved 3D Analysis of Mitotic Spindle Dynamics using Deep Learning Techniques and Mathematical Modelling

2021 ◽  
Author(s):  
David Dang ◽  
Christoforos Efstathiou ◽  
Dijue Sun ◽  
Nishanth Sastry ◽  
Viji M Draviam
Keyword(s):  
Deep Learning ◽  
Mathematical Object ◽  
Time Lapse ◽  
Cell Cortex ◽  
3D Analysis ◽  
Time Resolved ◽  
3 Dimensional ◽  
Time Lapse Microscopy ◽  
Exciting Opportunity ◽  
Spindle Dynamics

Time-lapse microscopy movies have transformed the study of subcellular dynamics. However, manual analysis of movies can introduce bias and variability, obscuring important insights. While automation can overcome such limitations, spatial and temporal discontinuities in time-lapse movies render methods such as object segmentation and tracking difficult. Here we present SpinX, a framework for reconstructing gaps between successive frames by combining Deep Learning and mathematical object modelling. By incorporating expert feedback through selective annotations, SpinX identifies subcellular structures, despite confounding neighbour-cell information, non-uniform illumination and variable marker intensities. The automation and continuity introduced allows precise 3-Dimensional tracking and analysis of spindle movements with respect to the cell cortex for the first time. We demonstrate the utility of SpinX using distinct spindle markers and drug treatments. In summary, SpinX provides an exciting opportunity to study spindle dynamics in a sophisticated way, creating a framework for step changes in studies using time-lapse microscopy.

Experience of using time-lapse microscopy in IVF and ICSI programs

2020 ◽  
pp. 47-50
Author(s):  
N. V. Saraeva ◽  
N. V. Spiridonova ◽  
M. T. Tugushev ◽  
O. V. Shurygina ◽  
A. I. Sinitsyna
Keyword(s):  
Pregnancy Rate ◽  
Embryo Transfer ◽  
Selection Procedure ◽  
Time Lapse ◽  
Single Embryo Transfer ◽  
Main Group ◽  
Clinical Pregnancy ◽  
Control Group ◽  
Time Lapse Microscopy

In order to increase the pregnancy rate in the assisted reproductive technology, the selection of one embryo with the highest implantation potential it is very important. Time-lapse microscopy (TLM) is a tool for selecting quality embryos for transfer. This study aimed to assess the benefits of single-embryo transfer of autologous oocytes performed on day 5 of embryo incubation in a TLM-equipped system in IVF and ICSI programs. Single-embryo transfer following incubation in a TLM-equipped incubator was performed in 282 patients, who formed the main group; the control group consisted of 461 patients undergoing single-embryo transfer following a traditional culture and embryo selection procedure. We assessed the quality of transferred embryos, the rates of clinical pregnancy and delivery. The groups did not differ in the ratio of IVF and ICSI cycles, average age, and infertility factor. The proportion of excellent quality embryos for transfer was 77.0% in the main group and 65.1% in the control group (p = 0.001). In the subgroup with receiving eight and less oocytes we noted the tendency of receiving more quality embryos in the main group (р = 0.052). In the subgroup of nine and more oocytes the quality of the transferred embryos did not differ between two groups. The clinical pregnancy rate was 60.2% in the main group and 52.9% in the control group (p = 0.057). The delivery rate was 45.0% in the main group and 39.9% in the control group (p > 0.050).

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