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.

scholarly journals Microtubules in Candida albicans Hyphae Drive Nuclear Dynamics and Connect Cell Cycle Progression to Morphogenesis

2005 ◽  
Vol 4 (10) ◽  
pp. 1697-1711 ◽  
Author(s):  
Kenneth R. Finley ◽  
Judith Berman
Keyword(s):  
Cell Cycle ◽  
Candida Albicans ◽  
Basal Cell ◽  
Time Lapse ◽  
Nuclear Migration ◽  
Microtubule Dynamics ◽  
Growth Defect ◽  
Cell Cortex ◽  
Long Distance ◽  
Time Lapse Microscopy

ABSTRACT Candida albicans is an opportunistic fungal pathogen whose virulence is related to its ability to switch between yeast, pseudohyphal, and true-hyphal morphologies. To ask how long-distance nuclear migration occurs in C. albicans hyphae, we identified the fundamental properties of nuclear movements and microtubule dynamics using time-lapse microscopy. In hyphae, nuclei migrate to, and divide across, the presumptive site of septation, which forms 10 to 15 μm distal to the basal cell. The mother nucleus returns to the basal cell, while the daughter nucleus reiterates the process. We used time-lapse microscopy to identify the mechanisms by which C. albicans nuclei move over long distances and are coordinated with hyphal morphology. We followed nuclear migration and spindle dynamics, as well as the time and position of septum specification, defined it as the presumptum, and established a chronology of nuclear, spindle, and morphological events. Analysis of microtubule dynamics revealed that premitotic forward nuclear migration is due to the repetitive sliding of astral microtubules along the cell cortex but that postmitotic forward and reverse nuclear migrations are due primarily to spindle elongation. Free microtubules exhibit cell cycle regulation; they are present during interphase and disappear at the time of spindle assembly. Finally, a growth defect in strains expressing Tub2-green fluorescent protein revealed a connection between hyphal elongation and the nuclear cell cycle that is coordinated by hyphal length and/or volume.

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.

Deep learning for tracking of intracellular vesicles in time-lapse microscopy images

2021 ◽  
Author(s):  
Zhichao Liu ◽  
Luhong Jin ◽  
Alexander Nedzved ◽  
Sergey Ablameyko ◽  
Yingke Xu
Keyword(s):  
Deep Learning ◽  
Time Lapse ◽  
Time Lapse Microscopy ◽  
Intracellular Vesicles ◽  
Microscopy Images

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).

Using Deep Learning to Identify Cell and Particle in Live-Cell Time-lapse Images

2018 ◽  
Author(s):  
Hui-Jun Cheng ◽  
Chun-Yuan Lin ◽  
Cheng-Xian Wu ◽  
Che-Lun Hung ◽  
Wei-Hsiang Chen ◽  
...  
Keyword(s):  
Deep Learning ◽  
Time Lapse ◽  
Live Cell
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