scholarly journals Time-Lapse Imaging of Membrane Traffic in Living Cells

2011 ◽  
Vol 2011 (11) ◽  
pp. pdb.prot066555-pdb.prot066555 ◽  
Author(s):  
E. L. Snapp ◽  
P. Lajoie
Keyword(s):  
Membrane Traffic ◽  
Time Lapse ◽  
Living Cells ◽  
Time Lapse Imaging

scholarly journals Cytoplasmic dynamics of myosin IIA and IIB: spatial ‘sorting’ of isoforms in locomoting cells

1998 ◽  
Vol 111 (15) ◽  
pp. 2085-2095 ◽  
Author(s):  
J. Kolega
Keyword(s):  
Cultured Cells ◽  
Myosin Ii ◽  
Cell Movement ◽  
Leading Edge ◽  
Time Lapse ◽  
Living Cells ◽  
Immunofluorescence Staining ◽  
Spatial Sorting ◽  
Time Lapse Imaging

Different isoforms of non-muscle myosin II have different distributions in vivo, even within individual cells. In order to understand how these different distributions arise, the distribution and dynamics of non-muscle myosins IIA and myosin IIB were examined in cultured cells using immunofluorescence staining and time-lapse imaging of fluorescent analogs. Cultured bovine aortic endothelia contained both myosins IIA and IIB. Both isoforms distributed along stress fibers, in linear or punctate aggregates within lamellipodia, and diffusely around the nucleus. However, the A isoform was preferentially located toward the leading edge of migrating cells when compared with myosin IIB by double immunofluorescence staining. Conversely, the B isoform was enriched in structures at the cells' trailing edges. When fluorescent analogs of the two isoforms were co-injected into living cells, the injected myosins distributed with the same disparate localizations as endogenous myosins IIA and IIB. This indicated that the ability of the myosins to ‘sort’ within the cytoplasm is intrinsic to the proteins themselves, and not a result of localized synthesis or degradation. Furthermore, time-lapse imaging of injected analogs in living cells revealed differences in the rates at which the two isoforms rearranged during cell movement. The A isoform appeared in newly formed structures more rapidly than the B isoform, and was also lost more rapidly when structures disassembled. These observations suggest that the different localizations of myosins IIA and IIB reflect different rates at which the isoforms transit through assembly, movement and disassembly within the cell. The relative proportions of different myosin II isoforms within a particular cell type may determine the lifetimes of various myosin II-based structures in that cell.

scholarly journals Spatial distribution and functional significance of activated vinculin in living cells

2005 ◽  
Vol 169 (3) ◽  
pp. 459-470 ◽  
Author(s):  
Hui Chen ◽  
Daniel M. Cohen ◽  
Dilshad M. Choudhury ◽  
Noriyuki Kioka ◽  
Susan W. Craig
Keyword(s):  
Conformational Change ◽  
Direct Evidence ◽  
Resonance Energy Transfer ◽  
Focal Adhesions ◽  
Resonance Energy ◽  
Time Lapse ◽  
Living Cells ◽  
Actin Binding ◽  
Binding Conformation ◽  
Time Lapse Imaging

Conformational change is believed to be important to vinculin's function at sites of cell adhesion. However, nothing is known about vinculin's conformation in living cells. Using a Forster resonance energy transfer probe that reports on changes in vinculin's conformation, we find that vinculin is in the actin-binding conformation in a peripheral band of adhesive puncta in spreading cells. However, in fully spread cells with established polarity, vinculin's conformation is variable at focal adhesions. Time-lapse imaging reveals a gradient of conformational change that precedes loss of vinculin from focal adhesions in retracting regions. At stable or protruding regions, recruitment of vinculin is not necessarily coupled to the actin-binding conformation. However, a different measure of vinculin conformation, the recruitment of vinexin β by activated vinculin, shows that autoinhibition of endogenous vinculin is relaxed at focal adhesions. Beyond providing direct evidence that vinculin is activated at focal adhesions, this study shows that the specific functional conformation correlates with regional cellular dynamics.

Temporal and Spatial Dynamics of the Actin-Based Cytoskeleton

1990 ◽  
Vol 48 (2) ◽  
pp. 546-546
Author(s):  
D. Lansing Taylor
Keyword(s):  
Spatial Dynamics ◽  
Time Lapse ◽  
Living Cells ◽  
Biomedical Sciences ◽  
3T3 Cells ◽  
Cellular Functions ◽  
Contrast Microscopy ◽  
Interference Contrast Microscopy ◽  
Temporal And Spatial ◽  
Time Lapse Imaging

There has been a renaissance and revolution in the use of light microscopy in the biomedical sciences. The renaissance has been due to the importance of studying the temporal and spatial dynamics of ions, metabolites and macromolecules in living cells and tissues. The revolution has been due to the integration of developments in molecular biology, fluorescent probe chemistry, machine vision, and imaging technology. It is now possible to use the living cell as a microcuvette and to explore the chemical and molecular dynamics responsible for cellular functions.We have been investigating the formation, transport and contraction of stress fibers in Swiss 3T3 cells. Fluorescent analogs of actin, myosin, vinculin and profilin have been investigated in serum deprived cells before, during and after stimulation with thrombin. The activities of these components of the actin-based cytoskeleton have been quantified using time-lapse imaging, fluorescence redistribution after photobleaching, video-enhanced contrast and reflection interference contrast microscopy.

scholarly journals Time-lapse imaging of microRNA activity reveals the kinetics of microRNA activation in single living cells

2017 ◽  
Vol 7 (1) ◽  
Author(s):  
Hideaki Ando ◽  
Matsumi Hirose ◽  
Gen Kurosawa ◽  
Soren Impey ◽  
Katsuhiko Mikoshiba
Keyword(s):  
Time Lapse ◽  
Living Cells ◽  
Single Living Cells ◽  
Time Lapse Imaging ◽  
Kinetics Of ◽  
Single Living

Cleavage of Human Embryos: Options and Diversity

Acta Naturae ◽  
2016 ◽  
Vol 8 (3) ◽  
pp. 88-96
Author(s):  
Yu. K. Doronin ◽  
I. V. Senechkin ◽  
L. V. Hilkevich ◽  
M. A. Kurcer
Keyword(s):  
Time Lapse ◽  
Reproductive Technologies ◽  
Human Embryos ◽  
Imaging Data ◽  
Noninvasive Methods ◽  
Topographic Features ◽  
Time Parameters ◽  
Embryo Cleavage ◽  
Time Lapse Imaging ◽  
Developmental Dynamics

In order to estimate the diversity of embryo cleavage relatives to embryo progress (blastocyst formation), time-lapse imaging data of preimplantation human embryo development were used. This retrospective study is focused on the topographic features and time parameters of the cleavages, with particular emphasis on the lengths of cleavage cycles and the genealogy of blastomeres in 2- to 8-cell human embryos. We have found that all 4-cell human embryos have four developmental variants that are based on the sequence of appearance and orientation of cleavage planes during embryo cleavage from 2 to 4 blastomeres. Each variant of cleavage shows a strong correlation with further developmental dynamics of the embryos (different cleavage cycle characteristics as well as lengths of blastomere cycles). An analysis of the sequence of human blastomere divisions allowed us to postulate that the effects of zygotic determinants are eliminated as a result of cleavage, and that, thereafter, blastomeres acquire the ability of own syntheses, regulation, polarization, formation of functional contacts, and, finally, of specific differentiation. This data on the early development of human embryos obtained using noninvasive methods complements and extend our understanding of the embryogenesis of eutherian mammals and may be applied in the practice of reproductive technologies.

scholarly journals Evaluating the utility of time-lapse imaging in the estimation of post-mortem interval: An Australian case study

2019 ◽  
Vol 1 ◽  
pp. 204-210 ◽  
Author(s):  
Alyson Wilson ◽  
Stanley Serafin ◽  
Dilan Seckiner ◽  
Rachel Berry ◽  
Xanthé Mallett
Keyword(s):  
Time Lapse ◽  
Post Mortem ◽  
Post Mortem Interval ◽  
Australian Case ◽  
Time Lapse Imaging
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