scholarly journals Light‐sheet microscopy‐based 3D single‐cell tracking reveals a correlation between cell cycle and the start of endoderm cell internalization in early zebrafish development

2020 ◽  
Vol 62 (7-8) ◽  
pp. 495-502
Author(s):  
Akiko Kondow ◽  
Kiyoshi Ohnuma ◽  
Yasuhiro Kamei ◽  
Atsushi Taniguchi ◽  
Ryoma Bise ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Single Cell ◽  
Cell Tracking ◽  
Light Sheet ◽  
Zebrafish Development ◽  
Light Sheet Microscopy ◽  
Endoderm Cell ◽  
Cell Internalization

scholarly journals Live-cell time-lapse imaging and single-cell tracking of in vitro cultured neural stem cells – Tools for analyzing dynamics of cell cycle, migration, and lineage selection

Methods ◽  
2018 ◽  
Vol 133 ◽  
pp. 81-90 ◽  
Author(s):  
Katja M. Piltti ◽  
Brian J. Cummings ◽  
Krystal Carta ◽  
Ayla Manughian-Peter ◽  
Colleen L. Worne ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Stem Cells ◽  
Neural Stem Cells ◽  
Single Cell ◽  
Cell Tracking ◽  
Time Lapse ◽  
Live Cell ◽  
Time Lapse Imaging

scholarly journals Optical tomography complements light sheet microscopy for in toto imaging of zebrafish development

Development ◽  
2015 ◽  
Vol 142 (5) ◽  
pp. 1016-1020 ◽  
Author(s):  
A. Bassi ◽  
B. Schmid ◽  
J. Huisken
Keyword(s):  
Optical Tomography ◽  
Light Sheet ◽  
Zebrafish Development ◽  
Light Sheet Microscopy

scholarly journals Automated cell cycle and cell size measurements for single-cell gene expression studies

2017 ◽  
Author(s):  
Anissa Guillemin ◽  
Angelique Richard ◽  
Sandrine Gonin-Giraud ◽  
Olivier Gandrillon
Keyword(s):  
Gene Expression ◽  
Cell Cycle ◽  
Single Cell ◽  
Cell Size ◽  
Cell Tracking ◽  
Erythroid Progenitors ◽  
Expression Variability ◽  
Unbiased Gene ◽  
Transcriptomic Level ◽  
Cell To Cell Variability

AbstractRecent rise of single-cell studies revealed the importance of understanding the role of cell-to-cell variability, especially at the transcriptomic level. One of the numerous sources of cell-to-cell variation in gene expression is the heterogeneity in cell proliferation state. How cell cycle and cell size influences gene expression variability at single-cell level is not yet clearly understood. To deconvolute such influences, most of the single-cell studies used dedicated methods that could include some bias. Here, we provide a universal and automatic toxic-free label method, compatible with single-cell high-throughput RT-qPCR. This led to an unbiased gene expression analysis and could be also used for improving single-cell tracking and imaging when combined with cell isolation. As an application for this technique, we showed that cell-to-cell variability in chicken erythroid progenitors was negligibly influenced by cell size nor cell cycle.

scholarly journals Visualizing Whole-Brain Activity and Development at the Single-Cell Level Using Light-Sheet Microscopy

Neuron ◽  
2015 ◽  
Vol 85 (3) ◽  
pp. 462-483 ◽  
Author(s):  
Philipp J. Keller ◽  
Misha B. Ahrens
Keyword(s):  
Single Cell ◽  
Brain Activity ◽  
Light Sheet ◽  
Single Cell Level ◽  
Cell Level ◽  
Whole Brain ◽  
Light Sheet Microscopy

scholarly journals 3D projection electrophoresis for single-cell immunoblotting

2019 ◽  
Author(s):  
Samantha M. Grist ◽  
Andoni P. Mourdoukoutas ◽  
Amy E. Herr
Keyword(s):  
Single Cell ◽  
Three Dimensional ◽  
Light Sheet ◽  
Single Cell Protein ◽  
Design Guidelines ◽  
Cell Protein ◽  
Separation Performance ◽  
The Novel ◽  
Light Sheet Microscopy ◽  
Serial Sampling

AbstractWhile immunoassays and mass spectrometry are powerful single-cell protein analysis tools, bottlenecks remain in interfacing and throughput. Here, we introduce highly parallel, synchronous, three-dimensional single-cell immunoblots to detect both cytosolic and nuclear proteins. The novel threedimensional microfluidic device is a photoactive polyacrylamide gel with a high-density microwell array patterned on one face (x-y) for cell isolation and lysis. From each microwell, single-cell lysate is ‘electrophoretically projected’ into the 3rd dimension (z-axis), separated by size, and photo-captured for immunoprobing and three-dimensional interrogation by confocal/light sheet microscopy. Design guidelines for throughput and separation performance are informed by simulation, analyses, and deconvolution postprocessing based on physics of 3D diffusion. Importantly, separations are nearly synchronous, whereas serial analyses can impart hours of delay between the first and last cell. We achieve an electrophoresis throughput of >2.5 cells/s (70X faster than serial sampling) and perform 25 immunoblots/mm2 device area (>10X increase over previous immunoblots). A straightforward device for parallel single-cell immunoblotting, projection electrophoresis promises to advance integration of protein-level profiles into the emerging single-cell atlas of genomic and transcriptomic profiles.

scholarly journals Computationally Enhanced Quantitative Phase Microscopy Reveals Autonomous Oscillations in Mammalian Cell Growth

2019 ◽  
Author(s):  
Xili Liu ◽  
Seungeun Oh ◽  
Leonid Peshkin ◽  
Marc W. Kirschner
Keyword(s):  
Cell Cycle ◽  
Growth Rate ◽  
Cell Growth ◽  
Single Cell ◽  
Cell Lines ◽  
Mammalian Cells ◽  
Cell Tracking ◽  
Quantitative Phase ◽  
Phase Microscopy ◽  
Quantitative Phase Microscopy

AbstractThe fine balance of growth and division is a fundamental property of the physiology of cells and one of the least understood. Its study has been thwarted by difficulties in the accurate measurement of cell size and the even greater challenges of measuring growth of a single-cell over time. We address these limitations by demonstrating a new computationally enhanced methodology for Quantitative Phase Microscopy (ceQPM) for adherent cells, using improved image processing algorithms and automated cell tracking software. Accuracy has been improved more than two-fold and this improvement is sufficient to establish the dynamics of cell growth and adherence to simple growth laws. It is also sufficient to reveal unknown features of cell growth previously unmeasurable. With these methodological and analytical improvements, we document a remarkable oscillation in growth rate in several different cell lines, occurring throughout the cell cycle, coupled to cell division or birth, and yet independent of cell cycle progression. We expect that further exploration with this improved tool will provide a better understanding of growth rate regulation in mammalian cells.Significance StatementIt has been a long-standing question in cell growth studies that whether the mass of individual cell grows linearly or exponentially. The two models imply fundamentally distinct mechanisms, and the discrimination of the two requires great measurement accuracy. Here, we develop a new method of computationally enhanced Quantitative Phase Microscopy (ceQPM), which greatly improves the accuracy and throughput of single-cell growth measurement in adherent mammalian cells. The measurements of several cell lines indicate that the growth dynamics of individual cells cannot be explained by either of the simple models but rather present an unanticipated and remarkable oscillatory behavior, suggesting more complex regulation and feedbacks.

scholarly journals Reconstruction of cell lineages and behaviors underlying arthropod limb outgrowth with multi-view light-sheet imaging and tracking

2017 ◽  
Author(s):  
Carsten Wolff ◽  
Jean-Yves Tinevez ◽  
Tobias Pietzsch ◽  
Evangelia Stamataki ◽  
Benjamin Harich ◽  
...  
Keyword(s):  
Cell Proliferation ◽  
Single Cell ◽  
Limb Development ◽  
Light Sheet ◽  
Limb Bud ◽  
List Type ◽  
Cell Behaviors ◽  
Light Sheet Microscopy ◽  
Organ Morphogenesis ◽  
Parhyale Hawaiensis

SUMMARYDuring development coordinated cell behaviors orchestrate tissue and organ morphogenesis to suit the lifestyle of the organism. We have used here the crustacean Parhyale hawaiensis to study the cellular basis of limb development. Transgenic Parhyale embryos with fluorescently labeled nuclei were imaged at high spatiotemporal resolution with multi-view light-sheet fluorescence microscopy over several days of embryogenesis spanning appendage morphogenesis from early specification up to late differentiation stages. Cell tracking with a new tool called Massive Multi-view Tracker (MaMuT) enabled the reconstruction of the complete cell lineage of an outgrowing thoracic limb with single-cell resolution. In silico clonal analyses suggested that the limb primordium becomes subdivided from an early stage first into anterior-posterior and then into dorsal-ventral compartments whose boundaries intersect at the distal tip of the growing limb. Limb bud formation is associated with the spatial modulation of cell proliferation, while limb elongation is also driven by the preferential orientation of division of epidermal cells along the proximal-distal axis of growth. Cellular reconstructions were predictive of the expression patterns of limb development genes including the Decapentaplegic (Dpp) morphogen.HIGHLIGHTSMulti-view light-sheet microscopy of crustacean embryos from species Parhyale hawaiensis are ideal for cellular-level analysis of organ morphogenesis.Lineages of 3-dimensional organs were reconstructed at single-cell resolution with the Fiji/ImageJ plugin Massive Multi-view Tracker.The Parhyale limb primordium undergoes early lineage restrictions associated with particular cell behaviors and patterns of gene expression.Differential rates of cell proliferation and oriented cell divisions guide appendage proximal-distal outgrowth.

scholarly journals Recent Progress in Light Sheet Microscopy for Biological Applications

2018 ◽  
Vol 72 (8) ◽  
pp. 1137-1169 ◽  
Author(s):  
Krishnendu Chatterjee ◽  
Feby Wijaya Pratiwi ◽  
Frances Camille M. Wu ◽  
Peilin Chen ◽  
Bi-Chang Chen
Keyword(s):  
Developmental Biology ◽  
Fluorescence Microscopy ◽  
Single Cell ◽  
Single Molecule ◽  
Super Resolution ◽  
Light Sheet ◽  
High Signal ◽  
Spatiotemporal Resolution ◽  
Light Sheet Microscopy ◽  
Resolution Imaging

The introduction of light sheet fluorescence microscopy (LSFM) has overcome the challenges in conventional optical microscopy. Among the recent breakthroughs in fluorescence microscopy, LSFM had been proven to provide a high three-dimensional spatial resolution, high signal-to-noise ratio, fast imaging acquisition rate, and minuscule levels of phototoxic and photodamage effects. The aforementioned auspicious properties are crucial in the biomedical and clinical research fields, covering a broad range of applications: from the super-resolution imaging of intracellular dynamics in a single cell to the high spatiotemporal resolution imaging of developmental dynamics in an entirely large organism. In this review, we provided a systematic outline of the historical development of LSFM, detailed discussion on the variants and improvements of LSFM, and delineation on the most recent technological advancements of LSFM and its potential applications in single molecule/particle detection, single-molecule super-resolution imaging, imaging intracellular dynamics of a single cell, multicellular imaging: cell–cell and cell–matrix interactions, plant developmental biology, and brain imaging and developmental biology.

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