scholarly journals Multi-modal in vivo visualization of single cell dynamics by 1P, 2P, light-sheet, and on-chip technologies

2016 ◽  
pp. 147-148
Author(s):  
Satoshi Nishimura
Keyword(s):  
Single Cell ◽  
Light Sheet ◽  
Cell Dynamics ◽  
On Chip

scholarly journals Stochastic Single Cell Behaviour Leads to Robust Horizontal Cell Layer Formation in the Vertebrate Retina

2019 ◽  
Author(s):  
Rana Amini ◽  
Anastasia Labudina ◽  
Caren Norden
Keyword(s):  
Single Cell ◽  
Cell Layer ◽  
Horizontal Cell ◽  
Light Sheet ◽  
Cell Dynamics ◽  
Cell Behaviour ◽  
Vertebrate Retina ◽  
Laminar Pattern ◽  
Cell Cycle Dynamics

ABSTRACTDevelopmental programs that arrange cells and tissues into patterned organs are remarkably robust. In the developing vertebrate retina for example, neurons reproducibly assemble into distinct layers giving the mature organ its overall structured appearance. This stereotypic neuronal arrangement, termed lamination, is important for efficient neuronal connectivity. While retinal lamination is conserved in many vertebrates including humans, how it emerges from single cell behaviour is not fully understood. To shed light on this question, we here investigated the formation of the retinal horizontal cell layer. Using in vivo light sheet imaging of the developing zebrafish retina, we generated a comprehensive quantitative analysis of horizontal single cell behaviour from birth to final positioning. Interestingly, we find that all parameters analyzed including cell cycle dynamics, migration paths and kinetics as well as sister cell dispersal are very heterogeneous. Thus, horizontal cells show individual non-stereotypic behaviour before final positioning. Yet, these initially stochastic cell dynamics always generate the correct laminar pattern. Consequently, our data shows that lamination of the vertebrate retina contains a yet underexplored extent of single cell stochasticity.

scholarly journals In Vivo Biochemistry: Single-Cell Dynamics of Cyclic Di-GMP in Escherichia coli in Response to Zinc Overload

Biochemistry ◽  
2017 ◽  
Vol 57 (1) ◽  
pp. 108-116 ◽  
Author(s):  
Jongchan Yeo ◽  
Andrew B. Dippel ◽  
Xin C. Wang ◽  
Ming C. Hammond
Keyword(s):  
Escherichia Coli ◽  
Single Cell ◽  
Cell Dynamics

scholarly journals Single-cell transcriptomics elucidates in vitro reprogramming of human intestinal epithelium cultured in a physiodynamic gut-on-a-chip

2021 ◽  
Author(s):  
Woojung Shin ◽  
Zhe Su ◽  
S. Stephen Yi ◽  
Hyun Jung Kim
Keyword(s):  
Single Cell ◽  
Intestinal Epithelium ◽  
Drug Transport ◽  
Three Dimensional ◽  
Cancer Cell Line ◽  
Static Condition ◽  
On Chip ◽  
Epithelial Layers

The microphysiological human gut-on-a-chip has demonstrated in vivo-relevant cellular fidelity of intestinal epithelium compared to its cultures in a static condition. Microfluidic control of morphogen gradients and mechanical cues robustly induced morphological histogenesis with villi-like three-dimensional (3D) microarchitecture, lineage-associated cytodifferentiation, and physiological functions of a human intestinal Caco-2 epithelium. However, transcriptomic dynamics that orchestrates morphological and functional reprogramming of the epithelium in a microphysiological culture remains elusive. Single-cell transcriptomic analysis revealed that a gut-on-a-chip culture that offers physiological motions and flow drives three distinctive subclusters that offer distinct gene expression and unique spatial representation in 3D epithelial layers. The pseudotemporal trajectory of individual cells visualized the evolutionary transition from ancestral genotypes in static cultures into more heterogeneous phenotypes in physiodynamic cultures on cell cycles, differentiation, and intestinal functions including digestion, absorption, drug transport, and metabolism of xenobiotics. Furthermore, the inversed transcriptomic signature of oncogenes and tumor-suppressor genes of Caco-2 cells verified that a gut-on-a-chip culture drives a postmitotic reprogramming of cancer-associated phenotypes. Thus, we discovered that a physiodynamic on-chip culture is necessary and sufficient for a cancer cell line to be reprogrammed to elicit in vivo-relevant heterogeneous cell populations with restored normal physiological signatures.

scholarly journals Zebrafish Microenvironment Elevates EMT and CSC-Like Phenotype of Engrafted Prostate Cancer Cells

Cells ◽  
2020 ◽  
Vol 9 (4) ◽  
pp. 797
Author(s):  
Lanpeng Chen ◽  
Maciej Boleslaw Olszewski ◽  
Marianna Kruithof-de Julio ◽  
B. Ewa Snaar-Jagalska
Keyword(s):  
Prostate Cancer ◽  
Single Cell ◽  
Epithelial Mesenchymal Transition ◽  
Early Stage ◽  
Xenograft Model ◽  
Light Sheet ◽  
Hematopoietic Tissue ◽  
Cell Dynamics ◽  
Mesenchymal Transition ◽  
Cell Subpopulations

To visually and genetically trace single-cell dynamics of human prostate cancer (PCa) cells at the early stage of metastasis, a zebrafish (ZF) xenograft model was employed. The phenotypes of intravenously transplanted fluorescent cells were monitored by high-resolution, single-cell intravital confocal and light-sheet imaging. Engrafted osteotropic, androgen independent PCa cells were extravasated from caudle vein, invaded the neighboring tissue, proliferated and formed experimental metastases around caudal hematopoietic tissue (CHT) in four days. Gene expression comparison between cells in culture and in CHT revealed that engrafted PCa cells responded to the ZF microenvironment by elevating expression of epithelial–mesenchymal transition (EMT) and stemness markers. Next, metastatic potentials of ALDHhi cancer stem-like cells (CSCs) and ALDHlow non-CSCs were analyzed in ZF. Engraftment of CSCs induced faster metastatic onset, however after six days both cell subpopulations equally responded to the ZF microenvironment, resulting in the same increase of stemness genes expression including Nanog, Oct-4 and Cripto. Knockdown of Cripto significantly reduced the vimentin/E-cadherin ratio in engrafted cells, indicating that Cripto is required for transduction of the microenvironment signals from the ZF niche to increase mesenchymal potential of cells. Targeting of either Cripto or EMT transcriptional factors Snail 1 and Zeb1 significantly suppressed metastatic growth. These data indicated that zebrafish microenvironment governed the CSC/EMT plasticity of human PCa cells promoting metastasis initiation.

scholarly journals Imaging the Single Cell Dynamics of CD4+ T Cell Activation by Dendritic Cells in Lymph Nodes

2004 ◽  
Vol 200 (7) ◽  
pp. 847-856 ◽  
Author(s):  
Mark J. Miller ◽  
Olga Safrina ◽  
Ian Parker ◽  
Michael D. Cahalan
Keyword(s):  
T Cells ◽  
Dendritic Cells ◽  
T Cell ◽  
Lymph Nodes ◽  
Single Cell ◽  
Cd4 T Cells ◽  
Antigen Recognition ◽  
Time Behavior ◽  
Cell Dynamics

The adaptive immune response is initiated in secondary lymphoid organs by contact between antigen-bearing dendritic cells (DCs) and antigen-specific CD4+ T cells. However, there is scant information regarding the single cell dynamics of this process in vivo. Using two-photon microscopy, we imaged the real-time behavior of naive CD4+ T cells and in vivo–labeled DCs in lymph nodes during a robust T cell response. In the first 2 h after entry into lymph nodes, T cells made short-lived contacts with antigen-bearing DCs, each contact lasting an average of 11–12 min and occurring mainly on dendrites. Altered patterns of T cell motility during this early stage of antigen recognition promoted serial engagement with several adjacent DCs. Subsequently, T cell behavior progressed through additional distinct stages, including long-lived clusters, dynamic swarms, and finally autonomous migration punctuated by cell division. These observations suggest that the immunological synapse in native tissues is remarkably fluid, and that stable synapses form only at specific stages of antigen presentation to T cells. Furthermore, the serial nature of these interactions implies that T cells activate by way of multiple antigen recognition events.

Raman spectroscopy compatible PDMS droplet microfluidic culture and analysis platform towards on-chip lipidomics

The Analyst ◽  
2017 ◽  
Vol 142 (7) ◽  
pp. 1054-1060 ◽  
Author(s):  
Hyun Soo Kim ◽  
Sergio C. Waqued ◽  
Dawson T. Nodurft ◽  
Timothy P. Devarenne ◽  
Vladislav V. Yakovlev ◽  
...  
Keyword(s):  
Raman Spectroscopy ◽  
Single Cell ◽  
Lipid Analysis ◽  
On Chip ◽  
Analysis Platform ◽  
Biomolecular Analysis

We present a method that allows for the use of Raman spectroscopy with PDMS-based microdevices to perform on-chip, droplet-based in vivo biomolecular analysis (i.e., microalgal lipid analysis) with single-cell resolution.

scholarly journals In vivo NIR-II structured-illumination light-sheet microscopy

2021 ◽  
Vol 118 (6) ◽  
pp. e2023888118
Author(s):  
Feifei Wang ◽  
Zhuoran Ma ◽  
Yeteng Zhong ◽  
Felix Salazar ◽  
Chun Xu ◽  
...  
Keyword(s):  
Light Scattering ◽  
Molecular Imaging ◽  
Single Cell ◽  
Light Sheet ◽  
Single Cell Level ◽  
Structured Illumination ◽  
Cell Level ◽  
Spatiotemporal Resolution ◽  
Light Sheet Microscopy

Noninvasive optical imaging with deep tissue penetration depth and high spatiotemporal resolution is important to longitudinally studying the biology at the single-cell level in live mammals, but has been challenging due to light scattering. Here, we developed near-infrared II (NIR-II) (1,000 to 1,700 nm) structured-illumination light-sheet microscopy (NIR-II SIM) with ultralong excitation and emission wavelengths up to ∼1,540 and ∼1,700 nm, respectively, suppressing light scattering to afford large volumetric three-dimensional (3D) imaging of tissues with deep-axial penetration depths. Integrating structured illumination into NIR-II light-sheet microscopy further diminished background and improved spatial resolution by approximately twofold. In vivo oblique NIR-II SIM was performed noninvasively for 3D volumetric multiplexed molecular imaging of the CT26 tumor microenvironment in mice, longitudinally mapping out CD4, CD8, and OX40 at the single-cell level in response to immunotherapy by cytosine-phosphate-guanine (CpG), a Toll-like receptor 9 (TLR-9) agonist combined with OX40 antibody treatment. NIR-II SIM affords an additional tool for noninvasive volumetric molecular imaging of immune cells in live mammals.

scholarly journals Emergence of single cell mechanical behavior and polarity within epithelial monolayers drives collective cell migration

2019 ◽  
Author(s):  
Shreyansh Jain ◽  
Victoire M.L. Cachoux ◽  
Gautham H.N.S. Narayana ◽  
Simon de Beco ◽  
Joseph D’Alessandro ◽  
...  
Keyword(s):  
Cell Migration ◽  
Single Cell ◽  
Large Scale ◽  
Cancer Metastasis ◽  
Cell Junctions ◽  
Collective Cell Migration ◽  
Convergent Extension ◽  
Cell Dynamics

The directed migration of cell collectives is essential in various physiological processes, such as epiboly, intestinal epithelial turnover, and convergent extension during morphogenesis as well as during pathological events like wound healing and cancer metastasis1,2. Collective cell migration leads to the emergence of coordinated movements over multiple cells. Our current understanding emphasizes that these movements are mainly driven by large-scale transmission of signals through adherens junctions3,4. In this study, we show that collective movements of epithelial cells can be triggered by polarity signals at the single cell level through the establishment of coordinated lamellipodial protrusions. We designed a minimalistic model system to generate one-dimensional epithelial trains confined in ring shaped patterns that recapitulate rotational movements observed in vitro in cellular monolayers and in vivo in genitalia or follicular cell rotation5–7. Using our system, we demonstrated that cells follow coordinated rotational movements after the establishment of directed Rac1-dependent polarity over the entire monolayer. Our experimental and numerical approaches show that the maintenance of coordinated migration requires the acquisition of a front-back polarity within each single cell but does not require the maintenance of cell-cell junctions. Taken together, these unexpected findings demonstrate that collective cell dynamics in closed environments as observed in multiple in vitro and in vivo situations5,6,8,9 can arise from single cell behavior through a sustained memory of cell polarity.

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