Autofocus System for Imaging Multiple Cells Across Thick Liquid Layers in Differrent Focal Planes

2020 ◽  
Author(s):  
Lin Liu ◽  
Xiaohui Du ◽  
Juanxiu Liu ◽  
Guangming Ni ◽  
Jing Zhang ◽  
...  
Keyword(s):  
Multiple Cells

scholarly journals Role of interleukins in the pathogenesis of pulmonary fibrosis

2021 ◽  
Vol 7 (1) ◽  
Author(s):  
Yi Xin She ◽  
Qing Yang Yu ◽  
Xiao Xiao Tang
Keyword(s):  
Pulmonary Fibrosis ◽  
Therapeutic Strategy ◽  
Future Directions ◽  
Regulation Mechanisms ◽  
Underlying Mechanisms ◽  
Multiple Cells ◽  
The Relationship

AbstractInterleukins, a group of cytokines participating in inflammation and immune response, are proved to be involved in the formation and development of pulmonary fibrosis. In this article, we reviewed the relationship between interleukins and pulmonary fibrosis from the clinical, animal, as well as cellular levels, and discussed the underlying mechanisms in vivo and in vitro. Despite the effects of interleukin-targeted treatment on experimental pulmonary fibrosis, clinical applications are lacking and unsatisfactory. We conclude that intervening in one type of interleukins with similar functions in IPF may not be enough to stop the development of fibrosis as it involves a complex network of regulation mechanisms. Intervening interleukins combined with other existing therapy or targeting interleukins affecting multiple cells/with different functions at the same time may be one of the future directions. Furthermore, the intervention time is critical as some interleukins play different roles at different stages. Further elucidation on these aspects would provide new perspectives on both the pathogenesis mechanism, as well as the therapeutic strategy and drug development.

scholarly journals G Protein-Coupled Estrogen Receptor in Cancer and Stromal Cells: Functions and Novel Therapeutic Perspectives

Cells ◽  
2021 ◽  
Vol 10 (3) ◽  
pp. 672
Author(s):  
Richard A. Pepermans ◽  
Geetanjali Sharma ◽  
Eric R. Prossnitz
Keyword(s):  
Estrogen Receptor ◽  
G Protein ◽  
Inflammatory Diseases ◽  
Therapy Resistance ◽  
Breast Cancers ◽  
Obesity And Diabetes ◽  
Therapeutic Value ◽  
Health And Disease ◽  
Multiple Cells ◽  
G Protein Coupled

Estrogen is involved in numerous physiological and pathophysiological systems. Its role in driving estrogen receptor-expressing breast cancers is well established, but it also has important roles in a number of other cancers, acting both on tumor cells directly as well as in the function of multiple cells of the tumor microenvironment, including fibroblasts, immune cells, and adipocytes, which can greatly impact carcinogenesis. One of its receptors, the G protein-coupled estrogen receptor (GPER), has gained much interest over the last decade in both health and disease. Increasing evidence shows that GPER contributes to clinically observed endocrine therapy resistance in breast cancer while also playing a complex role in a number of other cancers. Recent discoveries regarding the targeting of GPER in combination with immune checkpoint inhibition, particularly in melanoma, have led to the initiation of the first Phase I clinical trial for the GPER-selective agonist G-1. Furthermore, its functions in metabolism and corresponding pathophysiological states, such as obesity and diabetes, are becoming more evident and suggest additional therapeutic value in targeting GPER for both cancer and other diseases. Here, we highlight the roles of GPER in several cancers, as well as in metabolism and immune regulation, and discuss the therapeutic value of targeting this estrogen receptor as a potential treatment for cancer as well as contributing metabolic and inflammatory diseases and conditions.

scholarly journals C3VFC: A Method for Tracing and Quantification of Microglia in 3D Temporal Images

2021 ◽  
Vol 11 (13) ◽  
pp. 6078
Author(s):  
Tiffany T. Ly ◽  
Jie Wang ◽  
Kanchan Bisht ◽  
Ukpong Eyo ◽  
Scott T. Acton
Keyword(s):  
Vector Field ◽  
Critical Points ◽  
State Of The Art ◽  
Temporal Data ◽  
Entire Structure ◽  
Tracing Algorithm ◽  
Vector Field Convolution ◽  
Multiple Cells ◽  
Tracing Method ◽  
Over Time

Automatic glia reconstruction is essential for the dynamic analysis of microglia motility and morphology, notably so in research on neurodegenerative diseases. In this paper, we propose an automatic 3D tracing algorithm called C3VFC that uses vector field convolution to find the critical points along the centerline of an object and trace paths that traverse back to the soma of every cell in an image. The solution provides detection and labeling of multiple cells in an image over time, leading to multi-object reconstruction. The reconstruction results can be used to extract bioinformatics from temporal data in different settings. The C3VFC reconstruction results found up to a 53% improvement on the next best performing state-of-the-art tracing method. C3VFC achieved the highest accuracy scores, in relation to the baseline results, in four of the five different measures: Entire structure average, the average bi-directional entire structure average, the different structure average, and the percentage of different structures.

scholarly journals Multiphoton Imaging of Ca2+ Instability in Acute Myocardial Slices from a RyR2R2474S Murine Model of Catecholaminergic Polymorphic Ventricular Tachycardia

2021 ◽  
Vol 10 (13) ◽  
pp. 2821
Author(s):  
Giulia Borile ◽  
Tania Zaglia ◽  
Stephan E. Lehnart ◽  
Marco Mongillo
Keyword(s):  
Ventricular Tachycardia ◽  
Single Cells ◽  
Tissue Level ◽  
Catecholaminergic Polymorphic Ventricular Tachycardia ◽  
Polymorphic Ventricular Tachycardia ◽  
Adrenergic Stimulation ◽  
Release Channel ◽  
Multiphoton Imaging ◽  
Delayed Afterdepolarizations ◽  
Multiple Cells

Catecholaminergic Polymorphic Ventricular Tachycardia (CPVT) is a familial stress-induced arrhythmia syndrome, mostly caused by mutations in Ryanodine receptor 2 (RyR2), the sarcoplasmic reticulum (SR) Ca2+ release channel in cardiomyocytes. Pathogenetic mutations lead to gain of function in the channel, causing arrhythmias by promoting diastolic spontaneous Ca2+ release (SCR) from the SR and delayed afterdepolarizations. While the study of Ca2+ dynamics in single cells from murine CPVT models has increased our understanding of the disease pathogenesis, questions remain on the mechanisms triggering the lethal arrhythmias at tissue level. Here, we combined subcellular analysis of Ca2+ signals in isolated cardiomyocytes and in acute thick ventricular slices of RyR2R2474S knock-in mice, electrically paced at different rates (1–5 Hz), to identify arrhythmogenic Ca2+ dynamics, from the sub- to the multicellular perspective. In both models, RyR2R2474S cardiomyocytes had increased propensity to develop SCR upon adrenergic stimulation, which manifested, in the slices, with Ca2+ alternans and synchronous Ca2+ release events in neighboring cardiomyocytes. Analysis of Ca2+ dynamics in multiple cells in the tissue suggests that SCRs beget SCRs in contiguous cells, overcoming the protective electrotonic myocardial coupling, and potentially generating arrhythmia triggering foci. We suggest that intercellular interactions may underscore arrhythmic propensity in CPVT hearts with ‘leaky’ RyR2.

scholarly journals Insect Wing Membrane Topography Is Determined by the Dorsal Wing Epithelium

2013 ◽  
Vol 3 (1) ◽  
pp. 5-8 ◽  
Author(s):  
Andrea D Belalcazar ◽  
Kristy Doyle ◽  
Justin Hogan ◽  
David Neff ◽  
Simon Collier
Keyword(s):  
Planar Cell Polarity ◽  
Parasitic Wasp ◽  
Ventral Surface ◽  
Wing Membrane ◽  
Genetic Studies ◽  
Insect Wing ◽  
Membrane Topography ◽  
Wing Hair ◽  
Pcp Signaling ◽  
Multiple Cells

Abstract The Drosophila wing consists of a transparent wing membrane supported by a network of wing veins. Previously, we have shown that the wing membrane cuticle is not flat but is organized into ridges that are the equivalent of one wing epithelial cell in width and multiple cells in length. These cuticle ridges have an anteroposterior orientation in the anterior wing and a proximodistal orientation in the posterior wing. The precise topography of the wing membrane is remarkable because it is a fusion of two independent cuticle contributions from the dorsal and ventral wing epithelia. Here, through morphological and genetic studies, we show that it is the dorsal wing epithelium that determines wing membrane topography. Specifically, we find that wing hair location and membrane topography are coordinated on the dorsal, but not ventral, surface of the wing. In addition, we find that altering Frizzled Planar Cell Polarity (i.e., Fz PCP) signaling in the dorsal wing epithelium alone changes the membrane topography of both dorsal and ventral wing surfaces. We also examined the wing morphology of two model Hymenopterans, the honeybee Apis mellifera and the parasitic wasp Nasonia vitripennis. In both cases, wing hair location and wing membrane topography are coordinated on the dorsal, but not ventral, wing surface, suggesting that the dorsal wing epithelium also controls wing topography in these species. Because phylogenomic studies have identified the Hymenotera as basal within the Endopterygota family tree, these findings suggest that this is a primitive insect character.

Development of a carboxyl-terminated Indium Tin Oxide electrode for improving cell adhesion and facilitating low noise, real-time impedance measurements

2021 ◽  
Author(s):  
A.M. Ilyas ◽  
Md Kowsar Alam ◽  
Jamal-Deen Musah ◽  
Lin Oo SAW ◽  
Chi-Chung Yeung ◽  
...  
Keyword(s):  
Cell Adhesion ◽  
Tin Oxide ◽  
Indium Tin Oxide ◽  
Time Course ◽  
Low Noise ◽  
Collective Cell Migration ◽  
Signal Loss ◽  
Ito Cell ◽  
Cell Substrate ◽  
Multiple Cells

The working electrode's surface property is crucial to cell adhesion and signal collection in electric cell-substrate impedance spectroscopy (ECIS). To date, the indium tin oxide (ITO) based working electrode is of interest in ECIS study due to its high transparency and biocompatibility. Of great concern is the impedance signal loss, distortion, and data interpretation conflict profoundly created by the movement of multiple cells during ECIS study. Here, a carboxyl- terminated-ITO substrate was prepared by stepwise surface amino silanization, with N-Hydroxy succinimide (NHS), and (1-ethyl-3-(3-dimethyl aminopropyl) carbodiimide hydrochloride) (EDC) treatment, respectively. We investigated the stepwise changes in the property of the treated ITO, cell-substrate adhesion, collective cell mobility, and time course of change in absolute impedance from multiple CHO cells ((Δt-Δ|Z|)CELLS). The carboxyl-terminated ITO substrate with a surface roughness of 6.37 nm shows enhanced conductivity, 75% visible light transparency, improved cell adherence, reduced collective cell migration speed by ~2 fold, and diminished signal distortion in the ((Δt-Δ|Z|)CELLS). Thus, our study provides an ITO surface-treatment strategy to reduce multiple cell movement effects and to obtain essential cell information from the ECIS study of multiple cells through undistorted (Δt-Δ|Z|)CELLS

scholarly journals U-shaped Dose Response in Vasomotor Tone: A Mixed Result of Heterogenic Response of Multiple Cells to Xenobiotics

2008 ◽  
Vol 103 (1) ◽  
pp. 181-190 ◽  
Author(s):  
Ok-Nam Bae ◽  
Kyung-Min Lim ◽  
Jee-Yeon Han ◽  
Byoung-In Jung ◽  
Jin-Young Lee ◽  
...  
Keyword(s):  
Dose Response ◽  
Mixed Result ◽  
Vasomotor Tone ◽  
Multiple Cells

scholarly journals Defining the early stages of intestinal colonisation by whipworms

2020 ◽  
Author(s):  
María A. Duque-Correa ◽  
David Goulding ◽  
Claire Cormie ◽  
Catherine Sharpe ◽  
Judit Gali Moya ◽  
...  
Keyword(s):  
Intestinal Epithelium ◽  
Early Stage ◽  
Proximal Colon ◽  
Host Cells ◽  
Parasitic Nematodes ◽  
Metazoan Parasites ◽  
Multiple Cells ◽  
First Time

ABSTRACTHundreds of millions of people are infected with whipworms (Trichuris trichiura), large metazoan parasites that live in the caecum and proximal colon. Whipworms inhabit distinct multi-intracellular epithelial burrows that have been described as syncytial tunnels. However, the interactions between first-stage (L1) larvae and the host epithelia that determine parasite invasion and establishment in the syncytium remain unclear. In vivo experiments investigating these events have been severely hampered by the limited in situ accessibility to intracellular infective larvae at the bottom of the crypts of Lieberkühn, and the lack of genetic tools such as fluorescent organisms that are readily available for other pathogens but not parasitic nematodes. Moreover, cell lines, which do not mimic the complexity of the intestinal epithelium, have been unsuccessful in supporting infection by whipworm larvae. Here, we show that caecaloids grown in an open crypt-like conformation recapitulate the caecal epithelium. Using this system, we establish in vitro infections with T. muris L1 larvae for the first-time, and provide clear evidence that syncytial tunnels are formed at this early stage. We show that larval whipworms are completely intracellular but woven through multiple cells. Using the caecaloids, we are able to visualise the pathways taken by the larvae as they burrow through the epithelial cells. We also demonstrate that larvae degrade the mucus layers overlaying the epithelium, enabling them to access the cells below. We show that early syncytial tunnels are composed of enterocytes and goblet cells that are alive and actively interacting with the larvae during the first 24 h of the infection. Progression of infection results in damage to host cells and by 72 h post-infection, we show that desmosomes of cells from infected epithelium widen and some host cells appear to become liquified. Collectively, our work unravels processes mediating the intestinal epithelium invasion by whipworms and reveals new specific interactions between the host and the parasite that allow the whipworm to establish on its multi-intracellular niche. Our study demonstrates that caecaloids can be used as a relevant in vitro model to investigate the infection biology of T. muris during the early colonisation of its host.

scholarly journals All-optical electrophysiology with improved genetically encoded voltage indicators reveals interneuron network dynamics in vivo

2021 ◽  
Author(s):  
He Tian ◽  
Hunter C. Davis ◽  
J. David Wong-Campos ◽  
Linlin Z. Fan ◽  
Benjamin Gmeiner ◽  
...  
Keyword(s):  
Gap Junction ◽  
Signal To Noise Ratio ◽  
Voltage Imaging ◽  
All Optical ◽  
Precision Voltage ◽  
Coupling Strengths ◽  
Junction Coupling ◽  
Multiple Cells ◽  
Voltage Indicator

All-optical electrophysiology can be a powerful tool for studying neural dynamics in vivo, as it offers the ability to image and perturb membrane voltage in multiple cells simultaneously. The "Optopatch" constructs combine a red-shifted archaerhodopsin (Arch)-derived genetically encoded voltage indicator (GEVI) with a blue-shifted channelrhodopsin actuator (ChR). We used a video-based pooled screen to evolve Arch-derived GEVIs with improved signal-to-noise ratio (QuasAr6a) and kinetics (QuasAr6b). By combining optogenetic stimulation of individual cells with high-precision voltage imaging in neighboring cells, we mapped inhibitory and gap junction-mediated connections, in vivo. Optogenetic activation of a single NDNF-expressing neuron in visual cortex Layer 1 significantly suppressed the spike rate in some neighboring NDNF interneurons. Hippocampal PV cells showed near-synchronous spikes across multiple cells at a frequency significantly above what one would expect from independent spiking, suggesting that collective inhibitory spikes may play an important signaling role in vivo. By stimulating individual cells and recording from neighbors, we quantified gap junction coupling strengths. Together, these results demonstrate powerful new tools for all-optical microcircuit dissection in live mice.

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