Reflections on Quantitative Gamma Imaging of Cell-Surface Interactions

Author(s):  
Robert C. Eberhart

Molecular and cellular interactions with foreign surfaces can be noninvasively measured by isotope imaging techniques. Long available for probing cell behavior, these techniques are now employed in molecular studies of disease progression, such as Alzheimer’s [1]. This paper reviews results obtained by noninvasive dual label gamma scintigraphy for the transient adhesion of platelets and neutrophils to pump-oxygenators during cardiopulmonary bypass (CPB). In this application, characteristic cell-foreign surface adhesion and release patterns are observed during CPB in the pig, as a function of oxygenator design and surface chemistry. Cell distributions in internal organs post-CPB are also affected by these processes. This method can be adapted to other settings where the understanding of protein-cell interactions with native and foreign surfaces is at issue, including fibrinogen-cell interactions, bacterial colonization, etc.

Development ◽  
1995 ◽  
Vol 121 (7) ◽  
pp. 2219-2232 ◽  
Author(s):  
R. Schnabel

During the first four cleavage rounds of the Caenorhabditis elegans embryo, five somatic founder cells AB, MS, E, C and D are born, which later form the tissues of the embryo. The classical criterion for a cell-autonomous specification of a tissue is the capability of primordial cells to produce this tissue in isolation from the remainder of the embryo. By this criterion, the somatic founder cells MS, C and D develop cell-autonomously. Laser ablation experiments, however, reveal that within the embryonic context these blastomeres form a network of duelling cellular interactions. During normal development, the blastomere D inhibits muscle specification in the MS and the C lineage inhibits muscle specification in the D lineage. These inhibitory interactions are counteracted by two activating inductions. As described before the inhibition of body wall muscle in MS is counteracted by an activating signal from the ABa lineage. Body wall muscle in the D lineage is induced by MS descendants, which suppress an inhibitory activity of the C lineage. The interaction between the D and the MS lineage occurs through the C lineage. An interesting feature of these cell-cell interactions is that they do not serve to discriminate between equivalent cells but are permissive or nonpermissive inductions. No evidence was found that the C-derived body wall muscle also depends on an induction, which suggests that possibly three different pathways coexist in the early embryo to specify body wall muscle, two of which are, in different ways, influenced by cell-cell interactions and a third that is autonomous. This work supplies evidence that cells may acquire transient states during embryogenesis that influence the specification of other cells in the embryo. These states, however, may not be reflected in the developmental potentials of the cells themselves. They can only be scored indirectly by their action on the specification of other cells in the embryo. Blastomeres that behave cell-autonomously in isolation are nevertheless subjected to cell-cell interactions in the embryonic context. Why this should be is an intriguing question. The classical notion has been that blastomeres are specified autonomously in nematodes. In recent years, it was established that at least five inductions are required to determine the AB descendants of C. elegans, whereas the P1 descendants have been typically viewed to develop more autonomously. It appears now that inductions also play a major role during the determination of P1-derived blastomeres.


2021 ◽  
Author(s):  
Rob C. Oslund ◽  
Tamara Reyes-Robles ◽  
Cory H. White ◽  
Jake H. Tomlinson ◽  
Kelly A. Crotty ◽  
...  

AbstractCell-cell interactions drive essential biological processes critical to cell and tissue development, function, pathology, and disease outcome. The growing appreciation of immune cell interactions within disease environments has led to significant efforts to develop protein- and cell-based therapeutic strategies. A better understanding of these cell-cell interactions will enable the development of effective immunotherapies. However, characterizing these complex cellular interactions at molecular resolution in their native biological contexts remains challenging. To address this, we introduce photocatalytic cell tagging (PhoTag), a modality agnostic platform for profiling cell-cell interactions. Using photoactivatable flavin-based cofactors, we generate phenoxy radical tags for targeted labeling at the cell surface. Through various targeting modalities (e.g. MHC-Multimer, antibody, single domain antibody (VHH)) we deliver a flavin photocatalyst for cell tagging within monoculture, co-culture, and peripheral blood mononuclear cells. PhoTag enables highly selective tagging of the immune synapse between an immune cell and an antigen-presenting cell through targeted labeling at the cell-cell junction. This allowed for the ability to profile gene expression-level differences between interacting and bystander cell populations. Given the modality agnostic and spatio-temporal nature of PhoTag, we envision its broad utilization to detect and profile intercellular interactions within an immune synapse and other confined cellular regions for any biological system.


Author(s):  
Giancarlo Ferrigno ◽  
Antonio Pedotti

Computer Aided Surgery is opening new scenarios in medicine aimed to improve the patient’s care by advancing the utilisation of computer into surgical practice which have already provided practical application particularly in orthopaedics and neurosurgery. The recent developments in multimodal 3D medical imaging techniques which are providing with great accuracy and the necessary resolution a clear representation on a multiscale basis of the morphological and functional features of the internal organs and tissues constitute the necessary basis. The interdisciplinary collaboration between engineers and physicians provide new concepts and applications in this field derived from digital modelling and simulation methods for the analysis of complex systems, sensors and motion capture technologies, robotics and mechatronics. Three main phases are described in this paper. The preoperative phase where the information derived from multimodal imaging of each patient is processed by proper mathematical models and simulation techniques to identify the best way to operate (surgical planning). The intraoperative phase where the surgeon is assisted by a “virtual fusion” of the preoperative images of the internal organs of the patients with the effective position of operative instruments recorded by proper trackers. Finally the postoperative phase to evaluate the efficacy of the treatments and the patient’s follow up.


1979 ◽  
Vol 39 (1) ◽  
pp. 201-213
Author(s):  
PIERANGELO LUPORINI ◽  
PAOLO BRACCHI ◽  
FULVIO ESPOSITO

A system has been developed to study cellular interactions between cells of complementary mating types prior to mating in the ciliate, Euplotes crassus. The presumptive mates were distinguished by using singlet and doublet cells of appropriate mating types in the mixtures. Cells of a given mating type were prelabelled with [3H]leucine and mixed with unlabelled complementary cells. Exchange of [3H]leucine-labelled material from donor to recipient cells was monitored through the various stages of the preconjugant interaction. A label transfer between the mating type complementary cells was detected from the beginning of the visible mating reaction, which occurs after a waiting period from the time of cell mixing and involves ciliary agglutination prior to cell body fusion. Complementary cells which were prevented from physically contacting each other and cells which were not competent to mate appeared unable to take up the labelled material. It is suggested that this material consists of some substance(s) playing an important role in the preconjugant cell-to-cell interactions of E. crassus.


2021 ◽  
Author(s):  
Yin Liu ◽  
Alex J Diaz de Arce ◽  
Mark A Krasnow

Interoceptors, sensory neurons that monitor internal organs and states, are essential for physiological homeostasis and generating internal perceptions. Here we describe a comprehensive transcriptomic atlas of interoceptors of the mouse lung, defining 10 molecular subtypes that differ in developmental origin, myelination, receptive fields, terminal morphologies, and cell contacts. Each subtype expresses a unique but overlapping combination of sensory receptors that detect diverse physiological and pathological stimuli, and each can signal to distinct sets of lung cells including immune cells, forming a local neuroimmune interaction network. Functional interrogation of two mechanosensory subtypes reveals exquisitely-specific homeostatic roles in breathing, one regulating inspiratory time and the other inspiratory flow. The results suggest that lung interoceptors encode diverse and dynamic sensory information rivaling that of canonical exteroceptors, and this information is used to drive myriad local cellular interactions and enable precision control of breathing, while providing only vague perceptions of organ states.


2020 ◽  
Vol 22 (Supplement_2) ◽  
pp. ii15-ii15
Author(s):  
Seth Malinowski ◽  
Mehdi Touat ◽  
Max Stockslager ◽  
Ross Giglio ◽  
Jack Geduldig ◽  
...  

Abstract Three-dimensional patient derived cultures hold great potential for use as personalized functional diagnostics, enabling more accurate preclinical evaluations of drug treatments compared to conventional cell lines. Optical imaging of live cells allows for continuous, time lapsed measurements, and can provide drug response data based on rich phenotypic changes of cell cultures. However, current imaging techniques based on 2D microscopy evaluation aren’t readily adaptable to evaluate the drug response of intact spheroids, which may better represent the in vivo environment and retain critical cellular interactions within the tumor microenvironment. Using the IncuCyte live cell imaging platform, we successfully imaged a large cohort (n = 77) of patient derived glioblastoma spheroid cultures and evaluated whether changes in sphere volume could be used as a direct measure of treatment response. Improving on the default Incucyte analysis software, we developed an R data processing pipeline better suited for spheroid measurements, which quantified the heterogeneity in GBM baseline spheroid growth, and calculated a drug response score based on spheroid changes in response to DNA damaging agents (TMZ as an example). Compared to conventional viability measurements, this novel 3D drug response score was found to accurately identify both drug sensitive and resistant spheroids and showed robust concordance with genomic biomarkers of response (NGS and MGMT promoter methylation) and patient outcomes. Additionally, we coupled the 3D drug score with known genetic data to explore other key pathways and genes involved in TMZ response. We provide here novel analysis methods and public code (Github) to advance the use of IncuCyte spheroid measurements, and deconvolute 3D spheroid drug response into a quantifiable statistic. These methods are adaptable to freshly isolated patient cells for rapid evaluation of treatment response in GBM patients while remaining widely applicable to other cancers such as pancreatic, colon, and non-cancer organoids/spheroids with 3D growth.


Glycobiology ◽  
2020 ◽  
Vol 30 (8) ◽  
pp. 490-499
Author(s):  
Marco D’Addio ◽  
Jasmin Frey ◽  
Vivianne I Otto

Abstract Vascular endothelia are covered with a dense glycocalix that is heavily sialylated. Sialylation of vascular glycoconjugates is involved in the regulation of cell–cell interactions, be it among endothelial cells at cell junctions or between endothelial and blood-borne cells. It also plays important roles in modulating the binding of soluble ligands and the signaling by vascular receptors. Here, we provide an overview over the sialylation-function relationships of glycoproteins expressed in the blood and lymphatic vasculature. We first describe cellular interactions in which sialic acid contributes in a stereospecific manner to glycan epitopes recognized by glycan-binding proteins. Our major focus is however on the rarely discussed examples of vascular glycoproteins whose biological functions are modulated by sialylation through other mechanisms.


2008 ◽  
Vol 14 (2) ◽  
pp. 117-125 ◽  
Author(s):  
Troy A. Baudino ◽  
Alex McFadden ◽  
Charity Fix ◽  
Joshua Hastings ◽  
Robert Price ◽  
...  

Patterning of cells is critical to the formation and function of the normal organ, and it appears to be dependent upon internal and external signals. Additionally, the formation of most tissues requires the interaction of several cell types. Indeed, both extracellular matrix (ECM) components and cellular components are necessary for three-dimensional (3-D) tissue formationin vitro. Using 3-D cultures we demonstrate that ECM arranged in an aligned fashion is necessary for the rod-shaped phenotype of the myocyte, and once this pattern is established, the myocytes were responsible for the alignment of any subsequent cell layers. This is analogous to thein vivopattern that is observed, where there appears to be minimal ECM signaling, rather formation of multicellular patterns is dependent upon cell–cell interactions. Our 3-D culture of myocytes and fibroblasts is significant in that it modelsin vivoorganization of cardiac tissue and can be used to investigate interactions between fibroblasts and myocytes. Furthermore, we used rotational cultures to examine cellular interactions. Using these systems, we demonstrate that specific connexins and cadherins are critical for cell–cell interactions. The data presented here document the feasibility of using these systems to investigate cellular interactions during normal growth and injury.


2015 ◽  
Vol 89 (19) ◽  
pp. 9706-9708 ◽  
Author(s):  
Lindsey M. Costantini ◽  
Erik L. Snapp

Many longstanding questions about dynamics of virus-cell interactions can be answered by combining fluorescence imaging techniques with fluorescent protein (FP) tagging strategies. Successfully creating a FP fusion with a cellular or viral protein of interest first requires selecting the appropriate FP. However, while viral architecture and cellular localization often dictate the suitability of a FP, a FP's chemical and physical properties must also be considered. Here, we discuss the challenges of and offer suggestions for identifying the optimal FPs for studying the cell biology of viruses.


2019 ◽  
Author(s):  
Alessandro Palma ◽  
Alberto Calderone ◽  
Andrea Cerquone Perpetuini ◽  
Federica Ferrentino ◽  
Claudia Fuoco ◽  
...  

AbstractMuscle regeneration is a complex process governed by the interplay between several muscle resident mononuclear cell populations. Following acute or chronic damage these cell populations are activated, communicate via cell-cell interactions and/or paracrine signals, influencing fate decisions via the activation or repression of internal signaling cascades. These are highly dynamic processes, occurring with distinct temporal and spatial kinetics. The main challenge toward a system level description of the muscle regeneration process is the integration of this plethora of inter- and intra-cellular interactions.We integrated the information on muscle regeneration in a web portal. The scientific content annotated in this portal is organized into two information layers representing relationships between different cell types and intracellular signaling-interactions, respectively. The annotation of the pathways governing the response of each cell type to a variety of stimuli/perturbations occurring during muscle regeneration takes advantage of the information stored in the SIGNOR database. Additional curation efforts have been carried out to increase the coverage of molecular interactions underlying muscle regeneration and to annotate cell-cell interactions.To facilitate the access to information on cell and molecular interactions in the context of muscle regeneration, we have developed Myo-REG, a web portal that captures and integrates published information on skeletal muscle regeneration.The muscle-centered resource we provide is one of a kind in the myology field. A friendly interface allows users to explore, approximately 100 cell interactions or to analyze intracellular pathways related to muscle regeneration. Finally, we discuss how data can be extracted from this portal to support in silico modeling experiments.


Sign in / Sign up

Export Citation Format

Share Document