Live‐cell imaging of cell signaling using genetically encoded fluorescent reporters

Ni, Qing Zhe; Mehta, Sohum; Zhang, Jin

doi:10.1111/febs.14134

Cited by 68 publications

(51 citation statements)

References 134 publications

(162 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Label‐free imaging of live cells has been demonstrated using interferometric scattering microscopy, showing intracellular organelles as well as topological characteristics of the membrane. Apart from new microscopy techniques, other live cell imaging techniques include new fluorescent probes as well as improvements in reporters for visualization of cell signalling dynamics …”

Section: Discussionmentioning

confidence: 99%

Cellular plasticity: A mechanism for homeostasis in the kidney

et al. 2020

View full text Add to dashboard Cite

Cellular plasticity is a topical subject with interest spanning a wide range of fields from developmental biology to regenerative medicine. Even the nomenclature is a subject of debate, and the underlying mechanisms are still under investigation. On top of injury repair, cell plasticity is a constant physiological process in adult organisms and tissues, in response to homeostatic challenges. In this review we discuss two examples of plasticity for the maintenance of homeostasis in the renal systemnamely the renin-producing juxtaglomerular cells (JG cells) and cortical collecting duct (CCD) cells. JG cells show plasticity through recruitment mechanisms, answering the demand for an increase in renin production. In the CCD, cells appear to have the ability to transdifferentiate between principal and intercalated cells to help maintain the highly regulated solute transport levels of that segment. These two cases highlight the complexity of plasticity processes and the role they can play in the kidney. K E Y W O R D Scollecting duct, JG cells, kidney, plasticity, renin 2 of 12 | ASSMUS et Al.

show abstract

Section: Discussionmentioning

confidence: 99%

Cellular plasticity: A mechanism for homeostasis in the kidney

et al. 2020

View full text Add to dashboard Cite

show abstract

“…Genetically encoded fluorescent reporters provide a powerful tool for understanding and quantifying dynamic signaling responses 35 . Various types of signaling reporters have been developed, including FRET sensors 36,37 , synthetic translocation reporters 38 , and native proteins that translocate between the nucleus and cytoplasm (FoxO1 10, 39 , p53 40 , SMAD2 41 , NFkB 42 , Erk 43 ).…”

Section: Reporter Optimization For Live Cell Imaging Studiesmentioning

confidence: 99%

High resolution AKT signaling in individual cells

Gross

Bucher

et al. 2018

Preprint

View full text Add to dashboard Cite

Cells sense and respond to their environment by activating distinct intracellular signaling pathways, however an individual cell's ability to faithfully transmit and discriminate environmental signals is thought to be limited. To assess the fidelity of signal transmission in the PI3K-AKT signaling pathway, we first developed an optimized genetically encoded sensor that had an increased dynamic range and reduced variation under basal conditions. We then used this reporter to track responses to varying doses of IGF-I in live cells and found that signaling responses from individual cells overlapped across a wide range of IGF-I doses, suggesting limited transmission accuracy. However, further analysis of individual cell traces revealed that responses were constant over time without stochastic fluctuations. We devised a new information theoretic approach to calculate the channel capacity using variance of the single cell time course data--rather than population-level variance as has been previously used-and predicted that cells were capable of discriminating multiple growth factor doses. We validated these predictions by tracking individual cell responses to multiple IGF-I doses and found that cells can accurately distinguish at least four different IGF-I concentrations, as demonstrated by their distinct responses. Furthermore, we found a similar discriminatory ability to pathway inhibition, as assessed by responses to the PI3K inhibitor alpelisib. Our studies indicate that cells can faithfully transmit an IGF-I input into a down-stream signaling response and that heterogeneous responses result from variation in the input-output relation across the population.These observations reveal the importance of viewing each cell as having its own communication channel and underscore the importance of understanding responses at the single cell level.

show abstract

“…Advances in imaging technologies -both in microscopes 11 and fluorescent reporters 12 -have significantly reduced the difficulty of acquiring live-cell imaging data while at the same time increasing the throughput and the number of systems amenable to this approach. Increasingly, image analysis is a bottleneck for discovery as there is a gap between our ability to collect and analyze data.…”

Section: Introductionmentioning

confidence: 99%

Caliban: Accurate cell tracking and lineage construction in live-cell imaging experiments with deep learning

Moen

Borba

Miller

et al. 2019

Preprint

View full text Add to dashboard Cite

Live-cell imaging experiments have opened an exciting window into the behavior of living systems. While these experiments can produce rich data, the computational analysis of these datasets is challenging. Single-cell analysis requires that cells be accurately identified in each image and subsequently tracked over time. Increasingly, deep learning is being used to interpret microscopy image with single cell resolution. In this work, we apply deep learning to the problem of tracking single cells in live-cell imaging data. Using crowdsourcing and a human-in-the-loop approach to data annotation, we constructed a dataset of over 11,000 trajectories of cell nuclei that includes lineage information. Using this dataset, we successfully trained a deep learning model to perform cell tracking within a linear programming framework. Benchmarking tests demonstrate that our method achieves state-of-the-art performance on the task of cell tracking with respect to multiple accuracy metrics. Further, we show that our deep learning-based method generalizes to perform cell tracking for both fluorescent and brightfield images of the cell cytoplasm, despite having never been trained on those data types. This enables analysis of live-cell imaging data collected across imaging modalities. A persistent cloud deployment of our cell tracker is available at http://www.deepcell.org.

show abstract

Live‐cell imaging of cell signaling using genetically encoded fluorescent reporters

Cited by 68 publications

References 134 publications

Cellular plasticity: A mechanism for homeostasis in the kidney

Cellular plasticity: A mechanism for homeostasis in the kidney

High resolution AKT signaling in individual cells

Caliban: Accurate cell tracking and lineage construction in live-cell imaging experiments with deep learning

Contact Info

Product

Resources

About