ArticlePloS one2013
A neuron-based screening platform for optimizing genetically-encoded calcium indicators.
Article in PloS one, 2013. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 39 papers.
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Who cites it
39 citing papers in PubMed, 88 citations in OpenAlex.
- A sensitive orange fluorescent calcium ion indicator for imaging neural activity.Nature communications · 2026Article
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- Glutamate indicators with increased sensitivity and tailored deactivation rates.Nature methods · 2026Article
- CaBLAM: a high-contrast bioluminescent CaNature methods · 2026Article
- Molecular recording of cellular protein kinase activity with chemical labeling.Nature chemical biology · 2025Article
- CaBLAM! A high-contrast bioluminescent CabioRxiv : the preprint server for biology · 2025Article
- A modular chemigenetic calcium indicator for multiplexed in vivo functional imaging.Nature methods · 2024Article
- Machine learning-guided engineering of genetically encoded fluorescent calcium indicators.Nature computational science · 2024Article
- Neurophotonic methods in approach to in vivo animal epileptic models: Advantages and limitations.Epilepsia · 2024Review
- Optogenetic Microwell Array Screening System: A High-Throughput Engineering Platform for Genetically Encoded Fluorescent Indicators.ACS sensors · 2023Article
- A general method for the development of multicolor biosensors with large dynamic ranges.Nature chemical biology · 2023Article
- Functional imaging-guided cell selection for evolving genetically encoded fluorescent indicators.Cell reports methods · 2023Article
- Glutamate indicators with improved activation kinetics and localization for imaging synaptic transmission.Nature methods · 2023Article
- Article
- Article
- Fluorescent and Bioluminescent Calcium Indicators with Tuneable Colors and Affinities.Journal of the American Chemical Society · 2022Article
- Design and Initial Characterization of a Small Near-Infrared Fluorescent Calcium Indicator.Frontiers in cell and developmental biology · 2022Article
- Designs, applications, and limitations of genetically encoded fluorescent sensors to explore plant biology.Plant physiology · 2021Review
- High-throughput Analysis of Synaptic Activity in Electrically Stimulated Neuronal Cultures.Neuroinformatics · 2021Article
- The HaloTag as a general scaffold for far-red tunable chemigenetic indicators.Nature chemical biology · 2021Article
Corrections and comments
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Authors and funding
15 authors at 2 institutions in 1 country.
Funding
Abstract
Fluorescent protein-based sensors for detecting neuronal activity have been developed largely based on non-neuronal screening systems. However, the dynamics of neuronal state variables (e.g., voltage, calcium, etc.) are typically very rapid compared to those of non-excitable cells. We developed an electrical stimulation and fluorescence imaging platform based on dissociated rat primary neuronal cultures. We describe its use in testing genetically-encoded calcium indicators (GECIs). Efficient neuronal GECI expression was achieved using lentiviruses containing a neuronal-selective gene promoter. Action potentials (APs) and thus neuronal calcium levels were quantitatively controlled by electrical field stimulation, and fluorescence images were recorded. Images were segmented to extract fluorescence signals corresponding to individual GECI-expressing neurons, which improved sensitivity over full-field measurements. We demonstrate the superiority of screening GECIs in neurons compared with solution measurements. Neuronal screening was useful for efficient identification of variants with both improved response kinetics and high signal amplitudes. This platform can be used to screen many types of sensors with cellular resolution under realistic conditions where neuronal state variables are in relevant ranges with respect to timing and amplitude.
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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.