Sensitive red protein calcium indicators for imaging neural activity

Dana, Hod; Mohar, Boaz; Sun, Yi; Narayan, Sujatha; Gordus, Andrew; Hasseman, J.; Tsegaye, Getahun; Holt, Graham T.; Hu, Amy; Walpita, Deepika; Patel, Ronak; Macklin, John J.; Bargmann, Cornelia I.; Ahrens, Misha B.; Schreiter, Eric R.; Jayaraman, Vivek; Looger, Loren L.; Svoboda, Karel; Kim, Douglas S.

doi:10.7554/elife.12727

Cited by 877 publications

(855 citation statements)

References 52 publications

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“…Simultaneous efforts have focused on the development of compounds with enhanced multiphoton properties [51]. In addition, improved red-shifted calcium indicators with advantages for deep in vivo imaging are being designed to rival the sensitivity of GCaMP6 [26]. Together, the characterization of existing contrast agents and the development of novel chromophores for MPM will identify and produce probes that are sufficiently bright and photostable to overcome background fluorescence and scattering in vivo .…”

Section: Availability Of Fluorophoresmentioning

confidence: 99%

Deep tissue imaging with multiphoton fluorescence microscopy

Miller

Jarrett

Hassan

et al. 2017

Current Opinion in Biomedical Engineering

118

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We present a review of imaging deep-tissue structures with multiphoton microscopy. We examine the effects of light scattering and absorption due to the optical properties of biological sample and identify 1,300 nm and 1,700 nm as ideal excitation wavelengths. We summarize the availability of fluorophores for multiphoton microscopy as well as ultrafast laser sources to excite available fluorophores. Lastly, we discuss the applications of multiphoton microscopy for neuroscience.

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Section: Availability Of Fluorophoresmentioning

confidence: 99%

Deep tissue imaging with multiphoton fluorescence microscopy

Miller

Jarrett

Hassan

et al. 2017

Current Opinion in Biomedical Engineering

118

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show abstract

“…Localization sequences further restrict gene expression to distinct cellular compartments such as the nucleus (via one or more nuclear localization signals (NLS)) (Fig. 2), cytosol (via a nuclear exclusion signal (NES) 58 ), or cell membrane (via farnesylation 55 , the CD4-2 59 transmembrane (TM) targeting domain, or PDZ 60 protein-protein interaction domains) (Fig. 3d).…”

Section: Refer Tomentioning

confidence: 99%

Widespread and targeted gene expression by systemic AAV vectors: Production, purification, and administration

Challis

Kumar

Chan

et al. 2018

Preprint

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We recently developed novel AAV capsids for efficient and noninvasive gene transfer across the central and peripheral nervous systems. In this protocol, we describe how to produce and systemically administer AAV-PHP viruses to label and/or genetically manipulate cells in the mouse nervous system and organs including the heart. The procedure comprises three separate stages: AAV production, intravenous delivery, and evaluation of transgene expression.The protocol spans eight days, excluding the time required to assess gene expression, and can be readily adopted by laboratories with standard molecular and cell culture capabilities. We provide guidelines for experimental design and choosing the capsid, cargo, and viral dose appropriate for the experimental aims. The procedures outlined here are adaptable to diverse biomedical applications, from anatomical and functional mapping to gene expression, silencing, and editing.

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“…The activity of neurons can be assessed through the use of different genetically encoded indicators of neural activity (Grienberger and Konnerth 2012;Masuyama et al 2012;Broussard et al 2014;Fosque et al 2015;Gao et al 2015;Dana et al 2016). For example, genetically encoded calcium indicators (GECIs) increase their fluorescence as calcium levels rise in active neurons.…”

Section: Assessing Functional Connectivity Among Neuronsmentioning

confidence: 99%

Targeted manipulation of neuronal activity in behaving adult flies

Hampel

Seeds

2016

Preprint

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The ability to control the activity of specific neurons in freely behaving animals provides an effective way to probe the contributions of neural circuits to behavior. Wide interest in studying principles of neural circuit function using the fruit fly Drosophila melanogaster has fueled the construction of an extensive transgenic toolkit for performing such neural manipulations. Here we describe approaches for using these tools to manipulate the activity of specific neurons and assess how those manipulations impact the behavior of flies. We also describe methods for examining connectivity among multiple neurons that together form a neural circuit controlling a specific behavior. This work provides a resource ABSTRACTThe ability to control the activity of specific neurons in freely behaving animals provides an effective way to probe the contributions of neural circuits to behavior. Wide interest in studying principles of neural circuit function using the fruit fly Drosophila melanogaster has fueled the construction of an extensive transgenic toolkit for performing such neural manipulations. Here we describe approaches for using these tools to manipulate the activity of specific neurons and assess how those manipulations impact the behavior of flies. We also describe methods for examining connectivity among multiple neurons that together form a neural circuit controlling a specific behavior. This work provides a resource for researchers interested in examining how neurons and neural circuits contribute to the rich repertoire of behaviors performed by flies.

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Sensitive red protein calcium indicators for imaging neural activity

Cited by 877 publications

References 52 publications

Deep tissue imaging with multiphoton fluorescence microscopy

Deep tissue imaging with multiphoton fluorescence microscopy

Widespread and targeted gene expression by systemic AAV vectors: Production, purification, and administration

Targeted manipulation of neuronal activity in behaving adult flies

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