A5054437859- Advanced Fluorescence Microscopy Techniques
- Photoreceptor and optogenetics research
- Retinal Development and Disorders
- Cell Image Analysis Techniques
- Neuroscience and Neural Engineering
- Neuroscience and Neuropharmacology Research
- Neural dynamics and brain function
- Neurogenesis and neuroplasticity mechanisms
- Neuroinflammation and Neurodegeneration Mechanisms
- Spectroscopy Techniques in Biomedical and Chemical Research
- Digital Holography and Microscopy
- bioluminescence and chemiluminescence research
- Neurobiology and Insect Physiology Research
- Advanced X-ray Imaging Techniques
- Optical Coherence Tomography Applications
- S100 Proteins and Annexins
- Anesthesia and Neurotoxicity Research
- Zebrafish Biomedical Research Applications
- Multiple Sclerosis Research Studies
- Traumatic Brain Injury and Neurovascular Disturbances
- Near-Field Optical Microscopy
- Optical Imaging and Spectroscopy Techniques
- Long-Term Effects of COVID-19
- Microwave Imaging and Scattering Analysis
- Traumatic Brain Injury Research
Cleveland Clinic Lerner College of Medicine
2018-2024
Cleveland Clinic
2018-2024
Case Western Reserve University
2019-2023
Cerner (United States)
2021
Janelia Research Campus
2014-2019
Howard Hughes Medical Institute
2014-2019
Technion – Israel Institute of Technology
2011-2015
Helix (United States)
2015
Genetically encoded calcium indicators (GECIs) allow measurement of activity in large populations neurons and small neuronal compartments, over times milliseconds to months. Although GFP-based GECIs are widely used for vivo neurophysiology, with red-shifted excitation emission spectra have advantages imaging because reduced scattering absorption tissue, a consequent reduction phototoxicity. However, current red inferior the state-of-the-art GCaMP6 detecting quantifying neural activity. Here...
Genetically-encoded calcium indicators (GECIs) facilitate imaging activity of genetically defined neuronal populations in vivo. The high intracellular GECI concentrations required for vivo are usually achieved by viral gene transfer using adeno-associated viruses. Transgenic expression GECIs promises important advantages, including homogeneous, repeatable, and stable without the need invasive virus injections. Here we present generation characterization transgenic mice expressing GCaMP6s or...
The identification of active neurons and circuits in vivo is a fundamental challenge understanding the neural basis behavior. Genetically encoded calcium (Ca(2+)) indicators (GECIs) enable quantitative monitoring cellular-resolution activity during However, such require online within limited field view. Alternatively, post hoc staining immediate early genes (IEGs) indicates highly cells entire brain, albeit with poor temporal resolution. We designed fluorescent sensor, CaMPARI, that combines...
Marking functionally distinct neuronal ensembles with high spatiotemporal resolution is a key challenge in systems neuroscience. We recently introduced CaMPARI, an engineered fluorescent protein whose green-to-red photoconversion depends on simultaneous light exposure and elevated calcium, which enabled marking active populations single-cell subsecond resolution. However, CaMPARI (CaMPARI1) has several drawbacks, including background low slow kinetics reduced fluorescence after chemical...
Genetically encoded calcium ion (Ca2+) indicators (GECIs) are indispensable tools for measuring Ca2+ dynamics and neuronal activities in vitro vivo. Red fluorescent protein (RFP)-based GECIs have inherent advantages relative to green protein-based due the longer wavelength light used excitation. Longer is associated with decreased phototoxicity deeper penetration through tissue. GECI can also enable multicolor visualization blue- or cyan-excitable fluorophores. Here we report development,...
Calcium imaging is commonly used to measure the neural activity of large groups neurons in mice. Genetically encoded calcium indicators (GECIs) can be delivered for this purpose using non-invasive genetic methods. Compared viral gene transfer, transgenic targeting GECIs provides stable long-term expression and obviates need invasive injections. Transgenic mice expressing green GECI GCaMP6 are already widely used. Here we present generation characterization sensitive red jRGECO1a, driven by...
Damage to the myelin sheath that protects axons in central nervous system is a hallmark pathology of demyelinating diseases like multiple sclerosis. Cuprizone-induced demyelination mice common model for studying and remyelination. However, relationship between damage recovery functional properties ensheathed neurons remains poorly understood. Using concurrent monitoring hippocampal myelination neuronal firing rates same mouse, we assessed longitudinal changes during cuprizone consumption...
Temporal focusing is a simple approach for achieving tight, optically sectioned excitation in nonlinear microscopy and multiphoton photo-manipulation. Key applications advantages of temporal involve propagation through scattering media, but the progressive broadening focus has not been characterized. By combining detailed geometrical optics model with Monte-Carlo simulations we introduce validate simulation strategy predicting characteristics non-scattering media. The width increasing depth...
Abstract Calcium imaging with genetically encoded calcium indicators (GECIs) is routinely used to measure neural activity in intact nervous systems. GECIs are frequently one of two different modes: track large populations neuronal cell bodies, or follow dynamics subcellular compartments such as axons, dendrites and individual synaptic compartments. Despite major advances, still limited by the biophysical properties existing GECIs, including affinity, signal-to-noise ratio, rise decay...
Abstract Current methods for recording large-scale neuronal activity from behaving mice at single-cell resolution require either fixing the mouse head under a microscope or attachment of device to animal’s skull. Both these options significantly affect animal behavior and hence also recorded brain patterns. Here, we introduce different method acquire snapshots cortical maps freely-moving using calcium sensor called CaMPARI. CaMPARI has unique property irreversibly changing its color green...
Line illumination geometries have advantageous properties for temporal focusing nonlinear microscopy. The characteristics of line (LITEF) in transparent and scattering media are studied here both experimentally using numerical model simulations. We introduce an approximate analytical formula the dependence axial sectioning on laser microscope's parameters. Furthermore, we show that LITEF is more robust to tissue than wide-field focusing, can penetrate much deeper into while maintaining good...
Calcium-modulated photoactivatable ratiometric integrator (CaMPARI) is a calcium ion (Ca 2+ )- and light-dependent genetically encoded fluorescent activity that can capture snapshots of neuronal through an irreversible process known as photoconversion. This unique property was previously used to label neurons based upon their tuning properties in order map synaptic connectivity record large-scale freely moving mice without attaching any mechanical device them. The latest version CaMPARI...
A simple technique for remote scanning of the focal plane in temporal focusing multiphoton microscopy is demonstrated both theoretically and experimentally. new on-axis light propagation optical setup design enables this scanning, which was considered not feasible previous studies. The axially displaced by movement a device, consisting double prism grating, optionally cylindrical lens. displacement linear, its slope inversely proportional to square system's magnification.
Abstract Genetically encoded calcium indicators (GECIs) allow measurement of activity in large populations neurons and small neuronal compartments, over times milliseconds to months. Although GFP-based GECIs are widely used for vivo neurophysiology, with red-shifted excitation emission spectra have advantages imaging because reduced scattering absorption tissue, a consequent reduction phototoxicity. However, current red inferior the state-of-the-art GCaMP6 detecting quantifying neural...
Our understanding of neural information processing could potentially be advanced by combining flexible three-dimensional (3-D) neuroimaging and stimulation. Recent developments in optogenetics suggest that neurophotonic approaches are principle highly suited for noncontact stimulation network activity patterns. In particular, two-photon holographic optical (2P-HONS) has emerged as a leading approach multisite 3-D excitation, it with temporal focusing (TF) further enables axially confined yet...
Demyelination of axons in the central nervous system (CNS) is a hallmark multiple sclerosis (MS) and other demyelinating diseases. Cycles demyelination, followed by remyelination, appear majority MS patients are associated with onset quiescence disease-related symptoms, respectively. Previous studies human animal models have shown that vast demyelination accompanied wide-scale changes to brain activity, but details this process poorly understood. We used electrophysiological recordings...
Traumatic brain injury (TBI)-induced axonal degeneration leads to acute and chronic neuropsychiatric impairment, neuronal death, accelerated neurodegenerative diseases of aging, including Alzheimer's Parkinson's diseases. In laboratory models, is traditionally studied through comprehensive postmortem histological evaluation integrity at multiple time points. This requires large numbers animals power for statistical significance. Here, we developed a method longitudinally monitor functional...