ephys rig

Welcome

Nearly 3 million Americans suffer from epilepsy. In one third of these patients available antiepileptic drugs or invasive surgical procedures are not effective. With an increased understanding of the molecular, electrophysiological and genetic bases of the epilepsies, hope for a cure emerges.  Understanding the fundamental basis of epilepsies, especially those that occur in children, and using this information to develop novel treatments is the mission of our laboratory.

Research in the BarabanLab is based on a multi-disciplinary strategy.  Techniques, briefly summarized here, include the use of in vitro brain slices for electrophysiological recording of membrane properties and synaptic function (using visualized patch-clamp and optogenetics methods); in vivo analysis of network function (using in silico probes and video-EEG monitoring); molecular analysis of gene expression using in situ hybridization and qPCR; immunohistochemical and morphological studies of neuronal structure and protein expression; pharmacological analysis including high-throughput drug screening in zebrafish; CRISPR-Cas9 based genome editing to generate epileptic zebrafish; and interneuron progenitor-based cell therapies

 

 

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Laboratory

In November 2014, the laboratory moved into completely remodeled laboratory space (HSE8; ~3000 sq. ft.) located on the main Parnassus Campus at UCSF.  Open lab space with 12 full-length benches is available to support “wet lab” functions and include benches dedicated to slice preparation, embryo dissection, Western blot, RT-PCR, cell injection, slice preparation, in situ hybridization and immunohistochemistry. A separate bench is designated for “RNase free” work.  A shared surgery and perfusion room with appropriate fume hood and sink is available as part of the 8th floor core facilities.  Four individual rooms, each with two complete electrophysiology set-ups equipped with IR-DIC rigs, are also available.  An additional dedicated electrophysiology plus confocal imaging rig is equipped with a 3i Intelligent Imaging Innovations imaging and acquisition system.  Each rig, in addition to appropriate amplifiers, micromanipulators, computer, slice heating and perfusion, LED light sources and filter wheels, and Olympus epifluoresence microscopes, is equipped with a vibration-free workstation, grounded/surge-protected electrical outlets and isolated air/vacuum lines.  One specialized set-up equipped with a Blackrock Microsystems integrated in vivo electrophysiology recording and behavioral imaging system. The laboratory also contains a dedicated zebrafish aquarium room with 5 stand-alone six-shelf housing systems and the capacity to hold over 10,000 adult zebrafish. Nikon Eclipse confocal microscope, Leica stereomicroscopes, gel boxes, centrifuges, Leica cryostat, hybridization oven, shakers and PCR (standard and qPCR) machines are available. Two DanioVision locomotion tracking systems running EthoVision software are also available.  

 

Approaches & Techniques
  • In vitro electrophysiology including visualized patch-clamp recording
  • In vivo electrophysiology and EEG in freely behaving animals
  • Optogenetic and chemogenetic approaches
  • Fast confocal GCaMP imaging in larval zebrafish
  • Rodent and zebrafish behavioral assays
  • Molecular biology (e.g., in situ hybridization and qPCR)
  • CRISPR/Cas9 genome editing
  • Immunohistochemistry
  • Confocal and light-sheet microscopy
  • Stereotactic surgery and cell transplantation

"You never change things by fighting the existing reality.  To change something, build a new model that makes the existing model obsolete" 

R. Buckminster Fuller