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Vision Science Seminar

 

UPenn - Vision Research Center Confocal and Two-photon Imaging and Physiological Rig

This is a unique set up that is equipped for confocal imaging, two photon imaging, and physiological recordings (extracellular as well as intracellular and whole cell recordings; more information can be found below.

 

Location:

 

124 Anat-Chem Bldg

New users, contact:

Diego Contreras 121 Anat-Chem diegoc@mail.med.upenn.edu 215 898 8780
Sergei Nikonov 117 Anat-Chem sergein@mail.med.upenn.edu 215 898 7536

 

The sign-up using the VRC Rig Room Booking System. Alternatively, contact Marie Fina
Marie Fina 116 Anat-Chem mfina@retina.anatomy.upenn.edu 215 898 7536

 

Olympus FV1000 Confocal Microscope

Confocal Mode

  • Olympus BX61 upright with motorized stage.
  • Transmitted light, DIC and epifluorescence (DAPI, FITC, TRITC filters)
  • Simultaneous laser stimulation and imaging via SIM (SIMultaneous) Scanner
  • Fast scanning at 2 microseconds/pixel (16 fps at 256x256, bi-directional scanning), highly customizable scan sequences with a time controller
  • Other features: High-speed spectroscopy, FRET, FRAP, uncaging
  • Objectives: 10x dry and water, 40x oil and water, 60x oil and water.
  • 3 Laser units  with filter configuration providing the following:

 

Excitation wavelengths (nm)                            Emission Filters
405 425-475
488 500-550
515 525-565
559 575-625
635 650 Long Pass

Detectors

  • 3- Dedicated PMT for laser lines
  • diffraction gratings allow tunable wavelength selection for PMT1 and PMT2 and spectral scanning (lambda  scan, up to 100 nm/ms, up to 1nm resolution)
  • 1-DIC PMT (transmitted light detector)
  • A high resolution high sensitivity Infra red camera (18 MP still images, full HD at 30 fps video) to allow visual observation of the sample and electrode under IR light.
  •  

Two-Photon Mode

  • Ti-sapphire pulsed laser with pre-chirping tunable from 690 to 1020 nm
  • PMT for a green channel

Equipment for physiological recordings

  • Siskiyou MX7600 motorized 4-axis micromanipulator and a rotary motion controller.
  • Warner Instruments PC505B patch amplifier, and pClamp data acquisition and analysis software.
  • Perfusion chamber with associated equipment for oxygenating and heating. A two-channel, DC-powered, low-noise peristaltic pump for circulating solution.
  • Light stimulation system includes two computer controlled calibrated LEDs 528 nm or 475 nm with 500 nm and 455 nm interference filters (FWHM 10 nm). A full field light stimulus is directed to the sample via the condenser. The 455 blue LED allows simultaneous 2-photon imaging and light stimulation.
  • Dissection equipment including dissection microscope with IR illuminator and IR converters.
  • The microscope and related equipment are enclosed in a custom built Faraday cage providing light tight, low noise environment for the optical and electrophysiological recordings.