Methods and facilities

Intracellular setup

Our smaller EMI shielded room holds an electrophysiology setup for in vivo intracellular experiments. The setup is mainly used for recording from photoreceptors and lamina monopolar cells of various insects (Drosophila, Calliphora, Periplaneta, Apis, Bompus, etc.).

Stimulus hardware:

  • 15 LED revolver with neutral density filters + liquid light guide
  • computer-controlled LED driver electronics

Mechanical hardware:

  • Sand-filled, vibration free table
  • Cardan arm system for holding the specimen and positioning the light stimulus
  • Piezo-motor micromanipulator for the recording electrode

Intracellular amplifier:

  • NPI SEC-05L

Data acquisition:

  • PC + software + National Instruments data acquisition hardware
  • Filters, oscilloscopes, etc.

Other important features:

  • Air conditioning!

Patch-clamp setup #1

Patch-clamp setup is designed for whole-cell and single-channel recordings in photoreceptor cells but can also be utilized on recordings e.g. in cultured cells. The setup enables light stimulation via an in-house made LED wheel system (LEDs ranging from 355 to 690 nm in wavelength), bath temperature control and versatile pharmacological manipulations (exchange of both pipette and bath solutions).

  • Axopatch-1D amplifier (Molecular Devices)
  • Axiovert 35M inverted microscope (Zeiss)
  • P-285 micromanipulator (Sutter Instruments)
  • RSC-200 rapid solution changer (Bio Logic)
  • PClamp9 data acquisition and analysis software (Molecular Devices)

Patch-clamp setup #2 (under construction)

  • Model 2400 amplifier (A-M Systems)
  • CK 2 inverted microscope (Olympus)
  • Micromanipulator (Sensapex)

New patch-clamp setup is going to be an advanced version of the first patch-clamp setup with a modified design of the recording chamber, solution exchange system and light stimulation. Most of the equipment will be designed and made in-house.

Cockroach arena setup

Arena is used for studying behaviour of walking insect in unrestricted environment. Various patterning can be used to cover transparent walls and ultra thin optical fibre attached to the insect to record its movements relative to the patterning. Movement can also be recorded with synchronized night vision camera. Arena is illuminated with IR leds which are invisible to the eye but visible to the camera and fibre amplifier. Visible illumination is implemented with green leds.

  • 1 m diameter plastic coated arena
  • Continuously variable 850 nm IR led illumination
  • Continuously variable 550 nm green led illumination
  • 70 µm NA 0.12 optical fiber (CeramOptec UV70/77P12)
  • Femtowatt photoreceiver (ThorLabs PDF10A/M)
  • Night vision web camera (Microsoft, IR filter removed)
  • Custom made voltage and video recording software

Motionlab setup

This setup allows the study of various neuronal populations using both intra- and extracellular recording techniques. Research topics include motion vision, processing of visual signals by peripheral circuits and low-light vision. Research is facilitated by a high intensity video projector which permits the use of complex visual stimuli at various mean light intensity levels.

  • Mirage 6000 video projector (100Hz refresh rate)
  • Grayscale filters up to -6 decade
  • NPI SEC-10LX intracellular amplifier
  • A-M systems model 3600 16-channel amplifier

Virtual world setup

Virtual world setup is designed for large field stimulation for behavior and electrophysiological studies using DLP video projector. Air cushioned track ball is used as a measurement device in the behavior studies of walking insects. Both intracellular and extracellular measurement equipment are installed in the setup.

  • 40 cm diameter projection ball coated with barium sulfate
  • Custom fish eye lens system with slots for filters
    - Image covers 65 % of ball field
  • Commercial DLP video projector
    - Grey scale images
    - 480 Hz refresh rate with one computer controlled segment
    - Controlled by custom 3d software based on Ogre3d graphic engine
  • Air cushioned track ball
    - 10 cm diameter
    - 5 g weight
    - open and closed loop control
  • Adjustable heater provides ambient temperature control in a range of 20-40°C
  • Olfactory stimulator
    - Six channels
    - Constant flow
  • Intracellular equipment
  • Multichannel extracellular equipment

 

Last updated: 4.9.2013