A multifunctional tool in the fight against stroke

Hemoproteins

The study is based on the discovery of a hemoprotein, Neuroglobin (Ngb), found in various parts of the brain, the pancreas and in the retina. Studies have shown that the protein has a protective functional role against hypoxia-related brain-damage. How this happens, is in the current situation, not clear.
In our group, techniques are developed and combined to explore the real-time response of functional single cells in different chemical environments. We aim to study and understand Ngbs effect on neurons signaling ability and electrophysiological activity under hypoxic (low oxygen) and anoxic (oxygen free) conditions using patch-clamp technique and optical spectroscopy.

Precise electrophysiological measurements

In order to perform precise electrophysiological measurements under well-controlled physiological conditions, patch clamp is included in a multi-functional system of a developed gas-tight microfluidic combined with optical manipulation technique and optical spectroscopy.
The setup is built on a vibration-free optical table (Technical Manufacturing Corporation, TMC, USA), where the the techniques are integrated on an inverted microscope (IX 71, Olympus, Japan).

• Patch clamp
  Patch Clamp - a thin glass pipette containing an electrode is attached to the cell membrane, a reference electrode in the surrounding fluid. The electrical signals of the cell is measured by the electrodes and analyzed by a patch clamp setup of headstage, signal amplifier (Heka EPC 7 Patch Clamp Amplifier, Germany) and software program.
• Microfluidic
  A micro-channel system is molded or etched into the transparent material, enables the electrophysiological measurements of single nerve cell in optimal control of rapid environmental changes.
• Optical Tweezers
  Light from NIR diode laser (IQ1A, Power Technology, USA) is used to trap and steer a single nerve cell through the micro-channel system towards the patch-clmap pipette.
• Optical Spectroscopy
  UV-Vis optical spectrometer (Ocean Optics, HR4000, USA) is used to monitor the cell's biochemical composition and oxygen binding by measuring the light absorption of substances at different wavelengths.

The project's long-term goals

The long-term goal is to develop a multifunctional system to understand the behavior, response and defense mechanisms of the cell in oxygen- shortage conditions related to Ngb, which can create new ways to interpret other hypoxia-related diseases such as Alzheimer's and Parkinson's.

• Patch clamp
  Patch Clamp - a thin glass pipette containing an electrode is applied to the cell membrane, a reference electrode in the surrounding fluid. The electrodes measure the membrane electrical signals that are recorded and analyzed through an amplifier (Heka EPC 7 Patch Clamp Amplifier, Germany).

• Microfluidic
  A micro duct molded or etched into the transparent material allows electrophysiological measurements of single nerve cell in the optimal control of rapid environmental changes.

• Optical Tweezers
  Light from the NIR diode laser (IQ1A, Power Technology, USA) is used to capture and control a nerve cell through micro-channel system in the patch-pipette clmap.
• Optical Spectroscopy
  UV-Vis optical spectrometer (Ocean Optics, HR4000, USA) is used to monitor the cell's biochemical composition and oxygen binding by measuring the light absorption of substances at different wavelengths.

The project's long-term goals

The project's long-term goal is to use this system to understand cell behavior, response and defense mechanisms in oxygen-poor conditions related to Ngb, which can create new ways to interpret other hypoxia-related diseases such as Alzheimer's and Parkinson's.