2013-12-17 09:35:00 - High quality data generated for fast activating and desensitizing ligand gated ion channels
Rostock, Germany – Cytocentrics recently proved the rapid solution exchange capability of their fully automated patch clamp device CytoPatch™ 2 in GluA2 measurements. The unique CytoPatch™ chip technology and microfluidic features allow the ultrafast application and complete wash-out of agonists. This is a requirement for measuring fast activating and desensitizing ligand gated ion channels like GluA2.
GluA2 cDNA in CMV-based vector was a kind gift of Dr. Andrew Plested (FMP Berlin). Cells were transfected with standard transfection methods used in the Plested Lab. The results in the figure show whole-cell voltage-clamp recordings using HEK293 cells transiently transfected with cDNA, encoding the GluA2. Cells were clamped in the whole-cell configuration at a membrane potential of minus 70 mV. 10 mM Glutamate
was extracellularly applied to the cells three times for 10 milliseconds, leading to a fast activation of inward currents. The constant peak amplitudes confirm that the agonist is effectively removed in the intermediate wash-out phases. The insert shows an enlargement of the activation phase of the GluA2 mediated current. A rise time of 10 % to 90 % was achieved at 3.4 milliseconds, as displayed between the two vertical bars.
Cytocentrics Bioscience GmbH specializes in GLP-conform automated patch clamp devices, ready-to-use instant cells and ion channel screening services. Their CytoPatch™ 2 is a combination of hardware, software and proprietary microfluidic chip technology, generating data of the same high quality as manual patch clamp. Being a fully automated walk-away system, it can be operated by technical staff, saving time and resources. The device uses a 2K microfluidic chip, incorporating two real, independent patch pipettes within a small perfusion chamber. Due to its unique continuous perfusion system with fast wash-in/out capability, the CytoPatch™ 2 offers the perfect patch clamp platform for the study of both ligand- and voltage-gated ion channels in cell lines, stem cells and primary cell preparations.