Microelectrode Array (MEA) Manual Information in this document is subject to change without notice. No part of this document may be reproduced or transmitted without the express written permission of Multi Channel Systems MCS GmbH. While every precaution has been taken in the preparation of this document, the publisher and the author assume no responsibility for errors or omissions, or for damages resulting from the use of information contained in this document or from the use of programs and source code that may accompany it. In no event shall the publisher and the author be liable for any loss of profit or any other commercial damage caused or alleged to have been caused directly or indirectly by this document. © 2010 Multi Channel Systems MCS GmbH. All rights reserved. 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Table of Contents 1 Introduction 7 1.1 About this Manual 7 2 Important Information and Instructions 9 2.1 Operator's Obligations 9 2.2 Guarantee and Liability 9 2.3 Important Safety Advice 10 3 Microelectrode Arrays (MEAs) — Overview 11 3.1 Extracellular Recording with Microelectrode Arrays 11 3.2 MEA Design and Production 12 3.3 Electrodes, Tracks, and Insulation 13 4 MEA Types and Layouts 14 4.1 Standard Electrode Numbering 15 4.2 Standard MEA 16 4.3 HighDense MEA 17 4.4 HexaMEA 18 4.5 ThinMEA 19 4.6 3-D MEA 20 4.7 EcoMEA 21 4.8 Stimulation MEA 22 4.9 Perforated MEA 23 4.10 6-Well MEA 24 4.11 256MEA 25 4.12 4QMEA1000 26 4.13 FlexMEA 27 4.14 EcoFlexMEA 29 4.15 MEA Signal Generator 31 5 MEA Handling 32 5.1 Hydrophilic Surface Treatment 32 5.1.1 Plasma Cleaning 32 5.1.2 Protein Coating 32 5.1.3 Preculturing 33 5.2 Sterilization 33 5.2.1 Sterilization with Ethanol and UV Light 33 5.2.2 Steam Sterilization (Autoclavation) 33 5.2.3 Dry Heat Sterilization 33 5.2.4 Sterilization with Hot Water 33 MEA Manual 5.3 MEA Storage 34 5.4 MEA Coating 34 5.4.1 Coating with Nitrocellulose 34 5.4.2 Coating with Polyethyleneimine (PEI) plus Laminin36 5.4.3 Coating with Polyornithine (plus Laminin) 38 5.4.4 Coating with Poly-D-Lysine (plus Laminin) 39 5.4.5 Coating with Poly-D-Lysine (plus Fibronectin) 40 5.4.6 Coating with Fibronectin 41 5.4.8 Coating with Collagen 42 5.5 Cleaning of used MEAs 43 5.5.1 General Recommendations for Cleaning MEAs 43 5.5.2 Cleaning of 3-D MEAs 43 5.5.3 Cleaning of pMEAs 43 5.5.4 Cleaning of EcoMEAs 43 5.5.5 Cleaning of EcoFlexMEAs 44 5.5.6 Cleaning of FlexMEAs 44 5.5.7 Removing Nitrocellulose Coating 44 5.5.8 MEA Cleaning with EDTA-Collagenase 44 5.5.9 MEA Cleaning with Terg-A-Zyme 45 6 Culture Chamber Options 46 6.1 Sealed MEA Culture Dish 46 6.2 MEA Culture Chamber with Lid 47 6.3 Removable Recording Chamber 47 7 Recording with MEAs 49 7.1 Mounting the MEA 49 7.1.1 Cleaning the Contact Pads 49 7.1.2 Positioning the MEA 49 7.1.3 Grounding the Bath 49 7.2 General Performance / Noise Level 50 8 Stimulation 53 8.1 Using MEA Electrodes for Stimulation 53 8.2 Capacitive Behavior of Stimulating Electrodes 54 8.3 Aspects of Electrode Size and Material 55 8.4 Recommended Stimulus Amplitudes and Durations 56 9 Troubleshooting 57 9.1 About Troubleshooting 57 9.2 Technical Support 57 9.3 Noise on Single Electrodes 58 9.4 Overall Noise / Unsteady Baseline 60 9.5 Missing Spikes or Strange Signal Behavior 61 4 10 Appendix 63 10.1 Contact Information 63 10.2 Ordering Information 64 10.2.1 MEA-Systems 64 10.2.2 USB-MEA-Systems 65 10.2.3 Stimulus Generators 67 10.2.4 ME-Systems 68 10.2.5 ME Amplifiers 69 10.2.6 MEA Amplifiers 69 10.2.7 Accessories 70 10.3 Safe Charge Injection Limits 71 10.4 MEA Data Sheet 72 Introduction 1 Introduction 1.1 About this Manual The MEA Manual comprises all important information about the microelectrode arrays (MEA) for use with (USB-) MEA- or ME-Systems from Multi Channel Systems. The MEA Manual focuses on general information on the MEA design, use, and handling, and more specific information on different MEA types. It also includes recommendations on sterilization, coating, and cleaning procedures, from scientifical papers or from recommendations of other MEA users. For more details on issues that refer to the amplifier, like grounding or mounting the MEA, please refer to the manual for the MEA amplifier you use. You will find more information about the MEA-System and its components in general, especially the data acquisition card, in the MEA-System Manual. For more details on the data acquisition and analysis program MC_Rack, please refer to the MC_Rack Manual. It is assumed that you have already a basic understanding of technical terms. No special skills are required to read this manual. The components and also the manual are part of an ongoing developmental process. Please understand that the provided documentation is not always up to date. Please check the MCS Web site (www.multichannelsystems.com) from time to time for downloading up-to-date manuals. Those parts in this manual that refer to the applications, and not to the product itself, for example, coating of MEAs, are only a summary of published information from other sources (see references) and has the intention of helping users finding the appropriate information for setting up their experiments. Multi Channel Systems MCS GmbH has not tested or verified this information. Multi Channel Systems MCS GmbH does not guarantee that the information is correct. Multi Channel Systems MCS GmbH recommends to refer to the referenced literature for planning and executing any experiments. 7 Important Information and Instructions 2 Important Information and Instructions 2.1 Operator's Obligations The operator is obliged to allow only persons to work on the device, who . are familiar with the safety at work and accident prevention regulations and have been instructed how to use the device; . are professionally qualified or have specialist knowledge and training and have received instruction in the use of the device; . have read and understood the chapter on safety and the warning instructions in this manual and confirmed this with their signature. It must be monitored at regular intervals that the operating personnel are working safely. Personnel still undergoing training may only work on the device under the supervision of an experienced person. 2.2 Guarantee and Liability The General conditions of sale and delivery of Multi Channel Systems MCS GmbH always apply. The operator will receive these no later than on conclusion of the contract. Multi Channel Systems MCS GmbH makes no Guarantee as to the accuracy of any and all tests and data generated by the use of the device or the software. It is up to the user to use good laboratory practice to establish the validity of his findings. Guarantee and liability claims in the event of injury or material damage are excluded when they are the result of one of the following. Improper use of the device . Improper installation, commissioning, operation or maintenance of the device . Operating the device when the safety and protective devices are defective and/or inoperable . Non-observance of the instructions in the manual with regard to transport, storage, installation, commissioning, operation or maintenance of the device . Unauthorized structural alterations to the device . Unauthorized modifications to the system settings . Inadequate monitoring of device components subject to wear . Improperly executed and unauthorized repairs . Unauthorized opening of the device or its components . Catastrophic events due to the effect of foreign bodies or acts of God 9 MEA Manual 2.3 Important Safety Advice Warning: Make sure to read the following advice prior to install or to use the device and the software. If you do not fulfill all requirements stated below, this may lead to malfunctions or breakage of connected hardware, or even fatal injuries. Warning: Obey always the rules of local regulations and laws. Only qualified personnel should be allowed to perform laboratory work. Work according to good laboratory practice to obtain best results and to minimize risks. The product has been built to the state of the art and in accordance with recognized safety engineering rules. The device may only . be used for its intended purpose; . be used when in a perfect condition. Improper use could lead to serious, even fatal injuries to the user or third parties and damage to the device itself or other material damage. Warning: The device and the software are not intended for medical uses and must not be used on humans. Malfunctions which could impair safety should be rectified immediately. Regard the technical specifications of the various MEA types, especially the temperature range and the safe charge injection limits for stimulation. Do not autoclave or expose 3-D MEAs, pMEAs or FlexMEAs to heat.