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Cardio 2D Manual - ALA Scientific Instruments

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1. 27 pp 27 Propagation time ssssssse 27 AMPUTIO ibi 27 Detection deadtime 27 HB detection iiie nt vvv ai 27 RISO TIMES ad 27 Eo AMA A ee 27 GENET I TEPORE 27 SUO labiale 27 N Noise level 45 R Re entry cycles 2 RR interval 2 36 62 70 S Single channel window 15 56 Software 9 InStalliTiO uta vd pe tane bb ao idu 9 Bier M 9 MC Card Data Acquisition 9 System requirements 9 USB ME System Data Acquisition 9 Stimulation 36 Inter Pulse Interval 36 Pulse Shape 36 BIDS Cuaderno iena 36 SVMMCUIC i usi Ae eae 36 Voltage PUS eE iii 36 Stimulus Generator 36 Cardio 2D Manual T Technical Support 86 81 Troubleshooting 87
2. gt 15 11 40 gt Recording 2 Test Color map gt 15 11 43 gt Experiment stop 00 00 00 000000 Ga ea Be u 15 12 40 gt Experiment stop Log file Single channel view Latency 500jV 10s 2 RefCh47 Ch48 Main Window Overview The main window of Cardio 2D consists of two window panes The left Control Window is for controlling and documentation of the experiment The bigger window on the right site the MEA Window displays heartbeats in different characteristics in real time monitoring You have one single Control Window only but depending on the connected hardware you can have up to four different MEA tabs in the MEA Window If you connect for example an USB ME256 System for data acquisition it is possible to connect up to four MEAs Each MEA tab will display the heartbeats with different settings in the five views of the MEA Window independent from each other Via Control Window you start and stop the experimental protocol simultaneously MEA 1 MEA 2 MEA 3 MEA 4 uw 15 Cardio 2D Manual Control Window The experimental protocol is visualized by three icons on the left orange control view The icons can be clicked for editing the protocol Below the control view the log file documents the ongoing experimental proceedings MEA Window The MEA Window is divided into five single views All of them display heartbeats in different characteristics Recorded data
3. 42 Cardio 2D Software Click the Record Movie icon The dialog Movie Recorder appears MEA 1 Movie Recorder a Movie Type MN Frame Rate 30 Frames s Compression On E File Mame Start Cancel The movie of the color map will be recorded in AVI format The AVI format is a common format You just need a Media Player to replay the video You can choose the Frame Rate between one and hundred frames per second Select Compression of the movie with On or deselect the compression with Off Save the movie by clicking the Save icon al and select a file name and the path to the folder you want to store it Recording the color map movie starts as soon as you click Start 43 5 1 Experimental Proceeding General Performance Noise Level Please use the provided model test probe to test the amplifier immediately after installation The test model probe is already mounted on the amplifier It simulates a MEA with a resistor of 220 kO and a 1 nF capacitor between bath and electrode Use the Cardio 2D program to record from the test model probe and to check the amplifier This also presents an opportunity for you to learn about the program before starting a real experiment Please keep the test model probe at a safe place in case that you will need it later for troubleshooting Starting the recording For testing the amplifier it is not necessary to run a complete experiment You may choose to
4. Arrows Dm hd asinum 20 ms Auto Size w x H 200 x 00 Interpolation Bicubic In this dialog you can define the settings for the color map 40 Cardio 2D Software Color Map The usual used color map for cardiac issues is the Jet Invers map but you can also choose Hot Cool or Grey maps To remove the colors from the map click Off The isochrones will remain only O AI o E Mea o Oo o Contour The drop down menu is for changing the color of the contour for the isochrones into Black Jet Hot Cool and Grey Or you can remove the lines with Off The time step is defined in the dialog Arrows The feature is not jet implemented in this version of Cardio 2D As soon as it will be implemented you find a Read Me on MCS web site and you can download the new version of Cardio 2D Minimum Maximum You have to define the minimum and the maximum of the scale of the color map That means you define the time frame in which the excitation pattern are displayed in the color map Usually you start with zero in the minimum The excitation pattern will run from the pace maker over the map to the maximum The widest selectable time frame for the color map spans from 0 us in the minimum to 10000 ms in the maximum Auto The Cardio 2D program automatically finds the minimum and maximum of each heart beat and adapts the scaling of the color map It is recommended to use the auto scaling function Size
5. Introduction About this Manual This manual comprises all important information about the first installation of the hardware and software and about the daily work with the instrument It is assumed that you have already a basic understanding of technical and software terms No special skills are required to read this manual If you are using the device for the first time please read the important safety advice before installing the hardware and software where you will find important information about the installation and first steps The printed manual and Help are basically the same so it is up to you which one you will use The Help offers you the advantage of scrolling through the text in a non linear fashion picking up all information you need especially if you use the Index the Search function and the Browse Sequences If you are going to read larger text passages however you may prefer the printed manual The device and the software are part of an ongoing developmental process Please understand that the provided documentation is not always up to date The latest information can be found in the Help Check also the MCS Web site www multichannelsystems com for downloading up to date manuals and Help files Welcome to Cardio 2D and Cardio 2D The Cardio 2D software consists of two main parts The recording part is called Cardio 2D program It is for controlling the amplifier and the stimulator and for scheduling and runni
6. In the settings file all information about the currently analyzed Cardio 2D result or raw data file is displayed in a tree view MEA Window The window on the right side is divided into four separate window panes and has two tabs Field Potential and Heartbeat The setting of the tabs influences each of the four separate windows Select Field Potential tab Please see the upper picture The single channel view on top of the right window panes shows field potentials Recorded data traces are displayed in the MEA electrodes pane If you select one or more than one MEA electrodes in this window pane they are displayed in a overlay plot in the overlay plots window below The color map displays the spreading of the excitation of a heartbeat from the pace maker or the earliest time point detected if the pace maker is not on the MEA to the outer regions of the cardiac cell culture or tissue The isochrones are indicated by black lines The colors indicates the time distance between the first signal and outer regions of excitation Red color represents the pace maker when using the color map jet inverse Below the map the actual time stamp is displayed The color map corresponds with the single channel view and with the field potential windows 52 Cardio 2D Analyzer Software Select Heartbeat tab Please see the picture below The single channel view on top of the right window panes shows amplitude and rate of the heartbeats of
7. multichannel systems Cardio 2D 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 Printed 27 01 2010 Multi Channel Systems MCS GmbH AspenhaustraBe 21 72770 Reutlingen Germany Fon 49 71 21 90 925 0 Fax 49 71 21 90 92 5 11 info multichannelsystems com www multichannelsystems com Microsoft and Windows are registered trademarks of Microsoft Corporation Products that are referred to in this document may be either trademarks and or registered trademarks of their respective holders and should be noted as such The publisher and the author make no claim to these trademark 1 1 1 2 1 3 1 4 2 1 2 2 2 3 2 4 2 5 3 1 3 2 3 3 4 1 4 2 4 3 5 1 5 2 5 3 5 4 Table of Contents Introduction About this M
8. Running a viability test Before you start recording a viability test is recommended The data is not recorded The viability test monitors whether the MEA electrodes are acceptable for recording and the cell culture or tissue is fine You can check the signal detection parameters and focus onto single channels Too rigid setting will lead to data loss that is the cardiac signal is not detected at all Too loose settings will lead to a wrong signal detection and thus to wrong results After adjusting all parameters in the different window panes you can start recording The log file documents the experimental proceeding 22 Cardio 2D Software Click Start to start recording The Start icon changes into Stop icon duration of recording is counted up in seconds Stop recording manual by clicking on Stop icon and the Cardio 2D generates three types of files the Raw Data file c2d the Result file c2r and the Settings file c2s Raw data file and the associated settings file always belong together in the result file the settings file is included The log file is not automatically generated but by users command Basic Click Basic The Basic Settings dialog appears Basic Settings J Switch Recording Timer Llickleon Phase Duration Timer Enter Comment C Recording Complete z The duration of a recording phase can be influenced in two ways It is possible to start a new recording pha
9. Warning The operating system settings of the data acquisition computer were settings can lead to program instabilities and data loss Data Acquisition with the external USB ME Systems The USB ME256 System is a stand alone solution for acquiring data from up to 256 channels from up to four MEAs It can replace the internal computer hardware MC Card The analog input signals are acquired and digitized by the USB ME256 System and the digital electrode signals are transmitted to the connected computer via universal serial bus High Speed USB 2 0 Thus it is possible to use any computer for data processing Recorded data is graphed analyzed and reviewed with Cardio 2D and Cardio 2D program The USB ME256 System supports 256 channels with an amplification factor 1 that means no amplification data acquisition only You can connect for example up to four MEA1060 amplifiers via MCS 68 pin high grade cables The USB ME128 System and the USB ME64 System are in principle the same devices as the USB ME256 System but with 128 or 64 electrode channels respectively Data Acquisition with the external USB MEA256 Systems The USB MEA256 System is a compact and portable stand alone solution for MEA recordings with integrated amplification data acquisition and analog digital conversion The system acquires data from up to 252 electrode channels four additional analog channels and 16 digital IN OUT channels The digitized electrode data is transmitt
10. About Cardio 2D About Cardio2D L ardia2D Version 0 9 4 0 Multi Channel Systems GmbH MES Copyright MES 2008 2010 Close The dialog shows basic information about Cardio 2D Note Please keep in mind that the information of this dialog is necessary in case of support 58 Cardio 2D Analyzer Software 6 2 Control Window Cardio 2D The control window of Cardio 2D is divided in three separate windows The File information on top the Heartbeat view and the Settings file below File information E AstemizoleMEA_1 02d l2 H Heartbeat 51 41 51 32 42 52 33 43 53 34 44 54 35 45 55 36 46 56 37 057 ao 40 50 Control Window E MEAS ettings Analyze i HBDietectMade i HBTimeStampT ype H HBMinMaxMade H DetectMinRiseTime Settings file DetectDeadTime DetectSearchlnterval PropagationPath PropagationL nit Propagation T met ax MinimumClusterChannels PropagationPathElectroc H MapFieldPotential EH MapLatency 59 Cardio 2D Manual File Information C CardioZ0 D ata AstemizoleMEa_1 ced In the file information window the name of the currently analyzed Cardio 2D data or result file is displayed Additional the number of the MEA used in this experiment is represented in the file name To select the number of the recording phase you want to analyze please open the drop down menu and select the designated recording phase of that file Fo
11. MEAs are implemented The standard MEA with an 8 x 8 electrode layout grid the diameter of the MEA electrodes and the spacing between them differs a MEA with a 6 x 10 electrode layout grid the 256MEA with a 16 x 16 layout grid or the EcoFlexMEA36 with 6 x 6 electrode layout grid The design of the MEA window is automatically adjusted to the type of the MEA please see chapter Data Acquisition Settings Settings ES Cardio2D File Experiment MEAs Settings Data Acquisition File Locations Menu to define the data acquisition device and to determine the path for the data and log files 17 Cardio 2D Manual Click Data Acquisition in the Settings menu The Data Acquisition Settings dialog opens This dialog is the same which show up before starting Cardio 2D for the first time Please see above ER Data Acquisition Settings X Device LIS B MEZ5E MC Card 749 0000 OOOO Sample Rate DIA li Range 4096 mv hern Driver Version MEA 1 MEA 8 amp 8 100 10 MEA 2 MEA 8x8 100 10 MEA 3 MEA 8x8 100 10 MEA 4 MEA 8x8 100 10 Close Please enter the drop down menus to select the current data acquisition In dependency of the data acquisition device you can modify the Sample Rate and the Range The Resolution the Number of Channel and the Driver Version will be displayed Define the type of MEAs used in the experiment Click File Location in the Settings menu The Location Settin
12. W x H You can modify the size of the color map Interpolation Interpolation is a mathematic method to calculate missing information between two points of a line The bilinear interpolation uses four points of the nearest vicinity to calculate the interpolated point The bicubic method uses eight points to calculate the interpolated point The interpolation method of the nearest neighbor uses the mean value of four or five points to calculate the interpolated point 41 Cardio 2D Manual Select Data tab in Map Settings dialog MEA 1 Map Settings l Data Map Interpalatian Missing Data Nearest eighbor Missing Channel NearestMeighbor Show Time Stamp hhmmss ms hhmmss Interpolation In the Map tab of the Map Settings dialog the interpolation parameter refers to points inbetween measured data In the Data tab the interpolation parameter refers to the data generating the map Default is the interpolation method Nearest Neighbor Please see above It is possible to interpolate Missing Data or Missing Channel Show Time Stamp You can add the heartbeat time stamp to the color map using the option Show Time Stamp Different settings are offered for the actual time stamp which is displayed below the color map For displaying heartbeats for example a time stamp in milliseconds ms will be enough for potential spreading you can use the time stamp option hours minutes seconds and microseconds hhmmss ps
13. amplifiers in Port drop down menu for example COM1 defines MEA amplifier 1 COM 2 belongs to amplifier 2 and so on Press Test Connection to be sure that amplifier and the computer are correct connected Select Stim A or Stim B in menu Stimulation Enable the checkbox Blanking Select the desired stimulation electrode on the electrode field The button will appear in blue when using Stim A or in red when using Stim B Click Download for downloading the information to the MEA amplifier 32 Cardio 2D Software 4 3 3 Analyzer Settings Click Analyze icon HEI The Analyzer Settings dialog will open Click HB detection tab MEA 1 Analyzer Settings HB detection 20 Minimum Rise Time 100 ps Maximum Rise Time 1 me M Fixed Amplitude Signal Dependent 50 StdDev Detection D eadtime 100 me b Minas Type Global gt Min Max Search 10 ms 7 HB Timestamp b inimum In this dialog you define the parameter for detecting the waveform of a field potential Please see also Automated Waveform Detection and Detection Parameter The first parameter for detection is the Rise Time Select the minimum and the maximum of the rise time from the drop down menus The range involves 100 us to 2 ms for the minimum rise time and 200 ys to 5 ms for the maximum rise time Next parameter is the Amplitude You can decide to have a fixed amplitude for detection or a signal dependent amplitude The fixed amplitude
14. can click anywhere on one of the red bars and the mouse pointer will turn into a cross with four arrowheads Keep the mouse button pressed Now you can move the ROI with constant breadth to any region you want to analyze closer Again the displays in the MEA electrodes window and in the overlay plot window are adapted immediately Click with the right mouse button on one of the red bars The Edit menu appears Data iew Editor x General Fil Inside Fil Qutside Dock Name Data View 1 4 Visible v Enabled Context Menu Enabled Title Text Dataview I DI Iv UserCanedt CanFocus Color Laver 100 Gnd Lines Mirror Dataview H ame Vertical Horizontal Axes Control W Enabled Mouse Style Mone Wheel Style Anes Keyboard Style Both 63 Cardio 2D Manual MEA Electrode Field Recorded data traces are displayed in the MEA electrode field To adapt the y axis please use the drop down menu The MEA electrode field corresponds with the overlay plot window The data traces of the MEA electrodes selected in the MEA electrode field and highlighted in white are displayed in the overlay plot window in different colors ll LI A A A A a A 200 uv Time s n 500 uy E TT 1000 ps The MEA electrode field refers also to the single channel view In single channel view you define the region of interest Data of this ROI frame are shown in e
15. channels The MEA Window changes depending on the connected hardware If you connect for example an USB ME256 System for data acquisition it is possible to connect up to four MEAs You can switch between the different MEA tabs to set different setting parameters in the five views of the MEA Window independent from each other Via Control Window you start and stop the experimental protocol of the connected MEAs simultaneously MEA 1 MEA 2 MEA 3 MEA 4 U H The settings made in MEA Window influence all five smaller windows 27 Cardio 2D Manual 4 3 1 Labbook Click Labbook icon UL The dialog Labbook Settings will open Click Study tab MEA 1 Labbook Settings Study General StudylD AEI Age E13 Comment Rerecording for Cardio 20 version 1 0 0 In this dialog you fill in data referring to your experiment The Labbook Settings of MEA_1 will be stored in the settings file c2s You can save it as a template The inscriptions appear as soon as you open the template and there is no need to fill in again Click General tab in Labbook Settings MEA 1 Labbook Settings Study General Director Director Scientist Scientist Operator Operator Department Department Company Company In this dialog you fill in general data referring to your experiment You can save them as a template The inscriptions appear as soon as you open the template and there is no need to fill in again 28 Car
16. for the Cardio 2D software Please ask license multichannelsystems com You do not need a license for the Cardio 2D software if you like to test the software for getting an idea of how it works and what is possible You can test all functions run the viability test to see your data but you are not able to start an experiment Please contact license multichannelsystems com to receive a valid license key To enter the following dialog select License Key in menu Help Please type the key into the window Please enter license key and confirm the key with Set a License Please enter license key mmTETET Set Close 1 Double click the Cardio 2D exe on the installation volume The installation assistant will show up and guide you through the installation procedure 2 Follow the instructions of the installation assistant The Cardio 2D Driver is installed or updated automatically Installing the Cardio 2D Analyzer Software 1 Double click the Cardio 2D exe on the installation volume The installation assistant will show up and guide you through the installation procedure 2 Follow the instructions of the installation assistant 3 You will again be asked for a path to put the log and data files Cardio 2D does not produce raw data files result files only Raw data files created by Cardio 2D are never changed by Cardio 2D but analyzed and saved as result files It is advisable to use the same subfolders as used
17. it with wet hands Requirements for the installation Make sure that the device is not exposed to direct sunlight Do not place anything on top of the device and do not place it on top of another heat producing device Never cover the vents not even partially so that the air can circulate freely Otherwise the device may overheat 2 2 2 3 Cardio 2D Manual 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 Imprope
18. minimum FP change or faster or slower it would not be detected In other words the signal needs to cross the bottom border of the gray shading and the green shading at the same time Any signal component that matches the criteria will be detected The FP change parameter is the main parameter that should discriminate biological signals from noise The rise time parameter ensures that biological or artefact signals that meet the FP change criterion but have a shorter or longer rise time than the sodium peak are not detected 47 5 4 Cardio 2D Manual General Software Features Tracking Axes il NN ERR N N N N RR RR RE d 56 23 B 61 ba B3 b4 65 M Mclaren RM bu a Qik o i ale a 500gv 10s if ES For tracking the axis manually click on the axis and the mouse pointer will change into a hand sel To scroll the axes please move the mouse with the hand pointer For zooming the axes press the CTRL key and keep it pressed while moving the mouse or the wheel of the mouse The mouse pointer turns into a double arrow symbol A right mouse click on the axes gives four options Edit 111 Tracking Enabled Update Resume values Zoom To Fit All i In View Edit Tracking Enabled Update Rescue Values and Zoom to Fit With the command Tracking enabled you can move the axis by moving the mouse or by moving the mouse wheel in horizontal direction on the x axis and in v
19. on another cluster icon and on the MEA electrode again Clicking on the Remove icon X first and then onto a MEA electrode the electrode will be removed That means no analyzing is possible on this channel The electrode and her nearest neighbors are not displayed in the corresponding color map Settings File G MEAS ettings E MEAID Selected Layout Regularlarid Endw idth GridH eight GridSpace anidS pace arid Calumns arid Fi owg Channels nalyze apFieldPotential apLatency H HM apParameter Datalnterpolation Channellnterpolation Min Min amp uta Max MaxsAuta Maplnterpalatian T In the Settings file all settings of the currently analyzed Cardio 2D result or raw data file are displayed in a tree view The window contains all information saved during recording the file and you can read any information you need for analyzing the file It is not useful and therefore not possible to change these information in a Cardio 2D raw data file If you re analyze a raw data file with new parameters you can save the new results in a new Cardio 2D result file 61 Cardio 2D Manual 6 3 MEA Window Cardio 2D The MEA window on the right side is divided into four separate window panes and has two tabs Field Potential and Heartbeat Switching between the two tabs influences the settings of each of the four separate windows 6 4 Field Potential Tab Cardio 2D Click Field Pote
20. one recording phase The MEA electrode window corresponds directly with the color map The field potential of that single electrode is shown in MEA electrode window which is selected via mouse in the color map Select a plot of Latency or a plot of Velocity from the drop down menu in the Latency window The color map corresponds with the single channel view and shows the excitation patterns of that heartbeat marked with the blue arrow in single channel view Field Potential Heartbeat H t ep OSEE ESO us alu E Single channel view Color map Pcr m Ic NE ae all C NC laa P16 14 718 14 72 14 722 14 724 14 725 MEA electrode Recording 0 00 00 14 0300 Latency ms NE NE CES EE CEU E o 15 20 Latency Latency 53 Cardio 2D Manual Main Menu ES CardiozD Fil Tools Help File ES Cardio2D Tools Help Qpen Raw Data Open Result File Save Result File Exit Menu to open or save result files to open raw data files and to exit the program Cardio 2D generates three types of files the Raw Data file c2d the Result file c2r and the Settings file c2s Raw data file and the associated settings file always belong together in the result file the settings file is included The log file is not automatically generated but by users command The Cardio 2D Analyzer program deals with raw data and result files It is not possible to change the data of
21. traces are displayed in the MEA electrodes pane You can zoom to any channel by clicking on a channel in the MEA electrodes plate The zoomed channel is displayed in the larger single channel view The single channel view has two tabs FP field potential and HB heartbeat The heartbeat window shows either a diagram of the RR interval or of the frequency of the heartbeats The current values of the RRI or the frequency are displayed above in red numbers The heartbeat view shows the detection of a heartbeats at each electrode You can define up to four spatial clusters in case of formation of clusters with different pace makers When a heartbeat was detected the corresponding electrode appears in a deeper color The color map displays the propagation of a heartbeat from the pace maker to the outer regions of the cardiac cell culture or tissue The isochrones are indicated by black lines The colors indicate the latency distribution over the MEA Red color represents the pace maker when using the color map jet inverse The latency window shows the latency of the heartbeat between two user defined channels Please select two channels from the drop down menus As in HB tab the current value of latency is displayed above in red numbers Main Menu FA Cardio2D File Experiment ME4s Settings Help File EI Cardio2D Experiment MESS Open Template Save Template Restore Defaults Save Defaults Exit Menu
22. 1 Cardio 2D Filed potential tab 51 62 Cardio 2D Heart beat 51 70 Cardio 2D Heart beat tab 51 70 Cardio 2D Color map 51 62 70 Cardio 2D File 51 Cardio 2D Help 51 Cardio 2D Overlay plots 51 62 Cardio 2D Re analyzing 51 Cardio 2D Single channel view51 62 70 Cardio 2D Tools 51 Re analyzing esses 51 Cardiomyocytes 2 Color map window 15 40 CONGO et ladra 40 Data iati ud HO tied cetus 40 Interpolation of data 40 ION Si 40 Map Settings iii 40 interpolation of color 40 ZO MAD uri 40 Record MOVIE 11 40 COMPIESSION ili 40 Framerate acilia lion 40 Movie format AVI eeeeeeeeeeeenel 40 Conduction velocity 2 Control window 15 21 Basic n dU deus oU datas Edu 15 21 COMME E lee Giese M TUN EUR C 21 Set MaEKGE ii a 21 A A E A 21 VIabDHIty casses ist td Bassi em 15 21 Running a viability test 21 D Delay between electrodes 2 E Ecxitation 2 Error messages 77 H Heart beat window 15 39 A laicale 39 FICOUCIICY A 39 PIO E MENU ieri 39 RRIMENG rai 39 PIB 2D ii edu 39 US E R 39 L Latency window 15 38 Piot menisan 38 Log file window 15 26 M Main menu TOO Dd gs 15 Experiment i a 15 ble 15 A ai 15 MEAS uscite n dii Dots 15 Sets etc auge t UE EE 15 MEA electrode plate 15 27 Analyzer Settings
23. 10 47 gt Recording 2 Test 09 10 47 gt Recording 3 Test 09 10 53 Recording 4 Test 09 10 58 Recording 5 Test 09 11 01 gt Experiment stop 09 11 05 gt Experiment start 09 11 05 Recording 0 Test 09 11 11 gt Recording 1 Test 09 11 16 Recording Test lt 09 11 22 gt Recording 3 Test 09 11 22 gt Experiment stop 26 Cardio 2D Software 4 3 MEA Window MEA 1 Ww El la Geib E E Get 13 Ed ES ds ES ES WY ES dal Gl nl din Ga 6 Ral ina poppe 2 32 2 2 2 ie ICE Heartbeat ee ANAR Deut ie ES EN HU EN ES DN B3 cul 30 45 56 bei 75 9150 ps DELITTI ian st na ir dun cio bir lige nd Ba 00 00 00 000000 za d Do ou Single channel view Latency 500W 10s le 9 RefCh47 chas The MEA Window is divided into five window panes all showing heartbeats in different characteristics The prominent MEA electrode view displays electrode activity of all electrodes in the layout grid The Single channel view displays an user defined single channel to have an closer view The Heartbeat view shows the detection of a heartbeats at each electrode The Color map displays the propagation of a heartbeat from the pace maker to the outer regions of the cardiac cell culture or tissue The Latency view displays the latency between two user defined
24. 2 72 82 13 23 33 43 53 63 73 83 14 24 34 44 54 64 74 84 mnt PT 15 25 35 45 55 65 75 85 16 26 36 46 58 66 76 86 TU 17 27 37 57 67 77 87 28 38 48 58 68 78 Color map Control Window 1000 MEASettings Analyze HBDetectMode FORT HBTimeStampT ype Overlay plots HBMinM axM ode 1000 uv DetectMinRiseTime 00 00 00 000000 Settings file DetectDeadT ime DetectSearchlnterval PropagationPath PropagationU nit Propagation imeM ax i MinimumClusterChannels AAA Fimea le ai j lt PropagationPathElectroc MapFieldPotential Pent QAQl RO ca ile io Do ON 500p Fo KW MN EX 1000ps 4 Ja Lo E Main Window Overview The main window of Cardio 2D is build analogous to the main window of Cardio 2D On the left side is the Control Window and on the right side the MEA Window with the Field Potential and the Heartbeat tab 51 Cardio 2D Manual Control Window In the file information window on top of the left side the name of the currently analyzed file the number of the MEA and the number of the recording phase is displayed The heartbeat window thereunder shows the MEA electrodes You can zoom to any electrode by clicking on the electrode The zoomed channel is displayed in the larger single channel view It is possible to define up to four clusters in case of formation of clusters with different pace makers
25. 5 or 2 MEA Electrodes The MEA electrodes window allows real time monitoring of field potential data The electrode channels are arranged in a grid according to the MEA layout The channel identifier refers to the columns and rows of the MEA layout grid The drop down menus at the bottom of the window are for scaling the axes MEA 1 MEA 2 uw AA AAA AO OOO AA AAN A A q A AA A A AO AAA A DE mM ttt TUN EEUU TITTI NT TIT TT TTM TI CT TUTI TT ETITTTTTT 200 uv EN The electrodes number 15 and 32 without field potentials are grounded The electrode number 56 highlighted in white is the electrode which is displayed in the Single channel view Please read the next chapter 35 Cardio 2D Manual 4 3 5 Single Channel View Click field potential FP tab in single channel view 7 Ch 76 Ch 55 CH 58 CH 54 2000 1000 ssi 0 ra 1000 2000 l 3000 73 74 75 76 77 78 79 8n 81 82 Time s 500 10s 5 ud The single channel view allows a closer view to one of the MEA electrode channels You can zoom to any channel by clicking on a channel in the MEA electrodes plate The zoomed channel is displayed in the larger single channel view The plot shows the field potential uV against the time s To scale the axes please use the drop down menus The range for the field potential is 1 uV to 5000 pV the time can vary between 1 and 60 seconds Reset the axes by clicki
26. 9550 us Recording 0 00 00 17 4 20300 Latency ms dicci a a Rana i 15 20 Latency Latency The Field Potential tab is divided in four separate windows The Single channel view the MEA electrode the Latency and the Color map 70 Cardio 2D Analyzer Software Single Channel View Field Potential Heartbeat The single channel view displays the amplitude and the beating rate of all heartbeats of one recording phase Select the number of the displayed MEA electrode in the heartbeat window Single channel view Heartbeat tab and the color map correspond with each other The excitation patterns of that heartbeat marked with a blue arrow is exactly shown in the color map Selecting a new heartbeat with the blue arrow the color map is adapted immediately The user has three possibilities to analyze the heartbeats Click near to any heartbeat and it will be marked with the blue arrow This way you can jump to any heartbeat of that recording phase Or you can analyze each single heartbeat and the related color map step by step by clicking the icons Previous K and Next A Or you can run through all heartbeats automatically with Play gt and stop the performance with Stop 4j MEA Electrode 4000 000 FP uv c 2000 0 53 0 532 0 534 0 536 0 53 Time s 10 ms d In the MEA electrode window the field potential of exactly that electrode is displayed which is selected in th
27. Conventional methods measure the membrane potential that results from the electrochemical gradient directly with an intracellular electrode When ion channels are opened due to chemical or electrical stimulation the corresponding ions are moving along their electrochemical gradient In other words the resistance of the membrane is lowered resulting in an inward or outward flow of ions measured as a transmembrane current The extracellular space is conductive as well and though the resistance is very low it is not zero According to Ohm s law U R 1 the extracellular current results in a small voltage that can be measured with extracellular electrodes Extracellular signals are smaller than transmembrane potentials depending on the distance of the signal source to the electrode Extracellular signal amplitudes decrease with increasing distance of the signal source to the electrode Therefore a close interface between electrode and cell membrane is very important for a high signal to noise ratio The transmembrane current and the extracellular field potential follow the same time course The field potential is roughly equal to the first negative derivative of the transmembrane potential thus revealing an information that is comparable to conventional methods This has been shown for different types of signals derived from neuronal preparations as well as cardiac preparations This convenient method is used by Cardio 2D Simultaneous recordings of ac
28. MCS GmbH or its suppliers be liable for any special incidental indirect or consequential damages whatsoever including without limitation injuries damages for data loss loss of business profits business interruption loss of business information or any other pecuniary loss arising out of the use of or inability to use the program or the provision of or failure to provide Support Services even if Multi Channel Systems MCS GmbH has been advised of the possibility of such damages 3 1 Software Installation Installing the Cardio 2D Software System requirements Software One of the following Windows operating systems is required XP Vista or Windows 7 English and German versions supported with the NT file system NTFS Other language versions may lead to software errors Hardware Please read chapter Hardware Acquisition Systems One of the listed data acquisition hardware system MC Card USB ME Systems USB MEA256 System or USB ME32 FAI Systems is recommended by MCS Due to the amount of recorded data a computer with low performance may lead to performance problems more often therefore MCS recommends an up to date computer Please note that there are sometimes hardware incompatibilities of the data acquisition system and computer components or that an inappropriate computer power supply may lead to artefact signals Please contact your local retailer for more information on recommended computer hardware Important You need
29. Pokaan e ae aea With the help of the plot menu it is possible to modify a lot of plot parameters For example you can adapt the figure to a special layout wanted for a publication Please see also General Software Features 38 Cardio 2D Software 4 3 7 Heartbeat The heart beat window displays heart beats close to real time The detection of heartbeats of all electrode positions is monitored simultaneously That is why you have an impression of the signal propagation across the MEA In this screenshot a MEA electrode grid 8 x 8 is displayed Different layout grids are possible please see chapter Data Acquisition Settings The numbers of the electrodes refer to their position in the grid to rows and columns Using a different type of a MEA for example the layout will be adapted When the experiment is running the currently excited MEA electrodes are displayed in a deeper green color or depending of the cluster in deeper red blue or grey color So you can observe the heart beats in your cell culture or tissue in real time 00 00000 0000000 It is possible to define up to four regional clusters in case of formation of clusters with different pace makers in the cell culture or tissue Please use the Select Cluster icons aaa uu au aaa CEE gag ito lc E The clusters are marked in green blue or red color The grey cluster is undefined Click on one of the cluster icons and select the MEA electrodes referring t
30. a raw data file which was recorded with Cardio 2D But you can re record a raw data file with Cardio 2D Analyzer and save the new information in a result file Tools FA Cardio2D File Tools Help Analvze Raw Data Menu to open the Analyze dialog Click Analyze Raw Data Cardio 2D Analyzer Software Click HB detection tab in Analyzer Raw Data HB detection 20 Minimum Rise Time 100 ps Maximum Rise Time 1 ms Fixed Amplitude Signal Dependent 50 StdDey Detection Deadtime 100 ms Minhas Type Global hi hin Max Search 10 ms HB Timestamp Minimum Recording D In this dialog you define the parameter for detecting the waveform of a field potential Please see also Automated Waveform Detection and Detection Parameter Because you have to define these settings before recording data the same dialog appears in Cardio 2D MEA electrode pane Analyzer Settings The first parameter for detection is the Rise Time Select the minimum and the maximum of the rise time from the drop down menus The range involves 100 us to 2 ms for the minimum rise time and 200 us to 5 ms for the maximum rise time Next parameter is the Amplitude You can decide to have a fixed amplitude for detection or a signal dependent amplitude The fixed amplitude involves a voltage range from 10 uV up to 1 mV Using the signal dependent amplitude the waveform detection is more flexible The detection routine
31. ach channel of the MEA electrode field and in the overlay plot window If the ROI will be modified in the single channel view data traces in MEA electrode field and in overlay plot window are adapted immediately 64 Cardio 2D Analyzer Software Overlay Plot If you select one or more than one MEA electrodes in the MEA electrode field they are displayed in a overlay plot in the overlay plots window To adjust the y axis please use the drop down menu There is a prominent red bar in the diagram The function is different compared to the two red bars in single channel view limiting the ROI This red bar here defines the time stamp belonging to the currently shown excitation patterns of the color map Please expand the x axis to have a closer view to the displayed field potentials Click the icon Snapshot 7 to move the red bar time stamp step by step please click on HH the icons Step Backward and Step Forward The color map will be immediately adapted To create a movie of the excitation patterns click on the icon Create Movie A second red bar appears Now the function of the red bars is similar to the function of the red bars in single channel view The two red bars here limit the time span for the movie a region of interest too Click on the point in the middle of the bars The mouse pointer turns in an arrow with two arrowheads and you can vary the time span between the two bars Click on any other point of the red
32. actly with the stimulus pulse triggers both the stimulation and the blanking During the blanking signal both stimulus inputs are connected to the software selected stimulation electrode s while all recording electrodes are disconnected from the amplifier The last signal value on all electrode channels before the blanking pulse will be saved and then kept until the channels are reconnected to the amplifier This avoids that the amplifier gets saturated on all channels during stimulation which is the case with amplifiers without blanking circuit MEA Select MEA_Select is a software program for controlling the blanking circuit of the MEA amplifier With MEA Select it is possible to select the MEA electrode you want to stimulate Start MEA Select parallel to Cardio 2D The MEA Select main window appears MMEA select version 1 2 0 f B x Stimulation C Not Stim Ground Blanking Configuration Load za 31 41 51 61 71 12 22 32 42 se ez 72 221 10 20 5 5 sa 63 7 si sed e 14 24 34 44 54 64 74 54 Dowgloac 15 25 85 45 55 65 75 85 Change MEA 16 26 36 46 56 65 76 sel 37 el 46 se 66 17 27 37 47 57 67 77 sel e Blanking Stim Stim B wait 100 Lis Values are rounded to multiples of 20 ps LE 63 Port COMI tl Test Cann 20 56 48 56 68 75 El Al Connection OK Apply List Mode When using more than one MEA
33. al time monitoring Note Stopping the real time monitoring saves computer performance 66 Cardio 2D Analyzer Software Click the Map Settings icon The dialog Map Settings appears Click Map tab Field Potential Map Settings Data Map Color Map det Dv Invert Contour Off 100p Arrows Off b aim 1000 p Auto bl inirum 1000 pv O Auto Size fw xH 210210 2 Interpolation Bicubic ui In this dialog you have to define the settings for the color map Color Map The usual used color map for cardiac issues is the Jet Invers map but you can also choose Hot Cool or Grey maps To remove the colors from the map click Off The isochrones will remain only i A o o o e Contour The drop down menu is for changing the color of the contour for the isochrones into Black Jet Hot Cool and Grey Or you can remove the lines with Off Adjust the number of the contour lines from the numeric box beside up to 100 contour lines are possible Arrows The feature is not jet implemented in this version of Cardio 2D As soon as it will be implemented you find a Read Me on MCS web site and you can download the new version of Cardio 2D 67 Cardio 2D Manual Minimum Maximum You have to define the minimum and the maximum of the scale of the color map That means you define the time frame in which the excitation pattern are displayed in the color map Usually you start with zero in the
34. anual Welcome to Cardio 2D and Cardio 2D About Cardio 2D Program About Cardio 2D Program Important Information and Instructions Important Safety Advice Guarantee and Liability Operator s Obligations Terms of Use for the Program Limitation of Liability Software Installation Installing the Cardio 2D Software First Use of Cardio 2D after Installation Hardware Acquisition Systems Cardio 2D Software The Cardio 2D Program Overview Control Window 4 2 1 Control View 4 2 2 Log File MEA Window 4 3 1 Labbook 4 3 2 Stimulation of MEA Electrodes 4 3 3 Analyzer Settings 4 3 4 MEA Electrodes 4 3 5 Single Channel View 4 3 6 Latency Window 4 3 7 Heartbeat 4 3 8 Color Map Experimental Proceeding General Performance Noise Level Detection Parameter Automated Waveform Detection General Software Features WN mi NN O O Ul UI 11 13 15 15 21 22 26 27 28 29 33 35 36 38 39 40 45 45 46 47 48 Cardio 2D Manual 6 1 6 2 6 3 6 4 6 5 7 1 7 2 Cardio 2D Analyzer Software The Cardio 2D Analyzer Program Overview Control Window Cardio 2D MEA Window Cardio 2D Field Potential Tab Cardio 2D Heartbeat Tab Cardio 2D Troubleshooting Troubleshooting Error Messages Theoretical Background Cardiac Action Potential Appendix Technical Support Test Model Probe Contact Information Index 51 51 59 62 62 70 77 77 71 79 79 81 81 81 82 83 1 1 1 2
35. bars The mouse pointer turns in a cross with four arrowheads and you can move both bars to the section of the field potential you want to save in a movie Click the icon Create Movie m Click on the icon Start gt dle 1000 500 El ra 0 500 24 29 2d 3 24 3 Time s 1000p a E 1300 us H H While recording the movie the blue arrow indicates the progress of recording You can disrupt the movie by clicking the icon Pause Movie a The inter picture timing 1300 ps iq us allows adjusting speed and duration of a movie generated by using the above described functions 65 Cardio 2D Manual Color Map The color map displays the spreading of the excitation of a heart beat from the pace maker or the earliest time point detected if the pace maker is not on the MEA to the outer regions of the cardiac cell culture or tissue The isochrones are indicated by black lines The colors indicates the time distance between the first signal and outer regions of excitation Red color represents the pace maker when using the color map jet inverse The color map corresponds with the single channel view and with the MEA electrode field 1000 pi o4b 400 ps adm e Click the Copy to Clipboard icon Sal to copy the color map to the clipboard Click the Save Picture icon xd to save the color map as a picture in png format Click the Start icon gt to start real time monitoring Click the Pause con to stop re
36. by the Cardio 2D program 10 Software Installation 3 2 First Use of Cardio 2D after Installation Double click the Cardio 2D program icon The Data Acquisition Settings dialog appears Data Acquisition Settings Device ML Card 143 Sample Hate 20 kHz x Hange 4096 mv M Resolution Channels Driver Version mil Es C MEA 1 MEA oxo 10010 You have to select the data acquisition device if more than one data acquisition is available Select the Sample Rate the Range when using the MC Card only and the type of MEA or FlexMEA Please save these entries with the command Save Defaults Otherwise the dialog will appear each time you start Cardio 2D It is possible to change the default settings or to save several default files if you work with different configuration The dialog is also available in the Settings menu in Data Acquisition You will be asked for a path to put the log and data files The program will create subfolders for each of these file types It is advisable to put all three subfolders in a common Cardio 2D directory These settings can be changed later see Setting Options File Location par Location Settings X Haw Data C Dokumente und Einstellungen sCardia2D D ata ER Results C Dokumente und Einstellungen ardio 2D D ata ER Log Files CADokumente und Einstellungen sLardiozD D ata ER Close In case you have to reinstall the software you will be asked for a path to put the l
37. ce maker to the outer regions The MEA electrodes mirror this excitation The propagation time is the longest possible period for passing of the excitation from the first excited electrode to the last excited electrode of one cluster If the propagation time is too short you miss data for the complete heartbeat If the propagation time is too long the next heartbeat starts and the results are incorrect So the appropriate time has to be tested empirical depending on the experiment The setting for the Propagation Path is from minimum to maximum by default If you like to define a propagation path manually for example when propagation is not linear but circular please select the option Manual Path This option enables the option Set Propagation Path in the color map After selecting the option in the color map you can define a propagation path manually by clicking on the electrodes of the color map For deselecting electrodes of the path please click the electrode again Re analyzing If you change the Propagation Time in the analyzed file it is possible to re analyze the data with new parameters Click the Start icon gt The progress of recording in dependency of the different recording phases is displayed Recording 114 MANOR 57 Cardio 2D Manual Help Fie Tools Cardio2D Help About CardiazD4 Astemizole Menu for activating the online help for Cardio 2D and for displaying the dialog
38. detects the amplitude in relationship to the background noise The amplitude is calculated with its standard deviation in a range from 1 to 50 It is recommended to use a signal dependent amplitude The following parameters characterize the detected field potential Detection Deadtime defines the time span in between no further field potential should be detected The detection dead time has to be longer than the time used for searching the minimum or maximum otherwise the following error message appears AAA y The Min Max Search Interval has to shorter than the Detection Dead Time 55 Cardio 2D Manual The drop down menu Min Max Type describes the type of the minimum or maximum Local or Global These options refer to one single field potential signal Local means the first detected minimum or maximum global means the deepest minimum or highest maximum in the field potential Please see the picture 000 1000 Lm 0 ZT_ Li 1000 Local Global 2000 minimum minimum DE i LEM HS IM IESU MEC QUI GEN LUN 1 88 1 83 Time s Whether you select the local or the global minimum or maximum depends on the aim of the user If you are interested in the maximal slope of the action potential you have to select the first local minimum or maximum If you are interested in absolute minimal or maximal values you have to select the global minimum or maximum Please select the time used for searching the minimum or maximum o
39. dio 2D Software 4 3 2 Stimulation of MEA Electrodes Click Stimulate icon 1 ESMEA 1 Stimulate E x Device M CS 5TG4002 20001 Channel 1 Stimulus Syne Out Symmetric v On v 1 Phase 1000 mv Extension 300 TE 2 Phase IPI The stimulation feature gives the possibility for voltage driven stimulation of a MEA electrode This is useful for example if a cardiomyocyte culture did not form a pacemaker This dialog allows to connect a stimulus generator STG and to design the stimulation pulse You can connect either one stimulus generator STG with more than one channels or several STGs to stimulate the different MEA amplifiers Define either the channel number from the Channel drop down menu when using one STG or the devices from the Device drop down menu when using several STGs I MEA_1 Stimulate x Device MES 5 7164002 20033 MCS STG4002 20033 Channel MCS ST 4002 20041 29 Cardio 2D Manual Stimulus Pulse In the Stimulus menu you design the stimulus pulse Stimulus Symmetric yl 1 Phase 2 Phase IPI The pulse shape of a stimulus pulse is usually biphasic symmetric rectangular The parameters are the negative or positive sign of the first pulse phase the duration of each phase and the amplitude of the pulse Click the checkbox Symmetric and the pulse will automatically be symmetric and biphasic To design the pulse shapes please use the n
40. do so to learn to know the program On the Control panel click the Viability icon Data is not recorded You see the raw data streams of all 60 channels in the typical MEA layout grid You may have to adjust the position and span of the axes until you can clearly see the noise level Click any channel to zoom into this channel Following screen shot shows the typical behaviour of a MEA amplifier with test model probe 45 Cardio 2D Manual 5 2 Detection Parameter MEA 1 Analyzer Settings i HB detection 20 Minimum Rise Time 100 ps Maximum Rise Time ms Fixed Amplitude Signal Dependent 50 StdDey Detection Deadtime 100 ms gt Min Max Type Global x Min Mas Search 10 ms z HE Timestamp Minin Under the tab HB detection in Analyzer Settings of the MEA electrodes plate um you can adjust the parameters for the field potential detection Sometimes it will be necessary to play with this parameters until you find the best configuration for your experiment HB detection 20 Minimum Rise Time 100 ps Maximum Rise Time ms T FE E Signal Dependent 50 StdDev Detection Deadtime 100 ms E Min Max Type Global x Min Max Search 10 ms HB Timestamp Minimum Recording O Click Analyze Raw Data in menu Tools in Cardio 2D In the dialog Analyze you can adjust the parameters for the field potential detection 46 5 3 Experimental Proceeding The sodium curre
41. e color map via mouse To adjust the x axis please the use the up down control Click Edit Plot icon a A plot menu appears NARRAN Se UG With the help of the plot menu it is possible to modify a lot of plot parameters For example you can adapt the figure to a special layout wanted for a special publication Please see also General Software Features 71 Cardio 2D Manual Color Map 28550 us 00 00 00 509650 a H mE e Please read the chapter Color Map in Field Potential tab In Heartbeat tab an additional feature in the color map window is available Open the dialog in menu Tools Analyze Raw Data File Tools Help Analvze Raw Data 72 Cardio 2D Analyzer Software Click 2D tab in Analyzer Settings da analyze X HB detection 2O Min Cluster Channels 10 Propagation Time 20 me Propagation Path ManualPath i Minh as MM anualP ath Recording D The setting for the Propagation Path is from minimum to maximum by default If you like to define a propagation path manually for example when propagation is not linear but circular please select the option Manual Path This option enables the option Set Propagation Path in the color map Click the Set Propagation Path icon Lin the color map window After selecting the option in the color map you can define a propagation path manually by clicking on the electrodes of the color map For deselecti
42. e interpolation parameter refers to the data generating the map Default is the interpolation method Nearest Neighbor Please see above With the method of interpolation you can substitute missing channels also The program uses the same Nearest Neighbor method for interpolation If you do not want to substitute missing channels click Off In the drop down menu Show Time Stamp you can choose the type of the time stamp you want to be displayed below the color map 68 Cardio 2D Analyzer Software Click the Record Movie icon The dialog Movie Recorder appears MEA 1 Movie Recorder a Movie Type MN Frame Rate 30 Frames s Compression On E File Mame Start Cancel The movie of the color map will be recorded in AVI format The AVI format is a common format You just need a Media Player to replay the video You can choose the Frame Rate between one and hundred frames per second Select Compression of the movie with On or deselect the compression with Off Save the movie by clicking the Save icon al and select a file name and the path to the folder you want to store it Recording the color map movie starts as soon as you click Start 69 Cardio 2D Manual 6 5 Heartbeat Tab Cardio 2D Click Heartbeat tab Field Potential Heartbeat H A aM E LE EIE us D ee Single channel view Color map ia EEE pass es aa 716 14 718 14 72 14 722 14 724 14 725 MEA electrode
43. e rise time is in the defined range as well Limit the rise time range to exclude artifact signals that are in the range of the defined amplitude As the sodium peak is generally very fast small rise times such as 100 to 400 us can usually be used The higher you define the FP change parameter the higher you need to set the rise time as well as a higher FP change needs longer than a smaller one The rise time and the amplitude of the Na Peak are used to detect only signals that show the expected kinetics of the waveform Slow potential shifts are excluded Automated Waveform Detection HH 107 Min Amplitude Fo 1 Min Rise Time WA i Max Rise Time II HI 2 067 2 028 p nl wd LB ne fs This illustration shows how the Cardio 2D routine detects the fast sodium peak by the three parameters field potential change minimum and maximum rise time The software scans the recorded data points and it checks whether following data points meet the required criteria For example when the waveform detection routine arrives at the data point marked with the red dot in the figure the following data points meet the amplitude criterion shaded in gray The signal reaches this field potential change in the preset range of the rise time shaded in green The data point at which both the FP change and the rise time criteria are met is marked with a second red dot in this figure Would the signal be smaller than the
44. ed to the connected computer via universal serial bus High Speed USB 2 0 Thus it is possible to use any computer as a data acquisition computer also a laptop Recorded data is graphed analyzed and reviewed with Cardio 2D and Cardio 2D program The integrated filter amplifier supports 252 electrode channels The gain of 1100 is fixed The bandwidth of 1 Hz to 3 kHz is suitable for a broad range of applications Data Acquisition with the external USB ME32 FAI Systems The USB ME32 FAI System is a very compact and portable stand alone solution It features an integrated 32 channel filter amplifier and data acquisition The digitally converted electrode signals are transmitted to the connected computer via universal serial bus High Speed USB 2 0 As in other USB ME Systems it is possible to use any computer as a data acquisition computer also a laptop The size of the complete system is small enough to fit into a laptop bag together with the computer The system is for use with a 32 channel miniature amplifier MPA32I You can directly connect the EcoFlexMEA36 to the 32 channel miniature preamplifier for recording with Cardio 2D 13 4 Cardio 2D Software 4 1 The Cardio 2D Program Overview E Cardio2D Ol x File Experiment MEAs Settings Help ED EU SD ED Ul ii i i 42 22 32 42 52 62 72 82 adde ret a i a 43 93 33 da Ga AR 73 83 32 2 2 AAN ata 4 Heartbeat o ED OD O Gp Ge UP GE 28 38 48 58 58 78 8150 ps
45. ertical direction on the y axis Edit Click Edit and the Plot Axis Editor appears You can adjust general and specific parameters for your plot layout Plot Axis Editor x General Title Scale Display Scale Range Grid Lines Tracking Dock Min 46 4301 464997923 ax fo 261 45550424 76 Da Name Xabi 1 v Visible Title Test fTime s v Enabled Color ME Layer 100 Iw Mind E v User Can Edit v Can Focus v Controk K ey Toggle Enabled Span 20923507 0084953 Master LII Scale Type Linear Reverse Clip To Min Maz Cursor Scaler 1 48 Click the Plot icon EJ The following toolbar appears aaa ica ala Copy Picture Copy Data ni 21 1 4 2 Experimental Proceeding Tracking Scroll axes Zoom axes Zoom in and zoom out Select Zoom box Data and Cursor Edit Copy to clipboard Save Print Preview 49 Cardio 2D Manual Click Edit icon ud The following dialog appears Picture 00 7 Conto write d E Conto xm sw NENNEN Now it is possible to format the plot There are a lot of parameters available 50 6 Cardio 2D Analyzer Software 6 1 The Cardio 2D Analyzer Program Overview Cardio2D E al x File Tools Help File information Heartbeat K AstemizoleMEA_1 c2d Single channel view bi Heartbeat 21 31 41 51 61 71 12 22 32 42 52 6
46. f the field potential Min Max Search from the drop down menu range from 1 ms up to 200 ms You can set a HB timestamp to each detected field potential either to the maximum or to the minimum or to the maximal slope of the field potential This timestamp is visualized in the plot with a black dot and stored in the raw data file Please read also chapter Detection Parameter Re analyzing If you change Analyzer Settings in the analyzed file it is possible to re analyze the data with new parameters Click the Start icon gt The progress of recording in dependency of the different recording phases is displayed Recording 114 BELCE 56 Cardio 2D Analyzer Software Click 2D tab in Analyzer Raw Data da analyze X HB detection Min Cluster Channels 10 Propagation Time 20 ms Propagation Path Mind ax Mink ax MM anualPath Recording D The first drop down menu in this tab of the dialog defines the minimal number of cluster channels The Min Cluster Channels gives the minimum number of channels at which heartbeats have to be detected to form a cluster For example for a MEA electrode layout with 60 electrodes it could be 10 but if less than 60 electrodes are involved it could be 5 or 2 Please define the Propagation Time from the drop down menu in a range of 1 to 200 ms The action potential runs over the heart cell culture or tissue or hole heart from the pa
47. gs Cardia2D Help About Cardio2D Licence Key Menu for starting the Cardio 2D online help to monitor the dialog About Cardio 2D and to enter the license key For a valid license please read chapter Installing Cardio 2D Software Select About Cardio 2D ES about Cardio2D Cardio 20 Versioni 0 9 3 0 Multi Channel Systems GmbH MES Copyright E MES 2007 2010 Close The dialog shows basic information about Cardio 2D Note Please keep in mind that the information of this dialog is necessary in case of support 20 Cardio 2D Software Control Window Control view Control Window Recording 2 Test Experiment stop Experiment start Recording 0 Apply O ph Recording 1 Test O pM Recording z Apply 100 pi Recording 3 Test 100 pM Recording 4 Apply 1 nl Recording 5 Test 1 nM Recording amp Apply 10 nl Recording Y Test 10nM Experiment stop Log file The Control Window is divided into two window panes The Control View controls the experimental protocol and the Log File documents the experimental proceedings 21 Cardio 2D Manual 4 2 1 Control View In the Control window you can enter the settings for setting up and running an experiment Viability Click Viability The Viability icon Y changes into Stop icon and the duration of the viability test is counted up in hours minutes and seconds Stop the viability test manual by clicking on Stop icon Bunnit Ja 00 03 33
48. gs dialog opens ER Location Settings X Haw Data Dakumente und Einstellungen sCardia2D D ata Result LA Dokumente und Einstellungen sLardio2D D ata ER Lag Files LA Dokumente und Einstellungen ardia2D D ata EE Close Please define the file path for the raw data and the corresponding setting files the result and the log files 18 Cardio 2D Software EN Data Acquisition Settings X Device USB ME 256 bi Sample Rate 25 kHz id Range 4096 mv ha Resolution Channels 256 MEA 1 MEA 8x8 100 10 MEA 2 MEA 8x8 200 30 MEA 3 MEA 6x10_500_10 MEA 4 MEA 8x8 100 10 Close When using an USB_ME256 device for data acquisition it is possible to connect up to four MEA amplifiers which can be operated independent from each other That means you can select the settings for each MEA separately but the starting and stopping of the experimental protocol is synchronous for all connected MEAs The design of the MEA tab is different depending on the type of the connected MEA Design of the MEA window when connecting three MEAs and using a MEA with a 6 x 10 layout grid in MEA3 tab MEA_1 MEA 2 MEA_3 UA A na AD Dn A na ca co sul au m eu n B la B n GE 19 Cardio 2D Manual Design of the MEA window when using one EcoFlexMEA36 with a 6 x 6 layout grid MEA 1 Ui Ww Help EI rardin2D File Experiment MEAs Settin
49. he connected computer This information is especially important if you have modified the computer or installed new hard or software recently Test Model Probe The provided test model probe simulates a MEA with a resistor of 220 kQ kiloohm and a 1 nF nanofarad capacitor between bath and electrode for all 60 electrodes and can be used for testing the MEA amplifier Test Model Probe GND bath Pad electrode 81 9 3 Cardio 2D Manual Contact Information Local retailer Please see the list of official MCS distributors on the MCS web site User forum The Multi Channel Systems User Forum provides the opportunity for you to exchange your experience or thoughts with other users worldwide Mailing list If you have subscribed to the Mailing List you will be automatically informed about new software releases upcoming events and other news on the product line You can subscribe to the list on the contact form of the MCS web site www multichannelsystems com 82 10 Index 83 Cardio 2D Manual 84 A Automated Waveform Detection 47 B Blanking Circuit BC 36 EXTENSION ida 36 SVC QUE iolanda 36 PNIS L O 36 C Cardiac action potential 2 Cardio 2D 1 Cardio 2D Program Overview 15 Cardio 2D Analyzer 1 Cardio 2D Analyzer Program OVErVIEW ii 51 Cardio 2D Control window 51 59 Cardio 2D File information 5
50. involves a voltage range from 10 uV up to 1 mV Using the signal dependent amplitude the waveform detection is more flexible The detection routine detects the amplitude in relationship to the background noise The amplitude is calculated by the standard deviation of the signal times the factor you can choose It is recommended to use a signal dependent amplitude The following parameters characterize the detected field potential Detection Deadtime defines the time in which no detection is performed 33 Cardio 2D Manual The drop down menu Min Max Type describes the type of the minimum or maximum Local or Global These options refer to one single field potential signal Local means the first detected minimum or maximum global means the deepest minimum or highest maximum in the field potential within the Min Max search interval Please see the picture 2U00 1000 Em oO 00 Li 1000 Local Global 2000 minimum minimum DE IL A IM LES MEI a 1 06 1 83 Time sl Whether you select the local or the global minimum or maximum depends on the aim of the user If you are interested in the maximal slope of the action potential you have to select the first local minimum or maximum If you are interested in absolute minimal or maximal values you have to select the global minimum or maximum Please select the time used for searching the minimum or maximum of the field potential Min Max Search from the drop down menu ra
51. ion of Action Potential Changes from Field Potential Recordings in Multicellular Mouse Cardiac Myocyte Cultures Marcel D Halbach et al Cell Physiol Biochem 2003 13 271284 80 9 1 9 2 Appendix Technical Support Please read the Troubleshooting part of the user manual or Help first Most problems are caused by minor handling errors Contact your local retailer immediately if the cause of the trouble remains unclear Please understand that information on your hardware and software configuration is necessary to analyze and finally solve the problem you encounter If you have any question or if any problem occurs that is not mentioned in this documentation please contact your local retailer The highly qualified stuff will be glad to help you Please keep information on the following at hand Description of the error the error message text or any other useful information and of the context in which the error occurred The more information on the actual situation you can provide the easier it is to track the problem The serial number of the device You will find it on the bottom side of the main unit and on the amplifiers The software and hardware version you are currently using On the Help menu click About The displayed dialog box shows the version numbers The operating system and service pack number on the connected computer The hardware configuration microprocessor frequency main memory hard disk of t
52. lel to the Basic Settings dialog the options referring to the recording phases are available Please see above There are two series of compounds provided The dilution factor in Series 1 10 is ten and in Series 1 3 the factor three But you can create a user defined compound series by overwriting the given example in the window Please consider the syntax otherwise you get a syntax error message A A Error in Series string Please check the syntax Message Convert Dose incorrect parameter 25 4 2 2 Cardio 2D Manual In the control window beneath the Compound icon the currently applied dose of compound is displayed LS A Dose 300 pM Log File Below the control window is the log file window Each step of your experiment is documented together with its time stamp You can save the log file by clicking on the Save icon Sal The log file has the extension xml Under Settings in File Location you define the path for the log file To clear the log file display please click the Clear icon X The message Cleared Log lt Toz2 46 gt Cleared Log will appear 08 44 20 Cardio D start lt 08 44 21 gt Add MEA 1 lt 09 07 17 gt Viability start 09 09 24 gt Viability stop 09 09 28 gt Experiment start 09 09 28 gt Recording 0 Test lt 09 10 11 gt Experiment stop 09 10 30 gt Experiment start lt 09 10 30 gt Recording 0 Test 09 10 36 Recording 1 Test lt 09
53. lems that are not mentioned in this documentation occur even if you have solved the problem on your own This helps other users and it helps MCS to optimize the instrument and the documentation Please pay attention to the safety and service information chapter Important Safety Advice Multi Channel Systems has put all effort into making the product fully stable and reliable but like all high performance products it has to be handled with care Error Messages eee a x CardiazD is recording data Stop measurement before exit programm Please stop recording before exit the Cardio 2D program ie lt C Cardio D DataldefaultmMEA_1 c2d already exists Override on Please enter a new file with a new file name Be aware that Ja will overwrite present data Choose Nein to enter a dialog and save data in a new file Wie x Error in Series string Please check the syntax Message Convert Dose incorrect parameter Please check the syntax eee i 54 The Min Max Search Interval has to shorter than the Detection Dead Time Please adjust the time 77 Cardio 2D Manual 78 8 1 Theoretical Background Cardiac Action Potential Over the last 30 years non invasive extracellular recording has developed into a widely used standard method The semi permeable lipid bilayer cell membrane separates different ion concentrations charges on the inner and outer side of the membrane
54. minimum that is the pace maker The excitation pattern will run from the pace maker over the map to the maximum The widest selectable time frame for the color map spans from 0 us in the minimum to 10000 ms in the maximum To change the colors of the map please use the first drop down menu Color Map Auto Click the checkbox Auto in Maximum option The Minimum is zero you are not allowed to modify it in Cardio 2D The Cardio 2D program automatically finds for the maximum of each heart beat and adapts the scaling of the color map It is recommended to use the auto scaling function Size W x H You can modify the size of the color map Interpolation Interpolation is a mathematic method to calculate missing information between two points of a line The bilinear interpolation uses four points of the nearest vicinity to calculate the interpolated point The bicubic method uses eight points to calculate the interpolated point The interpolation method of the nearest neighbor uses the mean value of four or five points to calculate the interpolated point Click the Map Settings icon Zl The dialog Map Settings appears Click Data tab Field Potential Map Settings Data Map Interpalatian Missing Data NearestNerghbor Missing Channel Crt Show Time Stamp ps hhrmms hhmmss ms hhmmss In the Map tab of the Map Settings dialog the interpolation parameter refers to the color of the map In the Data tab th
55. ng on the Reset icon The detected heartbeats are marked with black dots The detection time stamp can be set to minimum or maximum or maximum slope in the HB Timestamp drop down menu in the Analyzer dialog Please click on the Analyzer icon in the MEA electrodes plate and enter the HB detection tab in Analyzer Settings The red line labelled with the identifier of the selected channel indicates that the zoomed MEA channel was switched Click Edit Plot icon a A plot menu appears gt nt Qik o m als With the help of the plot menu it is possible to modify a lot of plot parameters For example you can adapt the figure to a special layout wanted for a publication Please see also General Software Features Click heartbeat HB tab in single channel view The HB tab shows a diagram of the duration of the RR interval in ms if you select RRI ms from the drop down menu In hole heart experiments the RR interval is defining one complete heartbeat from one ventricular depolarization to the next ventricular depolarization 1 RR is the beating frequency of the heart So you can see ventricular tachycardia if occur In cell cultures you do not have ventricular depolarization 36 Cardio 2D Software If you select Heart Rate Hz the rate of the heartbeats is calculated from the reciprocal value of the RR interval The last actual value of the RRI ms or of the Heart Rate Hz is displayed in red ab
56. ng the experiments The offline analyzer part is called Cardio 2D The Cardio 2D Analyzer program is intended for reviewing and graphical representation of recorded experiments and for preparing the graphical results as far as possible for a publication Cardio 2D Manual About Cardio 2D Program The electrical activity that is the field potentials is recorded by the data acquisition system The software Cardio 2D maps the spreading of cardiac action potentials in cardiomyocyte cultures or tissues or via flexible MEAs on whole heart surfaces Cardio 2D pictures excitation patterns and conduction velocity of cardiac action potentials The software helps to detect re entry cycles and measures the delay between electrodes The results are represented in meaningful and impressive color maps or movies Cardio 2D Main Window Graphical User Interface GUI nx File Experiment MEAs Settings Help MEA 1 MEA 2 21 31 41 51 61 71 12 22 32 42 52 62 72 82 13 23 33 43 53 63 73 83 E OOO LI POE 14 28 30 4 S 06 78 amp 4 a ann 16 26 36 46 56 66 76 86 17 27 37 47 57 67 77 87 28 38 48 58 68 78 00 00 00 000000 Ga id P oo uw CTESTUS gt Add MEA EE ERR les LE i ER B en e VERI SY I RI fj 6450 us 14 31 17 gt Viability start lt 14 32 46 gt Viability stop 143327 Viabiiy stop MAREA RTT 14 33 33 gt Viability start Ee Ml SS AR Blam tt PITTI Ops Latency 950 us Latency ms Time s 500
57. ng electrodes of the path please click the electrode again 00 00 00 569650 73 Cardio 2D Manual Latency Instead of the overlay plot window in the Field Potential tab there is in heartbeat tab the latency window with two different diagrams Select Latency from the drop down menu Recording Ken e EE m JJ A A A da 46 Time m Latency e The diagram shows the latency ms of the excitation in dependency of the time ms between the minimum and the maximum Select Velocity from the drop down menu Recording V elacitu mm ms i NEC S MEZZE E S 44 46 Time m The diagram shows the velocity mm ms of the excitation in dependency of the time ms between the minimum and the maximum Click Edit Plot icon 3k A plot menu appears ont 48 SG UGG With the help of the plot menu it is possible to modify a lot of plot parameters For example you can adapt the figure to a special layout wanted for a special publication Please see also General Software Features 74 Cardio 2D Analyzer Software 75 7 1 7 2 Troubleshooting Troubleshooting Most problems occur seldom and only under specific circumstances In most cases it is only a minor problem that can be easily avoided or solved If the problem persists please contact your local retailer The highly qualified staff will be glad to help you Please inform your local retailer as well if other prob
58. nge from 1 ms up to 200 ms You can set a HB timestamp to each detected field potential either to the maximum or to the minimum or to the maximal slope of the field potential This timestamp is visualized in the plot with a black dot and stored in the raw data file Please read also chapter Detection Parameter Click 2D tab in Analyzer Settings 1 MEA 1 Analyzer Settings X HB detection 2O Propagation Time 10 ms Min Cluster Channels 34 4 3 4 Cardio 2D Software In this dialog you can define the Propagation Time from the drop down menu in a range of 1 to 200 ms The action potential runs over the heart cell culture or tissue or hole heart from the pace maker to the outer regions The MEA electrodes mirror this excitation The propagation time is the longest possible period for passing of the excitation from the first detected excited electrode to the last detected excited electrode of one cluster If the propagation time is too short you miss data for the complete heartbeat If the propagation time is too long the next heartbeat starts and the results are incorrect So the appropriate time has to be tested empirical depending on the experiment The Min Cluster Channels gives the minimum number of channels at which heartbeats have to be detected to form a cluster For example for a MEA electrode layout with 60 electrodes it could be 10 but if less than 60 electrodes are involved it could be
59. nt is detected by the rapid change in the field potential indicated by the rise time The rise time refers to the time required for a signal to change from a specified low value to a specified high value Typically these values are 10 and 90 of the step height The output signal of a system is characterized also by fall time If the rise time is chosen very short the slope of the peak is very steep if the rise time is longer the slope will be smooth You have to try out which time is the best to characterize the Na Peak you want to detect You can either define a fixed value for the amplitude of the field potential for example 100 uV Using the signal dependent amplitude the waveform detection is more flexible The detection routine detects amplitudes in relationship to the noise level The amplitude is calculated with its standard deviation in a range from 1 to 50 It is recommended to use a signal dependent amplitude The standard deviation is used to estimate the individual amplitude change for each channel separately A time interval of 1 s is used to calculate the standard deviation You set the factor by which the standard deviation is multiplied 1 to 50 To compensate individual differences in the noise level of channels it is recommended to use the signal dependent amplitude as detection parameter The data is scanned for a situation where the minimum amplitude either fixed or relative to the standard deviation is met and th
60. ntial tab Field Potential Heartbeat m i m Single channel view y Color map 1000 us 200 py 14079 ms 1000 yv Overlay plot T 14 08 Time s E KH H gE 1000ps The Field Potential tab is divided in four separate windows The Single channel view the MEA electrode field the Overlay plot and the Color map 62 Cardio 2D Analyzer Software Single Channel View Field Potential Heartbeat The single channel view displays the field potentials of one recording phase of the in the heart beat window selected MEA electrode With the Zoom Box icon you can zoom to any region of the data trace Click the Reset Axes icon to reset the default settings of the axes Click Edit Plot icon 3k A plot menu appears gt nt QAQlko Mla alS With the help of the plot menu it is possible to modify a lot of plot parameters For example you can adapt the figure to a special layout wanted for a special publication Please see also General Software Features The two red bars in the heart beat plot of the single channel view represent the region of interest ROI Click on the middle of one red bar near to the zero line The mouse pointer will turn into an arrow with two arrowheads Keep the mouse button pressed Now you are able to vary the breadth of the ROI The displays in the MEA electrodes window and in the overlay plot window are adapted immediately Additional you
61. o this cluster by clicking on the electrode You can deselect or overwrite the MEA electrodes by clicking on another cluster icon and on the MEA electrode again Clicking on the Remove icon x first and then onto a MEA electrode the electrode will be removed That means this channel is excluded from recording The selected clusters are mirrored in the color map window Each cluster is displayed separately one after the other Click the Start icon b to start real time monitoring Click the Pause icon Au to stop real time monitoring manually Note Stopping the real time monitoring saves computer load 39 Cardio 2D Manual 4 3 8 Color Map The color map displays the propagation of each heart beat across the MEA The isochrones are indicated by black lines in this screenshot but the user can change their color in Contour The colors map is represented as false color Below the map the actual time stamp is displayed Click the Copy to Clipboard icon Hal to copy the color map to the clipboard Click the Save Picture icon zd to save the color map as a picture in png format Click the Start icon gt to start real time monitoring Click the Pause con A to stop real time monitoring Note Stopping the real time monitoring saves computer load Click the Map Settings icon M The dialog Map Settings appears Select Map tab MEA 1 Map Settings Data Map Color Map Jet el Invert Contour Black
62. og and data files again Note It is recommended to save the settings in the Data Acquisition Settings dialog as default This way you do not have to type them in again when restarting Cardio 2D 11 Cardio 2D Manual For saving the settings as default please enter File menu Save defaults ES Cardio2D MESSO Experiment Open Template Save Template Restore Defaults Save Defaults Exit 12 3 3 Software Installation Hardware Acquisition Systems Data Acquisition with the internal MC_Card The data acquisition computer with the MC_Card comes preinstalled and preconfigured by MCS for a flawless operation You should contact your local retailer for assistance if you want to install additional hard or software or if you want to replace the computer as incompatibilities of hardware components or software settings with the MC_Card or Cardio 2D may occur Caution You have acquired a high performance data acquisition and analysis computer Do not modify the system do not install new hard or software or another operating system without asking MCS or your local retailer for advice Especially do not install virus scanners or firewalls because these programs are known to interfere with the data transfer to the hard disk MCS cannot guarantee that a modified system is fully operational Even data loss may occur preconfigured by MCS and should not be changed by the user Changing these
63. oratory 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 High Voltage Electrical cords must be properly laid and installed The length and quality of the cords must be in accordance with local provisions Only qualified technicians may work on the electrical system It is essential that the accident prevention regulations and those of the employers liability associations are observed Each time before starting up make sure that the mains supply agrees with the specifications of the product Check the power cord for damage each time the site is changed Damaged power cords should be replaced immediately and may never be reused Check the leads for damage Damaged leads should be replaced immediately and may never be reused Do not try to insert anything sharp or metallic into the vents or the case Liquids may cause short circuits or other damage Keep the device and the power cords always dry Do not handle
64. ording 0 Apply D mit Data 11 19 25 gt Recording 0 Test 11 13 31 gt Recording 1 Apply Orit Data lt 11 20 56 gt Experiment start lt 11 13 34 gt Recording Tes 11 20 56 gt Recording 0 Apply z 11 19 44 gt Recording 2 Apply Orit Data 11 21 01 gt Recording 1 Test lt 11 19 46 gt Recording 2 Test 11 21 02 gt Recording 2 Apply lt 11 19 55 Recording 3 Apply Omit Data 11 21 09 Recording 3 Test lt 11 19 59 Recording 3 Test z 11 21 11 gt Recording 4 Apply lt 11 20 04 gt Recording 4 Apply Omit Data lt 11 21 14 Recording 5 Test lt 11 20 14 gt Recording 4 Test 11 21 17 gt Experiment stop z 11 20 17 gt Experiment stop Click Complete and all data of this recording phase will be saved in the raw data file Click Omit Data and the raw data recorded in the time frame you need to type a comment and to start with Set Marker again is omitted from raw data file A new entry in the log file of the Log file window appears 24 Cardio 2D Software Click Compound in Experiment The Basic icon is replaced by the Compound icon The Compound Settings dialog appears Compound Settings f Oph 100 phi 1 nM 10 nbi 100 nM 1 phi 10 p Click the Compound icon Switch Recording Clicklcon Recording Complete Close In this dialog you can choose series of compounds which are applied to the cardiac cells or tissue on the MEA In paral
65. ot have to type in them again Stimulus Generator STG It is recommended to use a stimulus generator STG of the 2000 or 4000 series from Multi Channel Systems MCS GmbH Important When working for example with four amplifiers with blanking circuit BC you have to connect a STG4004 or a STG2004 The TTL blanking signal and the stimulation pulse must be synchronized that is you have to connect the stimulus pulse of channel 1 on MEA amplifier 1 to the TTL blanking pulse of Sync Out 1 of the STG channel 2 to Sync Out 2 channel 3 to Sync Out 3 and channel 4 to Sync Out 4 respectively When using an USB_ME256 device for data acquisition you can connect up to four MEA amplifiers You define the stimulation settings for each of the connected MEA amplifier independent from each other and you can start and stop the stimulation independent from each other Amplifier without Blanking Circuit BC The stimulation electrode is manual selected directly on the amplifier via connector pins around the MEA You will see the answer of the stimulation during data recording because of the stimulus artefacts and the spreading of the field potential It does not matter whether the Sync Out checkbox in the stimulation dialog is selected or not it will have no effect 31 Cardio 2D Manual Amplifier with Blanking Circuit BC The blanking feature is used to reduce stimulus artefacts in MEA recordings A TTL pulse the blanking signal that has to be timed ex
66. ove the plot FP HB RRI ms 626 FOD E BS DE E rege cocto tt e t tetti CARA SERA A SA 10 15 2n a Time s RRI ms i FP HB HeartRate 1 59 Td s 1 5 ecesetsetest eet t Tho si La 1 4 PPT E a gp EE CIE 2 10 15 Time s HeartRate op Click the Edit Plot icon 3k A plot menu will open gt nta age B s geo With the help of the plot menu it is possible to modify a lot of plot parameters For example you can adapt the figure to a special layout wanted for a special publication Please see also General Software Features 37 Cardio 2D Manual 4 3 6 Latency Window Ref Ch 52 Lh 48 o aoo o o o d n o B b He Oe d 4444 4t B 5 B B 4 i Den T a E MO Time s The diagram of Latency window shows the latency ms of the excitation between two user defined electrodes as a function of the time s Select two electrodes from the drop down menus on the right The first selected electrode is the reference electrode the latency to the second selected electrode will be calculated The red line indicates that the selected MEA electrodes have been changed The identifiers of the channels are displayed in red The left drop down menus are for scaling the axes The range for the latency axis involves 1 ms to 500 ms the time axis can vary between 1 and 60 seconds Reset the axes by clicking on the Reset icon Click Edit Plot icon 3k A plot menu appears
67. r analyzing one recording wl phase after the other you can use the icons to switch step by step forward or backward Heartbeat Cardio 2D O X TH cb HH oce ds The heartbeat window is exactly build like the heartbeat window in Cardio 2D but the function is a little bit different It also shows the MEA electrodes in the electrode grid You can zoom to any electrode by clicking on the icon Show Channel Data e and on the electrode The zoomed electrode is displayed in the larger single channel view But it is not possible to monitor heart beats in real time that is why the Start m and Pause icons are disabled In this screenshot a MEA electrode grid 8 x 8 is displayed Different layout grids are possible please see chapter Data Acquisition Settings The numbers of the electrodes refer to their position in the grid to rows and columns Using a different type of a MEA for example the layout will be adapted You can define up to four regional clusters in case of formation of clusters with different pace makers in the cell culture or tissue 60 Cardio 2D Analyzer Software aaa gaa gag aaa gag gag aaa au uuu Please use the Select Cluster icons E The clusters are marked in green blue or red color The grey cluster is undefined Click on one of the cluster icons and select the MEA electrodes referring to this cluster by clicking on the electrode You can deselect or overwrite the MEA electrodes by clicking
68. rly 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 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 4 2 5 Important Information and Instructions Terms of Use for the Program You are free to use the program for its intended purpose You agree that you will not decompile reverse engineer or otherwise attempt to discover the source code of the software Limitation of Liability Multi Channel Systems MCS GmbH makes no guaranty as to the accuracy of any and all tests and data generated by the use the software It is up to the user to use good laboratory practice to establish the validity of his findings To the maximum extent permitted by applicable law in no event shall Multi Channel Systems
69. se automatically on Timer or you can switch to a new recording phase manually Select Click Icon from the drop down menu if you want to start a new recording phase manual il by clicking on the icon m Select Timer if you like to start a new recording phase automatically on timer and please define the phase duration 1 to 30 min from the drop down menu The number of the recordings are counted up and displayed qu Reca ding a 23 Cardio 2D Manual Select the option Enter Comment if you like to write comments concerning the different phases of recording The comments will be saved in the raw data settings and result files They are available in Cardio 2D If the option Enter Comment is selected each time when a new recording phase starts the dialog Set Marker will show up and you can type in your comment With the command Set Marker the new recording phase starts A new entry in the log file of the Log file window appears Comment Hifedipin Set Marker Basic Settings Switch Recording Timer x Phase Duration 1 min Enter Comment Recording Complete E During the time you need to type in comments in the Set Marker dialog and to start a new recording phase with the command Set Marker you can deal with the recorded data in this time frame different You can save the recorded data or you can cancel them Click Complete Click Omit Data 11 13 18 Experiment start 11 13 18 Rec
70. tion potentials with intracellular electrodes and field potentials with extracellular electrodes have shown that there is a linear relationship between the rise time of the cardiac action potential AP and field potential FP as well as between AP and FP duration The contribution of different ionic transmembrane currents can be identified in the shape of the FP waveform as well as shown in the following picture The correlation between the waveform components and the ion channel activities was shown by using ion channel blockers or depleting the medium of the respective ions You can clearly see the rapid component of the depolarizing sodium current and the slow calcium current 4 2000 Na 1000 Em zL T LL 1000 2000 ii iii ee MEC MEME 1 88 1 83 Time s The delayed rectifying K current IKr is represented either by a positive or negative peak The polarity of the peak depends on several parameters for example the proximity of the cell layer to the measuring electrode and cannot be predicted which does not matter for this assay The field potential duration corresponds to the action potential duration which can be correlated to a QT interval in an electrocardiogram It is measured from minimum of the Na peak to the maximum minimum of the IKr current peak 79 Cardio 2D Manual Electrocardiogram ST Segment ORS Complex PR ORS ST Interval Interval Interval OT Interval Reference Estimat
71. to have installed the latest data acquisition driver to operate the device which is automatically installed with Cardio 2D The installation may be invalid if the data acquisition does not respond Please contact Multi Channel Systems MCS GmbH or your local retailer in this case Recommended operating system settings The following automatic services of the Windows operating system interfere with the data storage on the hard disk and can lead to severe performance limits in Cardio 2D These routines were designed for use on office computers but are not very useful for a data acquisition computer Turned off Windows System Restore Windows Indexing Service deselected for all local disks Optimize hard disk when idle automatic disk fragmentation turned off It is also not recommended to run any applications in the background when using Cardio 2D Remove all applications from the Autostart folder Important Please do not run MC Rack software program from Multi Channel Systems MCS GmbH in parallel to Cardio 2D software Please check the system requirements before you install the software MCS cannot guarantee that the software works properly if these requirements are not fulfilled Important Please make sure that you have full control over your computer as an administrator Otherwise it is possible that the installed software does not work properly Cardio 2D Manual Installing the Cardio 2D Software Make sure to have a valid license
72. to open or save templates to restore and save defaults and to exit the program If you design experiments for a certain purpose and you want to reuse the experimental schedule please save the file as a template You can reuse the template file as often as necessary You can save as much templates as needed Factory set defaults are the setting parameters which are used when starting Cardio 2D for the first time Please enter your own defaults for example in Data Acquisition Settings dialog after installing the software Click Save Defaults to save your settings as a default This way your setting parameters will be present each time you start Cardio 2D To restore default settings please select Restore Defaults 16 Cardio 2D Software Experiment ES Cardio2D Experiment Basic Compound Menu to schedule an experiment Selecting Basic or Compound changes the icons in the Control Window Basic defines the default settings for an usual experiment Compound gives the possibility to apply an user defined series of compounds MEAs ES Cardio2D Settings Help MEA 1 MEA 8x8 200 30 MEA 2 MEA Bx8 100 10 Menu to select or deselect a MEA for recording Depending on the hardware up to four MEAs are available Enable the checkbox to select a MEA If the checkbox is disabled the MEA tab will be removed from the display and no data are recorded File Experiment Four different types of
73. umeric updown controls Phase 1 and Phase 2 The IPI the inter pulse interval is the time frame between two pulses in sequence The red line indicates the stimulus pulse the blue line indicates the TTL blanking pulse Stimulation Pulse Channel 1 Inter Pulse Interval TTL Pulse for Blanking Signal from Sync Out Sync Out Channel 1 Extension Extension 30 Cardio 2D Software Blanking Pulse The Sync Out menu is for the blanking feature if an amplifier with blanking circuit is connected Sunc Out Un m Extension 300 Us When using the blanking feature please enable the Sync Out checkbox You have to define the time of Extension The TTL pulse for the blanking signal starts synchronal to the stimulus pulse but it should last longer to avoid stimulus artefacts This time interval is called extension time The extension time varies between 0 and 1000 us the default is 300 us In some cases it may be necessary to see the stimulus artefacts Please deselect the checkbox Sync Out to switch off the Sync Out signal Additional please select the blanking feature in the Blanking checkbox of the MEA Select program see chapter MEA Select Blanking Blanking Click the Start icon gt If your experimental schedule will always need the same pulse paradigm please store the settings in menu File Save Defaults This way the settings appear in a new file and you do n
74. uv 7 10s i Y RefCh47 Ch48 1 4 Introduction About Cardio 2D Program The Cardio 2D program is an analysis software for enhanced analysis and visualization Cardio 2D Main Window Graphical User Interface GUI Cardio2D E 2 0 x File Tools Help K AstemizoleMEA_1 c2d 21 31 41 51 61 71 12 22 32 42 32 62 72 82 13 23 33 43 53 63 73 83 14 24 34 44 54 64 74 84 15 25 35 45 55 65 75 85 16 26 36 46 56 66 76 85 17 27 37 57 67 77 87 28 38 48 58 68 78 Control Experiment 1000 uv Stimulate 200 uv DataSource DeviceBus lt DeviceName i VendorlD i ProductlD BCDDevice lt SerialNumber gt lt Driverversion gt HwChannels InputR ange Gain Resolution SampleR ate MEASettings Analyze MapFieldPotential MapLatency l MovieFieldPotential Time s MovieLatency 500 pV EE 1000us 4 I8 E 8 Important Information and Instructions 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 lab

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