Multichannel-systems Roboocyte2 Manual Manuel d'utilisateur

Roboocyte2 Manual

Roboocyte2 Manual 96 Data Display Whenever you select one or more recordings - up to all recordings in the plate - in the "Data List", the

Analysis with Roboocyte2+ 97 Drift Correction The two cursors appearing in green color are used for correcting a (linear) drift of the holding curren

Roboocyte2 Manual 98 Changing Time Unit To change the time unit of the result display, please use the "Time Unit" drop down menu. Cursor T

Analysis with Roboocyte2+ 99 Result Display On the right side of the lower part you can choose in the "Result Display" whether the recording

Roboocyte2 Manual 100 When these conditions are fulfilled, data will be plotted in the Result Display. When you select one recording, one concentrati

Analysis with Roboocyte2+ 101 Capacitive Peak Currents Below the “Result Display”, there are a number of icons changing the appearance o

Roboocyte2 Manual 102 Result Display: Dose-Response Curve Click on the "Try to fit a Logistic Curve on the Data Points" icon to open the d

Analysis with Roboocyte2+ 103 Click on the button to open a window displaying the start parameters used for the actual dr-fit. When traces from mo

Roboocyte2 Manual 104 Result Display: IV-Curve Similarly, IV-recordings can be displayed individually, or in a set of IVs which can be plotted simult

Analysis with Roboocyte2+ 105 After selecting AND the IV-dependency becomes corrected for the linear leak and becomes "flat" as expecte

7 2 Important Safety Advices 2.1 Important Safety Advice Warning: Make sure to read the following advice prior to installations of the Roboocyte2.

Roboocyte2 Manual 106 Display Tools Scaling the Axes Similar as with the displays in the Roboocyte2 program, you can scale and zoom the “Data Displa

Analysis with Roboocyte2+ 107 8.4 Data Export to ASCII Format The raw data, recorded with Roboocyte2 software, is saved in the plate "*.rpf"

Roboocyte2 Manual 108 Browse through the folders to define the path for the export file. Switch from the window "Available Columns" the des

Analysis with Roboocyte2+ 109 Exporting Parameters from List View Table To export any of the parameters listed in the “List View” table, select "

Roboocyte2 Manual 110 The dose-response curve calculated from the fit-parameters above which will be saved under the file name name_CalculatedFit.da

111 9 Appendix 9.1 Contact Information Local retailer Please see the list of official Roboocyte2 distributors on the MCS web site. User forum The Mul

Roboocyte2 Manual 8 2.1.3 Compressed Air Supply Even small amounts of water in the compressed air can lead to a corrosion of the carrier. Other cont

Important Safety Advices 9 2.1.7 Regular Backups You (or the administrator) should perform backups of the Roboocyte2 data files (*.rcd files) and of

11 3 Setting Up Hardware and Software 3.1 Setting up Roboocyte2 Hardware and Software Connect all cables as described below. Warning: Carefully la

Roboocyte2 Manual 12 Connecting the Carrier 1. Plug the carrier connector into the according socket located on the Roboocyte2's rear panel. 2.

Setting Up Hardware and Software 13 3.2 Setting up the Roboflow-System The Roboflow-System has been especially designed to work with Roboocyte2. The

Roboocyte2 Manual 14 Attaching the Manifold Please control the provided manifold, specially the connections of the provided tubes to the manifold. If

Setting Up Hardware and Software 15 Attaching the Tubing to the Valve Pump Open the screw of the press bracket of the valve pump and remove the p

Information in this document is subject to change without notice. No part of this document may be reproduced or transmitted without the express w

Roboocyte2 Manual 16 Software Control for Inserting the Tube Section (PPRT1.14( in the Valve Pump Please switch to the "Manual Mode" in Rob

Setting Up Hardware and Software 17 Attaching the Tubing of the Waste Pump Connect the tubing to the provided connectors of the waste pump (PPRT2.29).

Roboocyte2 Manual 18 3.3 Setting Up the Gilson GX-271 Liquid Handler For high-throughput screens, the Roboocyte2 can be used with an industry standa

Setting Up Hardware and Software 19 General Setup of the GX-271 Liquid Handler 1. Unpack and set up the Minipuls peristaltic pump (MP3) and the Gils

Roboocyte2 Manual 20 1. Set the z-arm to the height of 175 mm by using the ruler integrated into the z-arm, marked by a blue arrow in the preceding

Setting Up Hardware and Software 21 Electrical Connections Please note that the Roboocyte2 needs to be connected to the Gilson GX-271 via the Gilson 5

Roboocyte2 Manual 22 Enabling the Gilson Liquid Handler GX-271 in the Roboocyte2 Program After installing the Roboocyte2 program, enable liquid handl

Setting Up Hardware and Software 23 3.4 Installing the Software Roboocyte2 The operating system Microsoft Windows ® 7 (32 and 64 bit) or XP (with Serv

Roboocyte2 Manual 24 3.5 Testing the Roboocyte2 Each delivered Roboocyte2-System has undergone extensive tests at the MCS site. Please follow the pr

Setting Up Hardware and Software 25 3.5.2 Testing the Roboflow Valves and Pumps The Roboflow was designed to work with the Roboocyte2 and is therefor

iii Table of Contents Introduction 1 About this Manual 1 Welcome to the Roboocyte2 2 Important Safety Advices 7 Important Safety Advice 7 High V

Roboocyte2 Manual 26 3.5.3 Testing the Amplifier with the Test Model Cell You can use the provided test model cell (TMC) to test the integrated digi

Setting Up Hardware and Software 27 Continuous Recording Switch to the "Measuring" tab page of the "Manual Mode" window. Start th

Roboocyte2 Manual 28 Now the voltage trace should exactly run at -60 mV and the holding current should be close to -600 nA (because the "oocyte

Setting Up Hardware and Software 29 Press the two autoscale arrows at the bottom of the data plot window . This will bring the current traces compl

Roboocyte2 Manual 30 Testing the Noise Level The current noise level during voltage clamp can be evaluated by recording from the TMC at different sam

Setting Up Hardware and Software 31 Decreasing the sampling rate will decrease the current noise level, increasing the proportional gain will increase

33 4 Roboocyte2 Software 4.1 Introduction Operate the Roboocyte2 robot, collect and evaluate the data by using the Roboocyte2 and Roboocyte2+ softwar

Roboocyte2 Manual 34 4.2 Main Window of Roboocyte2 Software After starting the Roboocyte2 software you will see the main window with its four differ

Roboocyte2 Software 35 The Menu and Tool Bar Use the different settings available in the menus to operate all functions of the Roboocyte2 software. S

Roboocyte2 Manual iv Recording with Roboocyte2 Control Software 69 Preparations for Recording 69 Alignment 72 Testing the Electrode Resistance 7

Roboocyte2 Manual 36 Settings Menu Change the paths for scripts, plates, or liquids and open the options window via the "Settings menu". C

Roboocyte2 Software 37 Tools Menu In "Tools" menu you can open the manual mode window, select an existing or define a new liquid configurat

Roboocyte2 Manual 38 Help Menu Open the Roboocyte2 "Help" menu to activate the online "Help" or the show the "About"

Roboocyte2 Software 39 The Tool Bar Clicking on the icons in the tools bar gives you easy access to often-used functions. In addition, you can access

Roboocyte2 Manual 40 2. Load a well plate generated by the Roboinject program Chose "Load Plate" from the file menu to load a an existing w

Roboocyte2 Software 41 Entries in red are well plates stored in the Roboinject folder (by default) or in the specified plate folder. Entries in blue a

Roboocyte2 Manual 42 Using the mouse for scaling Instead of entering numbers to scale the axis, you can use the mouse to move or scale axes. Move the

Roboocyte2 Software 43 4.2.3 The Script or Log Window Like the well plate view window, the script or log window is empty after the program start. Th

Roboocyte2 Manual 44 The script editor will open. For details please refer to the chapter "Writing a script with the built-in script editor&qu

Roboocyte2 Software 45 Click "Show Plates" button to list the plates already measured. Click "Create new empty database" to creat

1 1 Introduction 1.1 About this Manual This manual comprises all important information about the first installation of the Roboocyte2 hardware and s

47 5 Manual Mode 5.1 About the Manual Mode Although, the main purpose of the Roboocyte2 is the automated mode, almost all functions can be accessed

Roboocyte2 Manual 48 General Advice Warning: Do not use any movement command without a valid alignment. Always make sure that a proper alignment has

Manual Mode 49 Functions in the "Plate Carrier" Section All functions in the Plate Carrier section refer to functions of the carrier. Clic

Roboocyte2 Manual 50 Click the "Check" button in the "Resistance" window to display the actual electrode resistances, which will

Manual Mode 51 5.3 Manual Mode Measurement The "Measurement" tab page of the "Manual Mode" The "Measurement" tab page o

Roboocyte2 Manual 52 The upper section consists of command buttons for movements, amplifier controls before impalement and valve and pump control but

Manual Mode 53 The IV Protocol editor The lower part of the "Measurement" tab is dedicated to IV "Voltage Step Protocol" recording

Roboocyte2 Manual 54 IV Protocol Click "Create" button to create a new voltage step protocol in the "Voltage Step Protocol" dialo

Manual Mode 55 The example "TMC test" is composed of three columns starting with a step from 0 mV to -100 mV and back to 0 mV. to vary the n

Roboocyte2 Manual 2 1.2 Welcome to the Roboocyte2 The Roboocyte2 is a fully-automated all-in-one solution for high-throughput screenings of ligand-

Roboocyte2 Manual 56 5.4 Manual Mode Liquid Handling The "Liquid Handling" tab in "Manual Mode" gives you access to all function

Manual Mode 57 Roboflow-System Connect the Roboflow-System from Multi Channel Systems MCS GmbH and switch it on. Please refer to the chapter "Se

Roboocyte2 Manual 58 The Gilson Liquid Handler GX 271 If a Gilson Liquid Handler is connected and switched on, the "Liquid Handler" part o

Manual Mode 59 Selecting Racks and Tubes Select a certain rack from the "Slot" drop down menu and a "Tube" from the respec

61 6 Compound Application 6.1 Linking Compound Applications to Recorded Data For later evaluation of your recorded data, it is essential that all in

Roboocyte2 Manual 62 Roboflow-System Click the button "Valve Liquids" to open the "Composition of Compounds per Valve" editor fo

Compound Application 63 Defining a Compound Click on the compound icon to open the compound name editor "Edit Compound". Please define in

Roboocyte2 Manual 64 A right-click on the column header "Compound" opens the "Autofill Concentration" dialog for choosing the de

Compound Application 65 6.2 Using the Roboflow System The Roboflow-System is simple, easy to maintain, and ideally suited for most rapid standard and

Introduction 3 Operation Summary  The integrated TEVC amplifier operates in voltage and current clamp mode.  Instead of having a vast amount of di

Roboocyte2 Manual 66 For often needed solutions like Ringer's solution or reference compounds, it is recommended you use the transfer ports of t

Compound Application 67 GX-271 Rinse and transfer port station. Important: Please note that only the leftmost rinse, drain, and transfer ports can be

69 7 Recording with Roboocyte2 Control Software 7.1 Preparations for Recording General Preparations Either you plan to do recordings in manual mode

Roboocyte2 Manual 70 6. Click the alignment icon in the main window. The "Preparation" window of the "Alignment Wizard" dialo

Recording with Roboocyte2 Control Software 71 Installing the Well Plate Click the alignment icon in the main window. The "Preparation" wind

Roboocyte2 Manual 72 7.2 Alignment You have to align the position of the carrier relative to the measuring head i.e. the electrode tips under microsc

Recording with Roboocyte2 Control Software 73 Click next to open the "Alignment" window and the carrier will move to the x/y coordinates of

Roboocyte2 Manual 74 If you need to start the alignment again, for example if you have moved the carrier accidentally by hand, click "Cancel&quo

Recording with Roboocyte2 Control Software 75 Click on the "ResistancePos" button to move the carrier to the "Resistance Check Position

Roboocyte2 Manual 4 The integrated pinch valve Roboflow-System is an ideal perfusion system for drug receptor characterizations and quick expression

Roboocyte2 Manual 76 Gilson Liquid Handler When using the Gilson Liquid Handler, the manifold will not be used and the tubing coming from the Gilson

Recording with Roboocyte2 Control Software 77 7.5 Recording Data Manual Recording Mode What You “Can Do”:  moving the measuring head into wells  m

Roboocyte2 Manual 78 4. Start the recording in current clamp mode with Clamp Current 0 nA. This will display the voltage of Current and Voltage elec

Recording with Roboocyte2 Control Software 79 If the impalement was successful, you will notice a change in voltage on both electrodes, for example

Roboocyte2 Manual 80 6. Stop the current clamp recording and start the recording under voltage clamp by selecting "Voltage" instead of &qu

Recording with Roboocyte2 Control Software 81 In addition, you can execute IV protocols to work with voltage gated ion channels or test the voltage d

Roboocyte2 Manual 82 7.6 Writing a Script with the built-in Script Editor Please load a script via main menu "Script", "Load Script&q

Recording with Roboocyte2 Control Software 83 Although you can use every text editor for reviewing, editing or writing a script, MCS recommends to use

Roboocyte2 Manual 84 When you continue typing, for example Robo2.Sto... the command is selected in the list box and you can select it with a mouse c

85 8 Analysis with Roboocyte2+ 8.1 The Roboocyte2+ Analysis Software 8.1.1 Analysis with Roboocyte2+ The Roboocyte2+ program is the offline analyzer

Introduction 5 The Roboocyte2 Software Operate the Roboocyte2, collect and evaluate the data by using the Roboocyte2 software. The easy-to-use graphi

Roboocyte2 Manual 86 Roboocyte2+ Main Window The main window of the Roboocyte2+ software is divided in an upper section with the "Tree View&quo

Analysis with Roboocyte2+ 87 Menu to "Open Plate" to introduce a Roboocyte2 raw data file (*.rpf) into the analysis software. The last used

Roboocyte2 Manual 88 8.2 Selecting a Database and Loading a Plate File In Roboocyte2 data acquisition software a raw data file is generated "*.

Analysis with Roboocyte2+ 89 Loading a Plate File from the File System Click main menu "File", "Open Plate from the File System" t

Roboocyte2 Manual 90 8.3 Analyzer Displays In Roboocyte2+ program, you can either open a plate via "File" menu or by clicking on the "

Analysis with Roboocyte2+ 91 Data List The selection in the “Tree View” section will determine the displayed wells and recordings in the “Data List”

Roboocyte2 Manual 92 In order to have a closer look at the recordings in the “Data List”, click on the triangle icon "Expand / Collapse All"

Analysis with Roboocyte2+ 93 Data List: "Settings" tabbed Page The "Settings" tab includes the following information, from left t

Roboocyte2 Manual 94 Data List "Liquids" tabbed Page In the "Liquids" tab all information about the composition of the liquids a

Analysis with Roboocyte2+ 95 Data List "Injection" tabbed Page The "Injection" tab includes information about the injection of th