Multichannel Systems Roboocyte2 User Manual
Roboocyte2 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.
© 2013 Multi Channel Systems MCS GmbH. All rights reserved.
Printed: 17. 06. 2013
Multi Channel Systems
MCS GmbH
Aspenhaustraße 21
72770 Reutlingen
Germany
Fon +49-71 21-90 92 5 - 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.
Table of Contents
Introduction 1
About this Manual 1
Welcome to the Roboocyte2 2
Important Safety Advices 7
Important Safety Advice 7
High Voltage 7
Requirements for the Installation 7
Compressed Air Supply 8
Handling of the Carrier 8
Handling of the Z-Axes 8
Handling of the Ready to Use TEVC Probes 8
Regular Backups 9
Guarantee and Liability 9
Operator's Obligations 9
Setting Up Hardware and Software 11
Setting up Roboocyte2 Hardware and Software 11
Setting up the Roboflow-System 13
Setting Up the Gilson GX-271 Liquid Handler 18
Installing the Software Roboocyte2 23
Testing the Roboocyte2 24
Testing the Movement of Carrier and Z-Axis 24
Testing the Roboflow Valves and Pumps 25
Testing the Amplifier with the Test Model Cell 26
Roboocyte2 Software 33
Introduction 33
Main Window of Roboocyte2 Software 34
Well Plate View - Loading a Well Plate 39
Display Control Functions 41
The Script or Log Window 43
Manual Mode 47
About the Manual Mode 47
Manual Mode Movement 47
Manual Mode Measurement 51
Manual Mode Liquid Handling 56
Compound Application 61
Linking Compound Applications to Recorded Data 61
Using the Roboflow System 65
Using the Gilson Liquid Handler 65
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Roboocyte2 Manual
Recording with Roboocyte2 Control Software 69
Preparations for Recording 69
Alignment 72
Testing the Electrode Resistance 74
Liquid Handling 75
Recording Data 77
Manual Recording Step by Step 77
Automated Recording Step by Step 81
Writing a Script with the built-in Script Editor 82
Analysis with Roboocyte2+ 85
The Roboocyte2+ Analysis Software 85
Analysis with Roboocyte2+ 85
Selecting a Database and Loading a Plate File 88
Analyzer Displays 90
Data Export to ASCII Format 107
Appendix 111
Contact Information 111
iv
1 Introduction
1.1 About this Manual
This manual comprises all important information about the first installation of the Roboocyte2
hardware and software and about the daily work with the Roboocyte2. It is assumed that you
already have a basic understanding of technical and software terms. Thus, no special skills are
required to read this manual.
If you need information on the Roboocyte2 Scripting Language, please refer to the Roboocyte2
Scripting Language manual.
If you are using the Roboocyte2 for the first time, please read the important safety advice before
installing the hardware and software. Please see chapter "First use of the Roboocyte2", where
you will find important information about the installation and first steps.
Then, start practicing using the chapter “Recording in Manual or Automated Mode”. We offer
you the opportunity of "Learning by Doing", which means that you start directly with practicing
without much reading beforehand. We suggest that you start the Roboocyte2 and then follow the
tutorial step by step, either using the integrated help or the printed manual. Just decide what you
like to do, read all necessary information in short and put this information directly into practice.
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” and the “Search” function. 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 Roboocyte2 help. Check also the Multi Channel Systems MCS GmbH web site
www.multichannelsystems.com for downloading up-to-date manuals.
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Roboocyte2 Manual
1.2 Welcome to the Roboocyte2
The Roboocyte2 is a fully-automated all-in-one solution for high-throughput screenings of ligandgated and voltage-gated ion channels, as well as electrogenic transporters based on the standard
Xenopus oocytes expression system.
Main Features
High throughput of experiments.
Recording of 96 oocytes over night.
24 h operation without supervision.
Automated TEVC recording.
Voltage-gated, ligand-activated channels, and electrogenic transporters.
Flexible design of automated recording sequences.
Automated compound application.
Automated cell wash.
2
Introduction
Operation Summary
The integrated TEVC amplifier operates in voltage and current clamp mode.
Instead of having a vast amount of different pull down menus, there is one clearly structured
menu bar and also a toolbar presenting commonly used commands. Therefore, you will quickly
become familiar with the Roboocyte2.
The first step is to set up your "virtual" well plate according to your "real" well plate and the
experiment. The virtual well plate is visualized in a 12 x 8 grid as the real one, and therefore
it is easy to overview and to handle.
For managing high throughput compound screens, enter compounds and concentrations
in the Liquid Handling spreadsheet. This information is linked to the corresponding data traces
and filed into the database.
Select the oocytes that you want to record or load a customizable file.
Then simply start the recording sequence by mouse click. The run will proceed automatically
until finished or interrupted by the user.
Information about the plate and the current state of all oocytes is displayed and updated online
on the screen.
You may save the well plate and load it later to proceed with your experiment or to review
and analyze your data.
Basic analysis features and export filters are included, for example P/n leak subtraction, graphs.
The Roboocyte2 Hardware
The Roboocyte2 is compatible with standard lab equipment and can be easily integrated in your
working environment. Software controls for adjusting the amplifier replace any knobs on the
device. The Roboocyte2 is straightforward and easy to operate; handling does not require special
skills or special equipment.
Recording and cultivation of Xenopus oocytes is performed using disposable standard 96 well
plates, which are commercially available from several providers. MCS recommends 96 well plates
from NUNC. The oocytes are plated into the wells in a couple of minutes and can be kept for
several days. They quickly settle within the cone-shaped wells and adhere to the well bottom
after a few hours. The oocytes do not have to leave the plate anymore; you can easily transfer
the oocytes from the incubator to the Roboocyte2 and back again.
The well plate carrier, powered by linear motors, hovers smoothly and noise free on a cushion
of compressed air above the magnetic x/y table. The carrier operates at a resolution of 20 μm.
The complete system does not require maintenance other than occasional cleaning of the steel
plate.
The vertically moving z-arm, holding the TEVC probe, is designed specifically for the demand
of high speed and precision. The z-arm moves at a resolution of 10 μm; position and speed are
computer-controlled.
The status display of the Roboocyte2 shows the current state of the Roboocyte2 and the
settings.
A quick alignment process guarantees that the oocytes are impaled precisely.
The ClampAmpC is a specifically designed digital TEVC amplifier and is completely automated.
Ready to use TEVC probes allow a quick and easy handling. The ClampAmpC is an advanced
version for the recording of fast sodium channels, with a
faster rise time and a higher
sampling rate.
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Roboocyte2 Manual
The integrated pinch valve Roboflow-System is an ideal perfusion system for drug receptor
characterizations and quick expression tests. The Roboflow is computer controlled and works
virtually pulsation-free, thus minimizing noise during recording.
For high-throughput screens, the Roboocyte2 uses the advantages of an industry-standard liquid
handler from Gilson (optional). The Gilson liquid handler version GX-271 is supported.
The use of the GX-271 liquid handler is an optional feature of the Roboocyte2. Test compounds
are loaded onto the trays and aspirated by the Gilson probe. The liquid is transferred to the
recording well via a peristaltic pump. A transfer port gives access to one or two larger medium
bottles, for example for frog Ringer's or an often used reference compound.
4
Introduction
The Roboocyte2 Software
Operate the Roboocyte2, collect and evaluate the data by using the Roboocyte2 software.
The easy-to-use graphical user interface of the Roboocyte2 software makes daily work with the
Roboocyte2 quick and easy. Recording is started by a single mouse click. The Roboocyte2 controls
the run for all 96 oocytes automatically, even including a wash cycle. Thus the recording can go on
overnight, unsupervised.
You can define different experimental setups not only for different well plates, but also for
specific selections of wells on the same plate. A P/n leak subtraction and several control features
to save time and compounds are provided. Start a wash cycle after a preset time period to
maintain a high viability of the cells.
Automated analysis features are included in the Roboocyte2 software, but you can export the
data to your custom evaluation software as well. You can graph the data and generate reports
with the Roboocyte2 software. The large amount of data generated by the Roboocyte2 can be
managed with a Microsoft Access 2010 database (not included). The database can be accessed
over a network by multiple users.
How is this high degree of automation and flexibility for such a demanding task achieved?
This is possible by using scripts, small text files containing commands. Users can write their
own scripts with any basic text editor or with the editor embedded in the Roboocyte2 program.
All experimental settings are defined within the script. You can write a script for any kind of
experimental setup. Once the appropriate script is loaded into the Roboocyte2 software, simply
click the Run button to start the robot. The script is then performed without the need for further
customization and supervision.
The Java Scripting Language has been specifically designed for the Roboocyte2. It allows the
full automation of various applications. For further information on the Java Scripting Language,
please refer to the separate Java Scripting Language manual or to the help.
5
2 Important Safety Advices
2.1 Important Safety Advice
Warning: Make sure to read the following advice prior to installations of the Roboocyte2.
If you do not fulfill all requirements stated below, this may lead to malfunctions, breakage,
or even fatal injuries. Obey always the rules of local regulations and laws. Only qualified
personnel should be allowed to perform laboratory work. Work according to good laboratory
practice to obtain best results and to minimize risks.
The product has been built to the state of the art and in accordance with recognized safety
engineering rules. The device may only
be used for its intended purpose;
be used when in a perfect condition.
Improper use could lead to serious, even fatal injuries to the user or third parties and damage
to the device itself or other material damage.
Malfunctions which could impair safety should be rectified immediately.
2.1.1 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.
Liquids may cause short circuits or other damage. Keep the power supply and the power cords
always dry. Do not handle it with wet hands.
2.1.2 Requirements for the Installation
The Roboocyte2 weighs more than 22 kg. Always grip it tightly and do not carry it alone,
but with the aid of another person.
The movement of the well plate carrier can lead to vibrations of the workbench on which the
Roboocyte2 is set up. Therefore, the Roboocyte2 must be set up on a rigid, vibration-free base.
The base must also be sufficiently solid to carry the weight of the device.
The Roboocyte2 should be operated only in an air conditioned room. A room temperature
of 20 °C (or less) is recommended. Make sure that the device is not subject to direct sunlight.
It may overheat.
If the air cannot circulate freely around the external power supply, the device may overheat.
Do not shield the power supply by laying anything on top of it.
The external power supply is only for use with the Roboocyte2. Do not connect it to any other
instrument.
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Roboocyte2 Manual
2.1.3 Compressed Air Supply
Even small amounts of water in the compressed air can lead to a corrosion of the carrier. Other
contaminations can also result in malfunctions. Use only compressed air fulfilling the following
standards.
The air must be absolutely free of water, oil, and any particles.
When air is compressed, the humidity in the air is concentrated into a smaller area, frequently
condensing inside the air hoses. Use a water separator to remove water from the air. The
provided water separator is not suitable to establish the necessary conditions; it serves only
as a control device. If you regularly observe an accumulation of water in the provided water
separator, the compressed air does not fulfill the required standards. This is extremely important!
Water in the system leads to corrosion and can destroy the device.
Use a filter with 5 μm pores or smaller to remove any particles from the air.
Set the external air pressure at least to 3.5 bars, the maximum pressure of 5 bars must
not be exceeded. The actual operating pressure should be set to 3 bars by the included pressure
regulator. The Roboocyte2 needs a minimum flow rate of 20 l/min, when the carrier is moving.
Do not use other compressed air hoses than those provided, that is, ID 4 mm OD 6 mm from
compressor to water separator, and ID 2.5 OD 4 mm from water separator to the Roboocyte2.
The use of other hoses could lead to a decrease of pressure.
2.1.4 Handling of the Carrier
Do not try to lift the carrier off the x/y table. You may move the carrier carefully sideways
by hand if necessary (for cleaning purposes, for example), but not during operation.
If it happens that any liquid spills over the x/y table, please remove it as soon as possible in order
to prevent a corrosion of the carrier.
2.1.5 Handling of the Z-Axes
Do not move the z-axes by hand. Breakage may occur. Always use the software controls
to move an axis up and down.
Change the probe with great care. Stabilize the z-axis with one hand while changing the probe
with the other.
The capillaries of the probe are sharp and may lead to injuries. Stay at a safe distance during
operation and protect your eyes. Especially take care not to move your hands in the range of
the z-axes.
Do not try to plug anything other than 0.4 mm wire or the provided connectors into the
sockets of the recording axis. Damage may occur.
2.1.6 Handling of the Ready to Use TEVC Probes
The chloride coated silver wire is sensitive to light. Always keep it dark. Make sure that all
electrodes (including the reference electrodes) are still well-chlorided before you start a recording.
They should look dark grey, not shiny. The Ag/AgCl layer deteriorates over time, leading to a DC
offset and a voltage drift over time.
We recommend that you use the provided connectors to connect the electrodes to the z-axis.
If you want to plug the silver wire directly into the sockets of the z-axis, use only 0.4 mm silver
wire for the electrodes. A wire with a greater diameter will damage the connectors of the z-axis
irreversibly.
8
2.1.7 Regular Backups
You (or the administrator) should perform backups of the Roboocyte2 data files (*.rcd files)
and of the Microsoft Access database (*.accdb) at regular intervals and to appropriate media
for preventing data loss. Data loss may be caused by power failure, system and software errors.
INI file Modifications
If you remove or edit text of an INI file, the software may cause severe problems. Some INI files
relate to hardware functions. A modification of INI files may lead to malfunctions or even severe
damage of the hardware. Always keep a copy of the original INI file. Only advanced users should
modify program files. This warning message applies to all INI file modifications.
2.2 Guarantee and Liability
The General conditions of sale and delivery of Multi Channel Systems MCS GmbH always apply.
The operator will receive these no later than on conclusion of the contract.
Multi Channel Systems MCS GmbH makes no guarantee as to the accuracy of any and all tests
and data generated by the use of the device or the software. It is up to the users to use good
laboratory practice to establish the validity of their findings.
Important Safety Advices
Guarantee and liability claims in the event of injury or material damage are excluded when
they are the result of one of the following.
Improper use of the device.
Improper installation, commissioning, operation or maintenance of the device.
Operating the device when the safety and protective devices are defective and / or inoperable.
Non-observance of the instructions in the manual with regard to transport, storage, installation,
commissioning, operation or maintenance of the device.
Unauthorized structural alterations to the device.
Unauthorized modifications to the system settings.
Inadequate monitoring of device components subject to wear.
Improperly executed and unauthorized repairs.
Unauthorized opening of the device or its components.
Catastrophic events due to the effect of foreign bodies or acts of God.
2.3 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.
9
3 Setting Up Hardware and Software
3.1 Setting up Roboocyte2 Hardware and Software
Connect all cables as described below.
Warning: Carefully lay and secure the cords. Remember that someone could easily trip over
a loose cable.
Note: All electrical connections are clearly marked and the plug coding prevents confusion.
The cords should be plugged in without the use of excessive force
The following photo shows the rear panel of the Roboocyte2
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Roboocyte2 Manual
Connecting the Carrier
1. Plug the carrier connector into the according socket located on the Roboocyte2's rear panel.
2. Connect the Compressed Air Outlet to the carrier.
Warning: Confusion of the compressed air's inlet and outlet may destroy the device. Have
a close look at the preceding picture of the Roboocyte2's rear panel and take care to connect
the lines properly.
Connecting the Roboflow
1. Plug the female connector into the male socket of the Roboflow.
2. Then plug the Roboflow connector into the socket on the Roboocyte2's back.
Connecting the Roboocyte2 to the Pressure Line
1. Connect one end of the provided compressed air hose (ID 2.5 mm, OD 4 mm) to the water
separator.
2. Connect the other end to the compressed air inlet on the Roboocyte2's rear panel.
3. Connect the other provided compressed air hose (ID 4 mm, OD 6 mm) to the provided water
separator.
4. Make sure that the pressure line is closed. Connect the free end of the provided compressed air
hose (ID 4 mm, OD 6 mm) to the pressure line.
5. Open the pressure line and apply a pressure of 3 to 3.5 bar.
Warning: Make sure that the compressed air used fulfills all requirements for working with
the Roboocyte2. Otherwise, use of an improper compressed air could result in physical harm
to the hardware. Especially, check that the compressed air is absolutely free of water, oil,
and any particles. Refer to the chapter "Important Safety Advice" for more information.
Connecting the Roboocyte2 to the Power Line
Finally, plug the power cord into the AC power line input. The diameter of the connector
for the power supply is not round. Please turn the flat part of the connector to the left when
looking directly on the back side of the device. Otherwise the connector will be damaged.
Warning: If the air cannot circulate freely around the external power supply, the device may
overheat. Do not shield the power supply by laying anything on top of it. Make sure it is not
exposed to direct sunlight.
12
3.2 Setting up the Roboflow-System
The Roboflow-System has been especially designed to work with Roboocyte2. The system consists
of two peristaltic pumps and twelve pinch valves. The valve pump for the compounds on the
movable slide of the Roboflow manages the fluidic inflow; the waste pump installed on the right
side of the front panel is for aspiration of the fluids from the wells.
Setting Up Hardware and Software
Assembling the Tubing
The measuring head of the Roboocyte2 carries an inlet and an outlet port for attaching the
perfusion tubing. The liquid is aspirated by the computer-controlled aspiration pump and then
collected in the waste bottle.
Important: Make sure that you do not place any AC voltage sources, that is, any electrical devices
or cables in the immediate vicinity of the Roboocyte2 (especially not near the amplifier or near
any parts belonging to the perfusion system, for example, the bath reservoir or the waste bottle),
as they can introduce a 50 Hz noise or other electrical interferences to your recordings. Move
electrical devices or cables a few inches away if you observe any problems.
The pinch valve system is easy to maintain because the liquid is only in contact with the tubing
material and not with the valves. Simply exchange the tubing in case of contamination. You
need silicone tubing (ID 1.2 mm, OD 2.0 mm) for the pinch valves. One set of preinstalled tubings
is already provided together with the manifold for a quick start.
Please note that materials other than silicone are not flexible enough to seal tightly. If you need
another material for your application though, you can use the silicone tubing for the short way
inside the valve only and connect it to your custom tubing. Use appropriate fittings, which fit
tightly. Also regard the tubing dimensions specified and do not use a thicker or thinner tubing,
otherwise the valves may not work properly or may even be damaged.
Attach the tubing carefully without bending the silicone. Please guide the tubes on there way
from the manifold to the valves through the plastic tube holder to secure the correct attachment.
Do not insert the tubes to close, they need scope for moving with the slide forward and backward.
Please control the provided manifold, specially the connections of the provided tubes to the
manifold.
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Roboocyte2 Manual
Attaching the Manifold
Please control the provided manifold, specially the connections of the provided tubes to the
manifold. If you are going to use only some of the twelve tubes, please connect the useless
tubes to Ringer solution, for example, but do not leave the manifold connections open.
The manifold is fixed via magnetic forces to the housing of the slide of the Roboflow. Adjust the
manifold in a deviation of about 15 degrees to minimize the distance to the tube connection.
Attaching the Tubing to the Valves
Twelve valves regulate the compound flow. Only one valve may be open at a time. Open a valve
by hand or by a software command.
Take one end of tube which is connected to the manifold and insert it gently through the slot on
the right side of the valve. Press the valve with your fingertip to open it manually. Do not insert
the tube to close, it needs scope for moving with the slider forward and backward. Use the plastic
tube guides to secure the tubings inside of the valve slits. Proceed in this manner for all twelve
valves. If you are going to use only some of the tubes, please connect the useless tubes to Ringer
solution, for example, but do not leave single manifold connections open.
14
Setting Up Hardware and Software
Attaching the Tubing to the Valve Pump
Open the screw of the press bracket of the valve pump and remove the press bracket.
Insert the tube section (PPRT1.14) provided for the valve pump.
Remount the press bracket and close the screw carefully. Please start the valve pump by software
control to move the tube section into the pump housing, until the spacer stops this process.
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Roboocyte2 Manual
Software Control for Inserting the Tube Section (PPRT1.14( in the Valve Pump
Please switch to the "Manual Mode" in Roboocyte2 control software. Click the icon
or select main menu "Tools" "Manual Mode". Open the "Liquid Handling" tab page.
Press the button "Valve Pump" to run the pump as long as it needs to insert the tube section into
the pump housing, until the spacer stops this process. Stop the pump by clicking the button again.
Adjust the orientation of the manifold for connecting the short part of tube section to the outlet
nozzle of the manifold. Please see the picture above.
16
Setting Up Hardware and Software
Attaching the Tubing of the Waste Pump
Connect the tubing to the provided connectors of the waste pump (PPRT2.29). The silicone tube
(ID 1.0 mm, OD 2.4 mm, length 540 mm) coming from the left side of the aspiration pump is
directly connected to the outlet of the measuring head of the Roboocyte2. Please guide this tube
also through the plastic tube holder to ensure the correct attachment. The PVC tube (ID 2.0 mm,
OD 4.0 mm, length 1 m) coming from the right side of the aspiration pump leads to the waste
bottle.
Attaching the Tubes to the Measuring Head of the Roboocyte2
Connect the tube section from the valve pump to the inlet connector of the measuring head of
the Roboocyte2. This is the upper one of both provided connections. Connect the suction tube
coming from the waste pump to the outlet of the measuring head which is the lower connection
of the measuring head below the inlet. Please see the picture.
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Roboocyte2 Manual
3.3 Setting Up the Gilson GX-271 Liquid Handler
For high-throughput screens, the Roboocyte2 can be used with an industry standard liquid
handler from Gilson (optional), the GX-271.
Warning: Only hardware configurations and probes provided by Multi Channel Systems
GmbH are supported. Other versions may lead to incompatibilities, up to damages to the
hardware (needle breakage for example). Please verify the system configuration with
Multi Channel Systems GmbH if you obtained the product from a different source other
than Multi Channel Systems GmbH.
Installing the Transfer Port on the Locator Pane
Please see also chapter "Using the Transfer Port".
Important: Please note that only the leftmost rinse, drain, and transfer ports can be used with
the Gilson GX-271 liquid handler. The other transfer ports have to be closed with plugs, you
can use convenient screws. Do not remove the plugs; otherwise, the suction will not suffice
to aspirate solvents from the transfer ports.
1. Connect the provided waste tubing to the waste outlet on the back of the rinse / transfer port
station.
2. Remove the two Phillips screws from the back of the rinse / transfer port station.
3. Raise the cover on the rinse station and align the holes in the rinse / transfer port station with
the holes on the top right side of the locator pan, please see figure "Gilson GX-271 Solvent Deck",
which shows how the Gilson should look like after the installation. (The cover does not need to be
removed completely.)
4. Locate the two Phillips screws included with the rinse station and place them through the holes
on the base of the rinse station, see figure. Tighten the screws using a Phillips screwdriver.
5. Lower the cover on the rinse station and replace the two screws on the back.
18
General Setup of the GX-271 Liquid Handler
Setting Up Hardware and Software
1. Unpack and set up the Minipuls peristaltic pump (MP3) and the Gilson GX-271 liquid handler
according to the descriptions in the Minipuls 3 Peristaltic Pump User’s Guide and the GX-271
Liquid Handlers User's Guide from Gilson in the immediate vicinity of the Roboocyte2 (preferably
on the right side, next to the recording z-arm). Do not place the bath reservoir or the waste bottle
too near to any AC voltage sources, for example the liquid handler itself or the Minipuls pump.
Electrostatic interference can be introduced by the voltage source and travel along the perfusion
line, resulting in a high noise level. Multi Channel Systems recommends that the Minipuls 3 pump
and the Gilson liquid handler are placed on the right-hand side next to the Roboocyte2, please
see the figure "Setup Gilson Liquid Handler", to minimize the travel distance of the solvent.
2. Make sure the rinse / transfer port station was installed to the locator pan. Place the locator
pan onto the locator plate. The rinse / transfer port station should be on the top right position.
The four metal noses of the locator plate should fit into the holes of the locator pane.
3. Set up the z-arm and guide foot according to the chapters "Z-Arm Setup" of the GX-271 Liquid
Handlers User's Guide from Gilson. You will not need the "Fraction Collection Valve Installation".
Adjusting the Z-Arm Height of the GX-271 Liquid Handler
After z-arm installation according to the GX-271 user's manual, you need to adjust the z-arm
height to the appropriate needle length. Please see also chapter "Adjusting the z-travel height"
in the GX-271 user's manual.
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Roboocyte2 Manual
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 illustration.
2. Insert the stop pin into the hole labeled S2, marked by a blue arrow in the preceding illustration.
Installing the Probe and Transfer Tubing
1. Install the provided needle to the GX-271 as described in the chapter "Probe Installation"
of the GX-271 Liquid Handlers User's Guide from Gilson.
2. Install an appropriate pump tubing to the Minipuls 3 pump as instructed in the Minipuls 3
Peristaltic Pump User's Guide from Gilson. Multi Channel Systems recommends the use of
a Tygon pump tubing yellow/yellow, see "Sources of Supply" for details.
3. Screw the FEP tubing assembly onto the probe holder of the GX-271. Connect the other end
of the tubing to the pump tubing by using elastic tubing adapter 200-51 and a 200-16 coupling.
4. Connect the other end of the pump tubing to the Roboocyte2 measuring head via the provided
tubing assembly.
This schematic illustration shows the general setup and the tubing connections of the complete
Roboocyte2 and Gilson liquid handler configuration. The solvent flow (from Gilson tube to
Roboocyte2 recording well) is indicated by the blue arrows.
20
Setting Up Hardware and Software
Electrical Connections
Please note that the Roboocyte2 needs to be connected to the Gilson GX-271 via the Gilson 508
Interface Module.
1. Connect the Gilson z-arm to the z-arm port of the GX-271.
2. Connect the two solenoid valves of the transfer port station to the solenoid valve ports. Make
sure that the cable labeled 1 is connected to the left, and the cable 2 connected to the right port,
as shown in the preceding illustration.
3. Connect the RS232 port of the Gilson 508 Interface Module to the Roboocyte2's serial port
with the provided standard serial cable (see "Connecting the Roboocyte2").
4. Connect the GSIOC port of the Gilson 508 Interface Module to the GSIOC socket of the Gilson
liquid handler with the provided GSIOC cable.
5. Connect the Minipuls peristaltic pump (MP3) to the (same) GSIOC socket of the Gilson liquid
handler with the provided GSIOC cables.
6. Connect the Minipuls peristaltic pump to the power line with the appropriate power cable.
7. Connect the power socket of the Gilson 508 Interface Module to the power line via the provided
external power supply.
8. Connect the power socket of the Gilson liquid handler to the power line with the appropriate
power cable.
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Enabling the Gilson Liquid Handler GX-271 in the Roboocyte2 Program
After installing the Roboocyte2 program, enable liquid handler in the main menu "Settings",
"Options" dialog box. After changing the settings in the check box, please close the Roboocyte2
software and start the program again to activate the communication with the Gilson.
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3.4 Installing the Software Roboocyte2
The operating system Microsoft Windows ® 7 (32 and 64 bit) or XP (with Service Pack 3) and the
Roboocyte2 software are already installed on the provided computer. However, you may need
to reinstall or update the Roboocyte2 software on the same or another computer. Please check
the system requirements if you are going to install the software on another than the provided
computer. MCS cannot guarantee that the software works properly if these requirements are
not fulfilled.
If you want to access the same data files on a network server from a second computer (for offline
analysis), you have to manually relink the original database from the second computer after the
program installation. A new database will be set up during program installation that you will not
need. Please see chapter "Linking to the Database" for more information.
Otherwise, it is possible that the installed software does not work properly.
Double-click Setup.exe on the installation volume.
The installation assistant will show up and guide you through the installation procedure.
Setting Up Hardware and Software
Follow the instructions of the installation assistant until the installation is complete.
The Roboocyte2 software and the Microsoft Access database file (*.accdb) are installed
on your computer.
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3.5 Testing the Roboocyte2
Each delivered Roboocyte2-System has undergone extensive tests at the MCS site. Please follow
the provided instructions to perform initial tests just to make sure that everything is alright and
that no damage has occurred during shipment.
3.5.1 Testing the Movement of Carrier and Z-Axis
On the main menu "Tools" click "Manual Mode" or click the icon . The manual mode dialog
box opens with the "Movement" tab page active.
Warning: DO NOT press the "Set Alignment" button. This button should only be touched
with installed measuring head aligned to the surface of the alignment tool.
The "Movement" tab is divided into different sections: "Global, Plate Carrier, Air Pressure,
Resistance, and Z-Axis". For a detailed description of all functions please refer to chapter
"Manual Mode"
Testing the Air Pressure
Press the "Check" button in the Air Pressure section. The actual air pressure will be displayed.
Testing the Reference Movement
Press the "Reference" button in the "Global" section. The z-axis and the well plate carrier
should perform their reference movements.
Getting familiar with the Movement Tab Page
Try the other buttons of the "Movement" tab window - you will quickly find out what they do.
Important again: DO NOT press the "Set Alignment" button. This button should only be pressed
with installed measuring head aligned to the surface of the alignment tool.
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3.5.2 Testing the Roboflow Valves and Pumps
The Roboflow was designed to work with the Roboocyte2 and is therefore completely controlled
by the Roboocyte2 software.
For an initial test, open the "Liquid Handling" tab of the "Manual Mode" window.
Setting Up Hardware and Software
Testing the Pinch Valves
Press the valve buttons one after the other from 1 to 12 and check whether the respective valves
open.
Testing the Peristaltic Pumps
Press the "Valve Pump" button and check whether the pump located on the movable slide runs.
Then press the "Waste Pump" button and check whether the pump on the upper right side of the
Roboflow front panel runs. The number of the pump "Speed" is roughly equivalent to μl/min, the
maximum speed for pumps is 10000 and 20000 for the valve pump and waste pump, respectively.
Overall Test
You can also use the "Start" button in the "Clean Valves" section; this will open all valves selected
between "First Valve" and "Last Valve" for the specified "Rinse Time". At the same time both
peristaltic pumps will be started at the speed selected. After finishing the cycle all valves will be
closed, the valve pump will stop and the waste pump will continue to operate. If you want to stop
it, you just have to press the Waste Pump button again.
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3.5.3 Testing the Amplifier with the Test Model Cell
You can use the provided test model cell (TMC) to test the integrated digital TEVC amplifier.
For initial testing, you can measure the resistance and run a voltage step protocol. This is also
a good way to make your first experience with the recording features. The TMC mimics the
electrical (passive) properties of a real oocyte impaled with microelectrodes (electrode resistance
~450 kOhm, membrane resistance 100 kOhm, membrane capacitance ~ 200 nF).
Connecting the Test Model Cell
1. Plug the current electrode connector (red) and the corresponding reference (black) of the test
model cell into the sockets on the left side of the z-axis (red to front, black to back).
2. Plug the voltage electrode connector (blue) and the corresponding reference (black) of the
test model cell into the sockets on the right side of the z-axis (blue to front, black to back).
Measuring the Electrode Impedance of the Test Model Cell
On the "Movement" tab of the "Manual mode", click button "Check" in the "Resistance"
window.
The electrode resistances for current and voltage electrode are displayed in the two small
windows beside. Both impedances should be approximately 500 kilohms.
Performing Voltage-Clamp with the TMC
You can run a test recording with the test model cell in order to check the speed and accuracy of
the voltage-clamp either in continuous recording mode or by a applying a voltage-step protocol.
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