Multichannel Systems Roboinject User Manual
Roboinject 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: 19. 07. 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
Important Information and Instructions 3
Important Safety Advice 3
High Voltage 3
Requirements for the Installation 3
Compressed Air Supply 4
Handling of the Carrier 4
Handling of the Injector Mount ( = z-Axis) 4
Handling of Injection Needles 4
Regular Backups 4
System File Modifications 5
Guarantee and Liability 5
Operator's Obligations 5
Roboinject 7
Welcome to the Roboinject 7
First Use of the Roboinject 9
Installing the Roboinject 12
Installing the Roboinject Software 14
Roboinject Software 15
Main Menu 16
Toolbar 29
Control View 35
Tube View 40
Well Plate View 42
Injection 47
About Injection with the Roboinject 47
Strategies for a Successful Injection 48
Preparations for Injection 50
Setting Up the Well Plate 52
Inserting or Changing the Well Plate 52
Selecting Oocytes for Injection 57
Changing the Well Plate and Loading the Needle 58
Alignment 60
Starting the Injection 63
Saving the Well Plate 65
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Roboinject Manual
Troubleshooting 67
Troubleshooting 67
Clogged Injection Needle 68
Error Messages 70
Appendix 71
Technical Support 71
Service and Maintenance 72
Cleaning the x/y Table 72
Replacing the Plunger 72
Contact Information 73
Ordering Information 74
About Preparation of Xenopus Oocytes 76
Materials 76
Chemicals 77
Oocyte Removal 78
Collagenase Digestion and Defolliculation 78
Selecting Good Oocytes 79
Plating Oocytes 80
Technical Specifications 81
Index 83
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1 Introduction
1.1 About this Manual
This manual comprises all important information about the first installation of the Roboinject
hardware and software and about the daily work with the Roboinject. 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 are using the Roboinject for the first time, please read the "Important Safety Advice"
before installing the hardware and software. Please see chapter "First Use of the Roboinject",
where you will find important information about the installation of hardware and software.
The printed manual and online 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 Roboinject help. Check also the Multi Channel Systems MCS GmbH web site
(www.multichannelsystems.com) for downloading up-to-date manuals.
1
2 Important Information and Instructions
2.1 Important Safety Advice
Warning: Make sure to read the following advice prior to installations of the Roboinject.
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 Roboinject weighs more than 23 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
Roboinject is set up. Therefore, the Roboinject 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 Roboinject 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 unit is only for use with the Roboinject. Do not connect it to any
other instrument.
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Roboinject 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.
Warning: Water in the system leads to corrosion and destroys the device!
This is extremely important!
Use a filter with 5 μm pores or smaller to remove any particles from the air.
Set the external air pressure at least to 5 bar, the maximum pressure of 10 bar must not be
exceeded. Adjust the provided pressure regulator to 3 bar, too high or too low pressure can
lead to malfunction and can severely damage the device. Check the pressure at the end of the
pressure line, directly before it is lead into the Roboinject. The Roboinject needs a minimum
flow rate of 20 l/min.
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 Roboinject.
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 but not during operation. For cleaning or alignment purposes use the
appropriate software control, please read chapter "Maintenance" and "Alignment".
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 Injector Mount ( = z-Axis)
Do not move the z-axis by hand. Breakage may occur. Always use the software controls
to move an axis up and down, please see chapter “Manual Mode".
Change the injection needle with great care. The injection needle is 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-axis.
2.1.6 Handling of Injection Needles
The injection needles are sharp and break easily. Always handle them with care.
In rare cases, a needle may splinter when inserting the plunger into the needle and fitting it.
Stay at a safe distance and protect your eyes.
Dispose the used injection needles into an appropriate container.
2.1.7 Regular Backups
You (or the administrator) should perform backups of the Roboinject plate files (*.rpf files) and
template files (*.rit files) at regular intervals and to appropriate media for preventing data loss.
Data loss may be caused by power failure, system and software errors.
4
Important Information and Instructions
2.1.8 System File Modifications
If you remove or edit text of an initialization ("ini") or configuration ("cfg") file, the software may
cause severe problems. These files relate to hardware functions. A modification of these files may
lead to malfunctions or even severe damage of the hardware. Always keep a copy of the original
files. Only advanced users should modify program files like these. This warning message applies to
all initialization and configuration 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.
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.
5
3 Roboinject
3.1 Welcome to the Roboinject
The Roboinject is a fully-automated all-in-one solution for injection of cDNA, mRNA and other
compounds into Xenopus oocytes in standard 96 well plates. It is also suited for automated
injection of other cells or organisms in standard (96, 384) as well as customized well plates.
Main features:
Compact and functional design, works fully computer controlled.
Easy handling - neither special skills nor special equipment required.
Maintenance-free system.
Extremely simple and stable mounting of injection capillaries.
Injection is performed by a positive displacement motorized unit which is capable
of repetitively dispensing precisely 1 to 100 nl per cell or organism.
Reproducible and defined cell impalement depth.
Automated sample uptake of up to eight different RNAs, DNAs or compounds
per well plate from industry standard 0.5 ml reaction tubes.
Automated and effective needle rinsing before uptake of new samples.
Injection of up to four different samples per cell.
Full documentation of all injection parameters.
Compatibility with the Roboocyte when using 96 well plates.
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Roboinject Manual
Operation summary:
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 Roboinject.
The first step is to set up your "virtual" well plate according to your "real" well plate.
Enter a unique ID for the plate and / or assign a file name. The virtual well plate is then visualized
in a X x Y grid (for example, 12 x 8 for a standard 96 well plate) as the real one, and therefore
it is easy to overview and to handle.
Define up to eight different injection samples and select the respective velocity of injection.
Enter the respective sample volume.
Finally, assign the different injection samples to a selection of wells, rows or columns. To allow
maximal flexibility, you can inject up to four samples per cell - even on different days - and assign
injection depth and injection volume for every cell independently.
Fill the injection needle with mineral oil and lock it to the injector's luer connector.
Start the injection sequence by mouse click. The "Injection Wizard" will open and guide you
through a quick and easy alignment procedure. Finally, you start the injection and it will proceed
automatically until finished or interrupted by the user.
Information about the plate and the current state of all wells is displayed and updated online on
the screen.
You may save the well plate and load it later on the Roboocyte to proceed with your experiment.
8
3.2 First Use of the Roboinject
This section contains important information about the general concept of the Roboinject and
the setup of the system. Please read the "Important Safety Advice" in the introduction prior to
installing the Roboinject hardware and software.
The Roboinject Hardware
The Roboinject is compatible with standard lab equipment and can be easily integrated
in your working environment. Software controls for all settings replace any knobs on the device.
The Roboinject is straightforward and easy to operate; handling does not require special skills or
special equipment.
Roboinject
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 well plate carrier can hold up to eight industry standard 0.5 ml reaction tubes serving
as sample reservoirs.
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Roboinject Manual
The vertically moving injector mount (z-axis), holding the injection device, is designed
specifically for the demand of high stability, speed and precision. It moves at a resolution
of 20 μm; position and speed are software controlled.
The injection device is composed of a linear motor driving a steel plunger which fits precisely
in the provided injection capillaries. The oil gap between the inner glass wall and the plunger
surface serves for a perfect leak tightness and makes additional gaskets dispensable.
Prepulled borosilicate glass micropipettes are provided by Multi Channel Systems MCS GmbH,
but custom injection needles may also be used. Mounting of injection capillaries is extremely fast
and simple by using luer fittings. There are no additional gaskets susceptible to leakage rendering
the injection device durable and maintenance free.
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Roboinject
Injection and cultivation of Xenopus oocytes is performed using disposable standard 96 well
plates, which are commercially available from several providers. Please see chapter "Ordering
Information" in the Appendix. 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 Roboinject and back again.
A quick alignment process guarantees that the oocytes are injected precisely. The cDNA or mRNA
or other compounds to be injected are aspirated into the injection pipette by upward movement
of the plunger and then pushed into either the nucleus or the cytoplasm by downward movement
of the plunger.
The provided microscope is mandatory for the alignment process. Place the microscope
in a position onto the x/y table which allows to focus on the alignment device plugged
in well H12 of the well plate. Please read chapter “Alignment” for detailed information.
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Roboinject Manual
3.3 Installing the Roboinject
Setting Up the Roboinject
1. Place the Roboinject on a rigid, stable, and vibration-free base in an air conditioned room.
2. Provide a power supply in the immediate vicinity of the installation site.
3. Provide a compressed air supply in the immediate vicinity of the installation site.
4. Place the computer and its accessories next to it.
Connecting the Roboinject
The following illustration shows the rear panel of the Roboinject.
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.
Connecting the carrier
Plug the carrier connector into the according socket located on the Roboinject's rear panel,
labelled with "CARRIER OUT".
Warning: Confusion of the compressed air's inlet and outlet may destroy the device.
Have a close look at the preceding picture of the Roboinject's rear panel and take care
to connect the lines properly. The air hose connecting the Roboinject with the well plate
carrier is close to the electrical connection between them.
12
Roboinject
Connecting the Roboinject 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 Roboinject's rear panel, labelled
with "AIR IN".
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, labelled with "AIR OUT".
5. Open the pressure line and apply a pressure of 3 bar with the provided pressure regulator.
Warning: Make sure that the compressed air used fulfills all requirements for working with
the Roboinject. 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 Roboinject to the computer
1. Connect the Roboinject by the provided USB cable, labelled with "USB".
2. Connect the Roboinject to the power line.
3. Finally, plug the power cord into the AC power line input, labelled with “24 VDC”.
4. Switch the Roboinject on or off via “I/O” toggle switch.
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.
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Roboinject Manual
3.4 Installing the Roboinject Software
System requirements
Software: One of the following Microsoft Windows ® operating systems is required:
Microsoft Windows 7, Vista or XP (English versions supported with the NT file system).
Other language versions may lead to software errors.
Hardware: An USB 2.0 High Speed serial bus must be provided for the communication
between the Roboinject and the computer. Additional 1 GB RAM working memory
is strongly recommended.
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. These routines were
designed for use on office computers, but are not very useful for a data acquisition computer.
Turn off “Windows System Restore”.
Turn off automatic “Windows Update”.
Deselect “Windows Indexing Service” for all local disks.
Turn off “Optimize hard disk when idle” (automatic disk fragmentation).
It is also not recommended to run any applications in the background when using the Roboinject.
Remove all applications from the Autostart folder.
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.
Installing the Roboinject Software with computer connected
Power up the connected computer and wait until it is ready.
1. Double-click Setup.exe on the installation volume. The installation assistant will open
and guide you through the installation procedure.
2. Follow the instructions of the installation assistant. The Roboinject software and all necessary
drivers will be installed on your computer.
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3.5 Roboinject Software
Starting the Roboinject
Press the main switch on the back of the Roboinject for switching on the Roboinject.
The Roboinject is starting now. After a few seconds the Roboinject is ready for use.
Starting the Roboinject Software
You may use the computer and the software offline that is, the Roboinject is switched off or not
connected to the computer, to review already injected plates or define new injection protocols.
You can switch from offline to online mode, when the Roboinject is switched on later. However,
if you start the software to operate the Roboinject, the easiest way is to start the Roboinject first
and then start the software.
Each time you start the Roboinject software the well plate carrier and the injection mount will
automatically move to a reference point, that means to the "home" position. These movements
do not use any software command.
The factory preset default values have been extensively tested with RNA and DNA injection of
Xenopus oocytes and should work in most circumstances. Therefore, it is advisable to use the
default values for the first injections with Xenopus oocytes.
Roboinject
1. Double-click the Roboinject icon
- OR -
Select Roboinject from the "Start" menu to start the Roboinject software.
2. When you start the Roboinject software for the very first time, the file dialog box appears
for setting the directories for the plate files and template files. You can either accept the
predefined directories or select directories according to your personal preferences.
If the Roboinject is offline indicated by a red status lamp below the Roboinject logo,
switch the Roboinject on and wait until the status lamp turns green.
on the desktop.
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Roboinject Manual
The Roboinject main window opens.
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3.6 Main Menu
For maximum flexibility all relevant parameters for the injection can be changed by the user
via main menu.
Note: The preset default values have been extensively tested with RNA and DNA injection of
Xenopus oocytes and should work in most circumstances. Therefore, it is advisable to use the
default values for the first injections with Xenopus oocytes.
File
Roboinject
Menu for creating new files, opening existing files, and for saving them. Menu for opening
and saving templates, and for closing the program.
The option "Export to Roboocyte1" is a feature for exporting plates to the Roboocyte1 device
from Multi Channel Systems MCS GmbH.
Well plate files, created and saved with Roboinject software, can be transferred to the data base
of Roboocyte1 software. The corresponding well plates, injected with the Roboinject device can
be processed with the Roboocyte1 device. The feature is available only for owners of Roboocyte1
devices (Clamp Amplifier A and Clamp Amplifier B) and together with Roboinject software
version 1.1.0 which includes a firmware update. It is not possible to export plates to other devices.
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Roboinject Manual
Click "New" in "File" menu. The following "Plate Info" dialog opens.
Please give an appropriate file name in "Plate ID". The type of the well plate can be defined
from the "Plate Type" drop down menu. If you want to set the selected plate type as default,
please enable the check box "Set As Default". You can type in notes concerning the actual
experiment into the "Comment" text box. All information will be stored in the plate info (*.rpf)
file, and displayed in the report sheet.
If you want that the plate ID is automatically counted up when creating a new file, please enable
the check box “Auto increment plate ID” in “General” tab of the “Options” dialog in main menu
“Settings”.
Click "OK" to enter additional dialogs, or to start the injection.
If you want to inject the same plate again or review previously injected cells, open the appropriate
well plate file.
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Click "Open" in "File" menu. The "Open" dialog box appears.
Roboinject
Browse your folders and select the Roboinject plate info file (*.rpf) you want to open.
Click "Open". The main window now shows the state of the already saved plate.
Click "Save" to save a Roboinject plate info (*.rpf) file. It will be stored in the selected
"Plate Info" directory.
Click "Save Template" and "Open Template" to open and save a template (*.rit) file.
It will be stored in the selected "Template" directory, respectively.
Click "Exit" to close the Roboinject software program.
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Roboinject Manual
Settings
Menu for setting options and for defining directories.
Click "Options" in main menu "Settings". The "Options" dialog has four tabs.
Click "General" tab.
The dialog allows general settings concerning two general software features,
and the "Test Injection Dialog" in particular.
Enable the check box "Auto increment plate ID", if you want that the plate ID
is automatically counted up when creating a new file.
Enable the check box "Reset / Remove Microscope dialog" if you want to see
the “Remove Microscope” dialog after alignment again.
The “Test Injection” dialog appears immediately before the injection run of each sample
if selected in the "Sample Information" dialog. This allows the user to execute a test
injection before starting the injection run or to stop the injection run completely.
With regard to the "Test Injection Dialog" you can select or deselect the two check boxes
"Enable Auto Close" and "Beep before dialog closes". Additionally you can choose
the time span for displaying the "Test Injection Dialog" from the drop down menu.
The "Auto close time" varies from 30 to 180 seconds.
20
Click "Aspirate" tab.
Roboinject
This dialog allows to define the aspiration of compound from the reservoir tubes.
The parameters are the “Velocity” of aspiration in nl/s, the “Surplus Volume” in nl,
and the “Pause after Aspirate” in ms.
“Velocity”: The aspiration velocity for a given compound depends on its viscosity. Samples
with high viscosity should be aspirated slower than samples with low viscosity. This prevents
the accumulation of negative pressure inside of the injection needle leading to the formation
of air bubbles inside of the capillary.
“Surplus Volume”: In order to prevent the injection of minute volumes of mineral oil -
the needle was filled with first - with the last injection, you can define a "Surplus Volume".
The volume will be aspirated additionally to the injection volume, defined in the "Injection" tab.
Values of 50 nl are recommended.
"Pause after Aspirate": You can define a time period during which the injection needle stays
in the compound after aspirating the RNA, DNA or other compounds, before the injector mount
moves upward. This feature minimizes the risk of sucking air into the needle tip. A time span of
2000 ms is recommended.
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Roboinject Manual
Click "Inject" tab.
The dialog allows to define parameters concerning the “Injector Mount” (z-axis) and the
“Injector” device itself. The settings are attributed to the different "Sample Types" such
as RNA, DNA or others.
“Sample Type”: Define the type of compound to which you want to assign the setting
parameters from the drop down menu.
“Injector Mount = z- Axis”:
You can separate the impalement process of the injection needle into an oocyte or another cell
in two phases. This is especially useful, for example, when working with "soft" oocytes. If the
first movement (for example, to 650 μm above well bottom) only pushes down the cell without
penetrating the vitelline layer and membrane, a second movement (for example, to 550 μm
above well bottom) will most likely lead to a successful impalement. Please see the picture.
Please define the parameters of the first step, called “Impalement Position”. Please note that
the value in micrometer refers to the distance between the tip of the needle and the well bottom.
Define the “Velocity” of impalement in mm/s. The default value of 30 mm/s was intensively
tested with oocytes and will work in most of the cases.
Please define the parameters of the second step, called “Injection Position”. Define the position
of the injection in μm from the up-down control. Please note that the value in micrometer refers
to the distance between the tip of the needle and the well bottom in relation to the bottom of
the well. Define the “Velocity” of injection in mm/s. Again, the default of 30 mm/s will work with
most oocytes.
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Roboinject
Example on the picture: The bottom of the well is value zero, the reference value. If you define,
for example an impalement position of 650 μm, the injector mount will move down until its
position is 650 μm above the bottom of the well. If you additionally define an injection position
of 550 μm, the injector mount will move down again until its position is 550 μm above the bottom
of the well.
If you define, for example an impalement position of 300 μm, the injector mount will move down
until its position is 300 μm above the bottom of the well. If you additionally define an injection
position of 550 μm, the injector mount will move up until its position is 550 μm above the bottom
of the well.
The “Wait between Movements” in ms as well as the "Pauses before and after Injection"
can be adjusted to user preferences.
“Injector”
The injection interval is terminated by the time frame before injection "Pause before
Injection", and the time frame after injection "Pause after Injection", both measured in ms.
The default values for the wait times before and after injection (500 ms and 2000 ms, respectively)
have been tested with oocytes and RNA injections.
You have to define the injection “Volume” in nl, and the “Velocity” in nl/s from the up-down
controls. The respective default values (50 nl and 70 nl/s) have been tested with oocytes and
should work in most cases.
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Roboinject Manual
Adjusting the Injection Parameters for each Well separate
It is possible to adjust the settings for the injection for each single well separate. Click with
the right mouse button onto a well in the virtual well plate, and the following dialog appears.
Now you can adjust the injection parameters for this specific well and for each sample separate.
If the oocyte in the specific well should be injected with different samples, you can adjust
the injection parameters for each sample separately. Please click the "Sample" tabs.
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