Eppendorf Multiporator User Manual
Multiporator
Bedienungsanleitung
Operating Manual
Mode d'emploi
Istruzioni d'impiego
Manual de Instrucciones
Bedienungsanleitung. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4
Operating Manual . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 46
Mode d’emploi. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 89
Istruzioni d'impiego . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 131
Manual de Instrucciones . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 172
EG-Konformitätserklärung. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 213
EC Conformity Declaration
Déclaration de conformité
Dichiarazione di conformità CE
Declaración de conformidad CEE
Nachdruck und Vervielfältigung – auch auszugsweise – nur mit Genehmigung.
No part of this publication may be reproduced without the prior permission of
the copyright owner.
Toute reproduction, complète ou partielle et quel que soit le proèdè est
interdiete, sauf autorisation expresse de notre part.
Ristampa e riproduzione – anche di estratti – solo con autorizzazione.
Reimpresión y copia – incluso parciales – sólo con autorización.
Copyright
©
2006 by Eppendorf AG, Hamburg
B 4308 900.016-10/0606
3
Contents
1 General information
2 Area of application
3 Safety precautions
4 Device description
4.1 Delivery package . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 52
4.2 Startup . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 53
4.3 Cuvette insert. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 53
4.4 Keypad. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 54
4.5 Keys and key combinations . . . . . . . . . . . . . . . . . . . . . . . . . . 54
4.6 Display . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 57
4.7 Electroporation. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 58
4.7.1 Cuvettes and cuvette insert . . . . . . . . . . . . . . . . . . . . . . . . . 58
4.7.2 Electroporation buffer for eukaryotic cells . . . . . . . . . . . . . . . 59
4.8 Cell fusion (optional) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 60
4.8.1 Micro fusion chamber (Electrode gap width 0.2 mm). . . . . . . 60
4.8.2 Micro fusion chamber (Electrode gap width 0.5 mm). . . . . . . 61
4.8.3 Cleaning the Micro fusion chamber . . . . . . . . . . . . . . . . . . . . 61
4.8.4 Helix fusion chamber . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 62
4.8.5 Filling the Helix fusion chamber . . . . . . . . . . . . . . . . . . . . . . 63
4.8.6 Fusion and cell suspension extraction . . . . . . . . . . . . . . . . . . 64
4.8.7 Cleaning and disinfecting the Helix fusion chamber . . . . . . . 65
4.8.8 Electrofusion buffer for eukaryotic cells . . . . . . . . . . . . . . . . . 65
4.9 Insert (electroporation / electrofusion) for connecting
external electrodes. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 66
4.10 Printer connection / printer (optional). . . . . . . . . . . . . . . . . . . 67
5 Mode of operation
5.1 Mode for electroporation of eukaryotic cells . . . . . . . . . . . . . 71
5.2 Mode for electroporation of bacteria and yeast (optional) . . . 73
5.3 Mode for cell fusion (optional) . . . . . . . . . . . . . . . . . . . . . . . . 75
5.3.1 Function: Cell alignment. . . . . . . . . . . . . . . . . . . . . . . . . . . . . 75
5.3.2 Procedure for cell fusion . . . . . . . . . . . . . . . . . . . . . . . . . . . . 77
6 Error messages
7 Maintenance and servicing
7.1 Disinfection . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 82
7.2 Cleaning. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 82
8 Technical data
9 Ordering information
9a Ordering information for North America
. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 47
. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 48
. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 49
. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 51
. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 71
. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 80
. . . . . . . . . . . . . . . . . . . . . . . . . 82
. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 83
. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 85
. . . . . . . . . . . . . . 87
46
1 General information
The Multiporator
With the aid of optional modules, it is also possible to perform
electroporation on bacteria and yeast as well as fusion of cells. This
manual contains information on all possible uses of the Multiporator
The outstanding feature of the Multiporator is that the parameters
entered for the electroporation of eukaryotic cells and for cell fusion are
maintained exactly by means of internal calculation of the discharge
curve.
is designed for the electroporation of eukaryotic cells.
.
47
2 Area of application
Device safety regulations and industrial standards stipulate the following
conditions:
Use the device for experiments in the field of cell technology
–
only
The range of application covers the electroporation of yeast and
bacteria and eukaryotic cells as well as cell fusion.
–
Please use only original accessories from Eppendorf
Original accessories only may be used. They are designed
specifically to ensure optimum functioning of the Multiporator
Additional devices may be used only if recommended by the
manufacturer. Eppendorf does not honor any warranty or accept
any responsibility for damage resulting from the use of incorrect
or non-recommended equipment.
– The device may only be used for research purposes, not for
medical and diagnostic applications
.
48
3 Safety precautions
For reasons of personal safety, it is essential to observe the following!
●
Observe the instructions contained in the operating manual
It is essential to comply with the information given in the manual!
● Only use the device with a power supply which has a grounding
contact!
● Before starting up the Multiporator
supply used corresponds to the voltage specifications on the
identification plate!
● Do not use the device if it is damaged, especially if the mains cable is
damaged.
● The unit may be opened only by authorized service personnel! Before
opening the device, switch it off and remove the power supply plug.
Potentially lethal voltage inside the unit.
● Do not operate the device in an explosive environment!
● Do not allow any liquid to enter into the Multiporator
● The device should be used for in vitro applications only!
● Use the device only for the purpose for which it is intended.
Do not place parts, devices, tools or other objects into the cuvette
insert which have not been recommended by Eppendorf AG.
High voltage!
● Use the listed accessories only.
Other accessories may only be used if they are accompanied by a
safety certificate from Eppendorf AG confirming their suitability for
use!
● Electrical connection to devices which are not mentioned in this
operating manual is only permitted with the prior consent of the
manufacturer!
● The Helix fusion chamber must be handled with care.
Loose or damaged electrodes adversely affect operating procedures.
In such cases, the Helix fusion chamber must not be used!
Do not open the Helix fusion chamber until fusion has finished!
● The Micro fusion chamber must be handled with care.
Ta ke care when filling with cell suspension. Do not touch or move the
electrodes with the pipette tip! This may cause damage.
During cell alignment/fusion, avoid direct contact with the electrodes!
, please ensure that the power
or the inserts!
49
3 Safety precautions
● Cell alignment/fusion may be performed only when the Micro fusion
chamber is connected.
The cable is designed for connection to the Micro fusion chamber only.
No other connections are permitted! Do not place the plug into liquid!
● The owner or operator of the device is liable for the functioning of the
device when the device is serviced or maintained by persons who are
not members of the Eppendorf service personnel or when the device
is not operated in accordance with the given regulations and
precautions. Eppendorf AG accepts no liability for damage resulting
from such action. Warranty and liability stipulations which form part of
the terms of sale and delivery of Eppendorf AG are not affected by the
above-mentioned conditions.
● This operating manual or parts of this operating manual may not be
reproduced in any form without the prior written permission of
Eppendorf AG!
● Eppendorf AG reserves the right to make technical alterations to this
product!
●
Transfer
If the device is passed on to someone else, please include the
instruction manual.
●
Disposal
In case the product is to be disposed of, the relevant legal regulations
are to be observed.
●
Information on the disposal of electrical and electronic devices in
the European Community
The disposal of electrical devices is regulated within the European
Community by national regulations based on EU Directive 2002/96/EC
on waste electrical and electronic equipment (WEEE).
According to these regulations, any devices supplied after 13.08.05 in
the business-to-business sphere, to which this product is assigned,
may no longer be disposed of in municipal or domestic waste. They
are marked with the following symbol to indicate this.
As disposal regulations within the EU may vary from country to
country, please contact your supplier if necessary.
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4 Device description
The device has four different modification levels: the basic device is used
for the electroporation of eukaryotic cells. It can then be developed for the
electroporation of bacteria and yeast, with an additional level offering the
opportunity to perform cell fusion. These two levels can be combined to
form the fourth level.
The Multiporator
The insert for the electroporation cuvette is located next to the keypad.
is operated using an easy-to-follow keypad with display.
Fig. 1: Multiporator, front view
51
4 Device description
4.1 Delivery package
Basic device: Multiporator for the electroporation of eukaryotic
cells; with mains cable
Insert for electroporation cuvettes
Electroporation cuvettes
(Electrode gap width: 2 mm / 4 mm)
Hypo- and isoosmolar buffers for electroporation
Operating manual and Basic Applications Manual
"Bacteria module" optional:
"Cell fusion module" optional:
"Fusion- and bacteria module" optional:
Basic device incl. accessories
with integrated bacteria module
Additional electroporation cuvettes
(Electrode gap width: 1 mm)
Basic device incl. accessories
with integrated cell fusion module
Helix fusion chamber
(Electrode gap width: 0.2 mm)
Insert for Helix fusion chamber
Micro-fusion chamber
(Electrode gap width: 0.2 mm) with special insert
Hypo- and isoosmolar buffers for cell fusion
Basic device incl. accessories
with integrated fusion- and bacteria module.
52
4 Device description
4.2 Startup
The Multiporator must stand completely on the stable work surface and be
safely positioned. There must be enough space that the front and rear
ventilation slits are not covered and that air for cooling can reach
underneath the device.
Space requirement: Width: 25.2 cm
The main power socket with fuse and main power switch is located on the
rear of the Multiporator
tion on the ID plate. To start the device, insert the power cable into the
main power socket and connect it up to the main power supply. Turn on or
off the device by pressing the main power switch.
Depth: 34.2 cm
Height: 12.0 cm
. The mains power must agree with the informa-
Fig. 2: Multiporator, rear view
Power connection (left)
Printer connection (right)
The delivery carton should be kept in order to be able to safely send back
the device in case repairs are necessary.
4.3 Cuvette insert
The insert for the electroporation cuvette is next to the keypad at the
front of the device. To remove the cuvette, pull out the cuvette insert.
When inserting a new cuvette, ensure that the nose of the cuvette fits into
the rear slot. Push the cuvette insert containing the cuvette into the device
up to the stop. A sensor in the device detects whether the cuvette insert
contains a cuvette.
Note:
When transporting the device, please ensure that the insert
contains an empty cuvette. Alternatively, transport the cuvette insert
separately.
Different inserts are used for the modification level "cell fusion".
53
4 Device description
4.4 Keypad
The keypad is used to enter the electroporation or the electrofusion
modes, to select the parameters and to start the application. Depending
on the modification level, 1 to 3 modes can be selected. Key combinations
can be used to set the date and time as well as the volume of the acoustic
signal.
Fig. 3: Control panel with keys and display
4.5 Keys and key combinations
SET
The SET key is used to select the desired parameter. This
parameter is underlined in the display. Further parameters
are selected by pressing the SET key an appropriate
number of times. A SET sequence is ended either by
pressing the SET key repeatedly until the underlining
disappears or by pressing the MODE or START key.
(Note:
The MODE key switches to another mode if
available.) Parameters are modified using the up/down
arrow keys. When the lowest possible value has been
reached, the display then moves to the highest value.
Similarly, when the highest value has been reached, the
display then moves to the lowest value.
54
4 Device description
MODE
The MODE key is used to activate the mode with
which electroporation or electrofusion is to be
performed. It is only possible to select modes in
accordance with the modification level of the device.
This means for the basic device, the mode for the
electroporation of eukaryotic cells is available, while
there is a choice of three modes for the maximum
expansion level.
The corresponding symbol then appears in the
display:
: Mode for eukaryotic cells
Modification level: Basic device
Pulse used: Decaying e-function
: Mode for bacteria and yeasts
Modification level: Bacteria module
Pulse used: Decaying e-function
: Mode for cell fusion
Modification level: Fusion module
Pulse used: Square-wave pulse
START
The process required is started by pressing the
START key.
Arrow keys
The arrow keys are used to modify the selected
(i.e. underlined) parameters in the desired direction.
55
4 Device description
Setting the date
The procedure for setting the date and time can
be activated by pressing the SET and MODE keys
simultaneously. If the SET key is pressed again,
the underlining in the display for the date and time
moves from left to right. The arrow keys can thus
be used to modify the settings.
To end the process for setting the date, press the
SET key repeatedly until the underlining is no longer
visible in the display. Alternatively, the underlining
disappears from the display when the MODE or
START key is pressed.
switches to another mode if available.)
Setting the volume of the acoustic signals
The procedure for setting the volume of the acoustic
signals can be activated by pressing the MODE
and START keys simultaneously. "VOLUME" then
appears in the bottom line of the display, together
with a figure in percent. This figure indicates the
volume and the tone type and can be modified using
the arrow keys.
To end the procedure for setting the volume of the
acoustic signals, press the SET, MODE or START
key.
(Note:
mode if available.) "VOLUME" then disappears from
the display.
(Note:
The MODE key
The MODE key switches to another
56
Reset
An experiment can be terminated only by pressing
the main power switch. This switch also functions as
a reset switch.
4 Device description
4.6 Display
The date and time of day are always displayed in the uppermost line of
the display. The parameters which are relevant for the experiment appear
in lines 2 to 4 in larger letters and in bold print. The parameters and the
number of parameters differ according to the mode which has been
selected. The mode which has been selected is always represented by a
symbol on the right of the bottom line:
: Mode for eukaryotic cells
Modification level: Basic device
: Mode for bacteria and yeasts
Modification level: Bacteria module
: Mode for cell fusion
Modification level: Fusion module
"VOLUME" or the remaining time for the experiment can also appear in
the display in this line.
n
13:37
220µs
1
Example of the display for setting
the volume of the acoustic signal
12.AUG.00
U 500V
VOLUME 100%
τ
During and after the experiment, the display changes to show the
information relevant to the stage of the experiment which has been
reached.
57
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