Titel_d_e_f_i_sp.fm Seite 1 Freitag, 9.
Titel_d_e_f_i_sp.fm Seite 2 Freitag, 9.
Inhalt.fm Seite 3 Freitag, 9. Juni 2006 8:03 08 Bedienungsanleitung. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4 Operating Manual . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 46 Mode d’emploi. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 89 Istruzioni d'impiego . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 131 Manual de Instrucciones . . . . . . . . . . . . . . . . . . .
Inhalt_en.fm Seite 46 Freitag, 9. Juni 2006 8:04 08 Contents 46 1 General information . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 47 2 Area of application. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 48 3 Safety precautions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 49 4 4.1 4.2 4.3 4.4 4.5 4.6 4.7 4.7.1 4.7.2 4.8 4.8.1 4.8.2 4.8.3 4.8.4 4.8.5 4.8.6 4.8.7 4.8.8 4.9 51 52 53 53 54 54 57 58 58 59 60 60 61 61 62 63 64 65 65 4.
Multiporator_Text_en.fm Seite 47 Freitag, 9. Juni 2006 8:05 08 1 General information The Multiporator is designed for the electroporation of eukaryotic cells. 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.
Multiporator_Text_en.fm Seite 48 Freitag, 9. Juni 2006 8:05 08 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.
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Multiporator_Text_en.fm Seite 50 Freitag, 9. Juni 2006 8:05 08 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.
Multiporator_Text_en.fm Seite 51 Freitag, 9. Juni 2006 8:05 08 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 is operated using an easy-to-follow keypad with display.
Multiporator_Text_en.fm Seite 52 Freitag, 9. Juni 2006 8:05 08 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: Basic device incl.
Multiporator_Text_en.fm Seite 53 Freitag, 9. Juni 2006 8:05 08 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 Depth: 34.2 cm Height: 12.0 cm The main power socket with fuse and main power switch is located on the rear of the Multiporator.
Multiporator_Text_en.fm Seite 54 Freitag, 9. Juni 2006 8:05 08 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.
Multiporator_Text_en.fm Seite 55 Freitag, 9. Juni 2006 8:05 08 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.
Multiporator_Text_en.fm Seite 56 Freitag, 9. Juni 2006 8:05 08 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.
Multiporator_Text_en.fm Seite 57 Freitag, 9. Juni 2006 8:05 08 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.
Multiporator_Text_en.fm Seite 58 Freitag, 9. Juni 2006 8:05 08 4 Device description 4.7 Electroporation 4.7.1 Cuvettes and cuvette insert The electroporation of eukaryotic cells, bacteria or yeasts is carried out in disposable cuvettes. The biological material is placed in the gap between the electrodes of the cuvette, whereby the prescribed liquid volumes of the relevant cuvette are to be maintained.
Multiporator_Text_en.fm Seite 59 Freitag, 9. Juni 2006 8:05 08 4 Device description 4.7.2 Electroporation buffer for eukaryotic cells To achieve optimal transfection results, the original electroporation buffer from Eppendorf, with a low electrical conductivity, should be used. These electroporation buffers (hypo and isoosmolar) are tested for sterility and the absence of mycoplasms and endotoxins.
Multiporator_Text_en.fm Seite 60 Freitag, 9. Juni 2006 8:05 08 4 Device description 4.8 Cell fusion (optional) 4.8.1 Micro fusion chamber (Electrode gap width 0.2 mm) With the help of the Micro fusion chamber, the parameters for the cell alignment and for the cell fusion can be optimized under microscopic control. The Micro fusion chamber consists of a housing equipped with a transparent reservoir, into which two electrodes with a gap of 200 µm are embedded.
Multiporator_Text_en.fm Seite 61 Freitag, 9. Juni 2006 8:05 08 4 Device description – Connect the Micro fusion chamber with the coaxial connection of the cable. – Focus the electrodes and cells under the microscope. – Run through the parameters by pressing the START key. This can be monitored under the microscope (see chap. 5.3). 200 µm Alignment of the cells between the electrodes Cells after fusion and fusion products Fig.
Multiporator_Text_en.fm Seite 62 Freitag, 9. Juni 2006 8:05 08 4 Device description 4.8.4 Helix fusion chamber The Helix fusion chamber has been specially designed for the production of larger quantities of fusion products (hybrids). It consists of a conical tapered core that carries the electrodes and a beaker into which the cell suspension is added (see fig. 7). Fig. 7: Helix fusion chamber Diagram of Helix fusion chamber The Helix fusion chamber (see fig.
Multiporator_Text_en.fm Seite 63 Freitag, 9. Juni 2006 8:05 08 4 Device description The gap between the wound platinum electrodes is approx. 200 µm. 200 µm Fig. 9: Microscope view of the electrodes of the Helix fusion chamber Caution: Handle the Helix fusion chamber with care! Loose or damaged electrodes adversely affect operating procedures. In such cases, the Helix fusion chamber must not be used! 4.8.5 Filling the Helix fusion chamber The Helix fusion chamber is filled with 250 µl cell suspension.
Multiporator_Text_en.fm Seite 64 Freitag, 9. Juni 2006 8:05 08 4 Device description When screwing the core, make sure that no air bubbles form. Fig. 10: Filling the Helix fusion chamber (schematic representation) Leave the closed Helix fusion chamber upside-down on the coaxial connector until the fusion process has been completed! 4.8.6 Fusion and cell suspension extraction – Slide the fusion insert firmly into the device.
Multiporator_Text_en.fm Seite 65 Freitag, 9. Juni 2006 8:05 08 4 Device description 4.8.7 Cleaning and disinfecting the Helix fusion chamber The beaker and core of the Helix fusion chamber should be rinsed with distilled water directly after the experiment in order to prevent cell and buffer residue from drying up. If heavily contaminated, the Helix fusion chamber should be cleaned briefly in an ultrasonic bath (possibly with a cleaning supplement, such as Edisonite Super) or with a very soft (tooth)brush.
Multiporator_Text_en.fm Seite 66 Freitag, 9. Juni 2006 8:05 08 4 Device description 4.9 Insert (electroporation / electrofusion) for connecting external electrodes Prior to using the insert for the connection of external electrodes, the supplement sheet accompanying the insert must be read thoroughly. The safety instructions found in the supplement sheet must be observed. The non-standard accessories (see "Ordering information") contain an insert, to which it is possible to connect external electrodes.
Multiporator_Text_en.fm Seite 67 Freitag, 9. Juni 2006 8:05 08 4 Device description When external electrodes are connected, the relevant national safety regulations (relating to high voltage) and EMC regulations apply and should be observed (EMC: electro-magnetic compatibility). The user is responsible for all equipment that has been connected! 4.10 Printer connection / printer (optional) The Multiporator can supply a printer protocol for each experiment.
Multiporator_Text_en.fm Seite 68 Freitag, 9. Juni 2006 8:05 08 4 Device description 18.Aug.00 16:42:47 eppendorf Multiporator 4308 Bacteria and Yeast U = 1000 V, Ua = 992 V, Cuvette Gap Width 1mm 2mm V4.00 τa = 5.
Multiporator_Text_en.fm Seite 69 Freitag, 9. Juni 2006 8:05 08 4 Device description 18.Aug.97 15:59:08 eppendorf Multiporator 4308 V1.00 ******** ERROR 07 ******** ******** Short τ ********* Serial printers can be connected to the serial printer interface at the rear of the device. It is also possible to read protocols for documentation purposes via a PC terminal program. This requires a zero-modem cable (9-pin socket / 9-pin socket, order no. 0013 610.525).
Multiporator_Text_en.fm Seite 70 Freitag, 9. Juni 2006 8:05 08 4 Device description DIP SW settings For Thermal printer DPU-414 e.g.
Multiporator_Text_en.fm Seite 71 Freitag, 9. Juni 2006 8:05 08 5 Mode of operation 5.1 Mode for electroporation of eukaryotic cells This mode is represented in the display by the symbol . Instructions for the filling of the electroporation cuvettes have been summarized in chapter 4.7.1 of the operating instructions. A detailed description of the electroporation of eukaryotic cells can be found in the Basic Applications Manual for electroporation, which is included in the delivery package.
Multiporator_Text_en.fm Seite 72 Freitag, 9. Juni 2006 8:05 08 5 Mode of operation Step 2 – Starting electroporation – Trigger the electroporation process by pressing the START key. – "Charge" appears in 12.AUG.00 13:37 the third line of the U 500V τ 220µs display. Charging Charge commences. – After the charging procedure has ended, discharging occurs. This is indicated by a flash in the third line of the display.
Multiporator_Text_en.fm Seite 73 Freitag, 9. Juni 2006 8:05 08 5 Mode of operation 5.2 Mode for electroporation of bacteria and yeast (optional) This mode is represented in the display by the symbol . Instructions for the filling of the electroporation cuvettes have been summarized in chapter 4.7.1 of the operating instructions. In addition, application protocols for the electroporation of bacteria, yeasts and other microorganisms can be downloaded from the Eppendorf Homepage www.eppendorf.com.
Multiporator_Text_en.fm Seite 74 Freitag, 9. Juni 2006 8:05 08 5 Mode of operation Step 2 – Starting electroporation – Trigger the electroporation process by pressing the START key. – "Charge" appears in 12.AUG.00 13:42 the third line of the U 2000V τ 5 ms display. Charging Charge commences. – After the charging procedure has ended, discharging occurs. This is indicated by a flash in the third line of the display, accompanied by a double acoustic signal. After the experiment has 12.AUG.
Multiporator_Text_en.fm Seite 75 Freitag, 9. Juni 2006 8:05 08 5 Mode of operation 5.3 Mode for cell fusion (optional) This mode is represented in the display by the symbol . Instructions for the usage of the Micro fusion chamber and the Helix fusion chamber have been summarized in chapter 4.8 of the operating instructions.
Multiporator_Text_en.fm Seite 76 Freitag, 9. Juni 2006 8:05 08 5 Mode of operation t: Duration of pulse, – Terminate the entering procedure. The parameters are effective immediately and do not have to be stored. Unit µs (microseconds). Can be set in increments of 5 µs between 0 and 15 µs–300 µs. n: Number of pulses 0–99. Can be set in one-pulse increments. Step 2 – Starting cell alignment – Trigger alignment U'~ by pressing the START key.
Multiporator_Text_en.fm Seite 77 Freitag, 9. Juni 2006 8:05 08 5 Mode of operation 5.3.2 Procedure for cell fusion Step 1 – Entering / modifying parameters – Press the SET 13:42 Example of a display of 12.AUG.00 electrofusion with activakey to select the U'~ 5V t 30s parameters desired U 30V t 15µs n2 ted alignments U'~ / U"~. (the parameter will U" ~ 5V t 30s be underlined). – Use the arrow 13:42 12.AUG.00 keys to modify the U'~ 0V t 30s parameter in the U 30V t 15µs n2 direction required.
Multiporator_Text_en.fm Seite 78 Freitag, 9. Juni 2006 8:05 08 5 Mode of operation Step 2 – Starting cell fusion – Trigger the cell fusion process by Pressing the START key. – "Charge" appears briefly in the fourth line of the display. Charging commences. 12.AUG.00 13:42 U'~ 5V t 30s U 30V t 15µs n2 Charge – During the charging procedure, 13:42 12.AUG.00 alternating voltage U'~ 5V t 30s U'~ is applied for the U 30V t 15µs n2 selected duration.
Multiporator_Text_en.fm Seite 79 Freitag, 9. Juni 2006 8:05 08 5 Mode of operation – After the discharging procedure has ended, the alternating voltage U"~ is applied for the set duration (0–95 seconds). The overall time remaining for the experiment is constantly visible on the bottom line of the display. After the experiment has 12.AUG.
Multiporator_Text_en.fm Seite 80 Freitag, 9. Juni 2006 8:05 08 6 Error messages In the event of an error, the Multiporator emits an acoustic signal and an error message appears in the display or via the printer (optional). These error messages refer to device errors or to applicational errors. Errors 1–4 and 9 may occur in the mode for bacteria, errors 1–7 in the mode for eukaryotic cells and errors 1–8 in the mode for cell fusion. Error Error message Cause no.
Multiporator_Text_en.fm Seite 81 Freitag, 9. Juni 2006 8:05 08 6 Error messages Error Error message Cause no. 07 Short τ The capacitor was completely emptied; it is uncertain whether the τ has been executed. The limiting element was the capacitor and not the pulse regulator. Solution Reduce the conductivity of the solution used. Check whether the correct cuvette type has been inserted. 08 Low Resistance The current in the cuvette is too high and the alternating voltage cannot be maintained.
Multiporator_Text_en.fm Seite 82 Freitag, 9. Juni 2006 8:05 08 7 Maintenance and servicing Warranties and servicing are the responsibility of the distributor. 7.1 Disinfection Before disinfecting the Multiporator, disconnect the device from the main power supply. All parts of the Multiporator, including the accessories and the connecting cable, must undergo wipe disinfection. The cuvette insert can also undergo spray disinfection.
Multiporator_Text_en.fm Seite 83 Freitag, 9. Juni 2006 8:05 08 8 Technical data Voltage/frequency: Fuses: Power consumption: Excess-voltage category: Protection class: Degree of contamination: Voltage range for the following modules: Eukaryotic cells: 100–240 V ±10 %, 50–60 Hz T0, 1.0 A – 5 x 20 mm (2 pcs.
Multiporator_Text_en.fm Seite 84 Freitag, 9. Juni 2006 8:05 08 8 Technical data Weight: Dimensions: max. 5.5 kg (according to modification level) Width: 25.2 cm Depth: 34.2 cm Height: 12.0 cm The device is -approved and has UL and CSA authorization (cUL: E 158089). U.S. Pat. No. 6,008,038.
Multiporator_Text_en.fm Seite 85 Freitag, 9. Juni 2006 8:05 08 9 Ordering information Order no. 4308 000.015 4308 000.023 4308 000.031 4308 000.040 Multiporator, basic version, for eukaryotic cells, 100–240 V, 50–60 Hz Multiporator, eukaryotic cells, and modification level "bacteria mode" Multiporator, eukaryotic cells and modification level "cell fusion" Multiporator, eukaryotic cells and modification level "bacteria mode" and "cell fusion" 4308 070.501 4308 070.510 4308 070.528 4308 070.
Multiporator_Text_en.fm Seite 86 Freitag, 9. Juni 2006 8:05 08 9 Ordering information 0013 608.164 6547 001.018 0013 610.517 0013 610.525 0013.610.533 4308 010.002 4308 011.
Multiporator_Text_en.fm Seite 87 Freitag, 9. Juni 2006 8:05 08 9a Ordering information for North America Order no.
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Multiporator_Text_sp.fm Seite 213 Freitag, 9. Juni 2006 8:13 08 EG-Konformitätserklärung EC Conformity Declaration Das bezeichnete Produkt entspricht den einschlägigen grundlegenden Anforderungen der aufgeführten EG-Richtlinien und Normen. Bei einer nicht mit uns abgestimmten Änderung des Produktes oder einer nicht bestimmungsgemäßen Anwendungverliert diese Erklärung ihre Gültigkeit. The product named below fulfills the relevant fundamental requirements of the EC directives and standards listed.
Titel_d_e_f_i_sp.fm Seite 3 Freitag, 9. Juni 2006 8:02 08 Eppendorf Offices ASEAN FRANCE SPAIN Eppendorf AG Regional Office in Malaysia Tel. +60 3 8023 2769 Fax +60 3 8023 3720 E-Mail: eppendorf@eppendorf.com.my Internet: www.eppendorf.com.my EPPENDORF FRANCE S.A.R.L. Tel. +33 1 30 15 67 40 Fax +33 1 30 15 67 45 E-Mail: eppendorf@eppendorf.fr Internet: www.eppendorf.fr Eppendorf Ibérica S.L. Tel. +34 91 651 76 94 Fax +34 91 651 81 44 E-Mail: iberica@eppendorf.de Internet: www.eppendorf.
Titel_d_e_f_i_sp.fm Seite 4 Freitag, 9. Juni 2006 8:02 08 Your local distributor: www.eppendorf.com/worldwide Eppendorf AG 22331 Hamburg · Germany Tel. +49 40 538 01-0 Fax +49 40 538 01-556 E-Mail: eppendorf@eppendorf.com Eppendorf North America, Inc. One Cantiague Road, P.O. Box 1019 Westbury, N.Y. 11590-0207 USA Tel. +1 516 334 7500 Toll free phone 800 645 3050 Fax +1 516 334 7506 E-Mail: info@eppendorf.com Application Support Europe, International: Tel. +49 1803 666 789 E-Mail: support@eppendorf.
