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User's Manual

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1. PES Figure 1 Operating Range for the 630 with Model 620 in LV with PBS at various volumes Operating Range for 640 PBS LV 4 mm gap cuvette 500 Re 1476 anm 20 to 500 1 uF 500 4 R 25 uh EU 4 25 to 70 msec E Kin 0 01 0 1 1 10 100 Pulse Ger KjI ims 01 A L 65D gn 8DO ul Figure 2 Operating Range for the 630 with Model 640 in LV with PBS at various volumes Page 24 www btxonline com ECM 630 Electroporation System Appendix A 630 Electrode Operation Ranges continued Operating Range 3B86 PBS LV gW 0 01 0 1 1 10 Pulsa Length jma 385 amp mL g 366 15 mL Figure 3 Operating Range for the 630 with 366 in LV with PBS at various volumes Operating Range for PP35 PBS LV Voltage v Pulse Length msi PP 35 2 g 1mL PBS 35 2 gp 3mL PBS Figure 4 Operating Range for the 630 with PP35 in LV with PBS at various volumes Operating Range for 6 amp 10 Water HV vVakamga v Pulse Length ms E3610 50 uL Figure 5 Operating Range for Model 610 H 0 HV Page 25 Petri dish electrode 20 to 350V 25 usec to 6 msec Petri Pulser 20 to 400 V 10 usec to 40 msec mm gap cuvette 50 to 2500 V 680 usec to 36 msec www btxonline com ECM 630 Electroporation Sy
2. 27 38 Appendix C Optimization Strategies 39 Appendix D Electrical Troubleshooting 40 Appendix E Experimental Troubleshooting 41 Appendix Glossary of Electrical Terms 42 Appendix G Glossary of Biological amp Technical Terms 43 44 Appendix H Electroporation Generator Compatibility 45 Appendix I Recommended Reading 46 Appendix J Accessories and Replacement Parts 47 Appendix General Care and Cleaning 48 Appendix L Connecting Electrodes 49 51 Appendix M Plate Handler 52 53 Page 1 www btxonline com ECM 630 Electroporation System General Information Serial Number The serial number for the 630 is located on the rear of the instrument case All inquiries concerning these products should refer to the serial numbers on the units Calibration There is no calibration required for the ECM 630 Warranty Harvard Apparatus warranties the 630 for a period of two years from the date of purchase At its option BTX Harvard Apparatus will repair or replace the unit if it is found to be defective as to workmanship or materials This warranty does not extend to any instrumentation which has been a subjected t
3. 4mm 25 Well Adapter HT 25 Well Disposable plate 2mm gap 25 Well Disposable Plates 2mm gap pkg 6 Enhancer 3000 Electroporation Monitoring System www btxonline com ECM 630 Electroporation System Appendix K General Care and Cleaning General Care Do not store or leave the instrument where the LCD display will be exposed to direct sunlight for long periods of time CAUTION To avoid damage to the instrument do not expose to sprays liquids or solvents Cleaning Inspect the instrument as often as operating conditions require To clean the instrument exterior perform the following steps 1 Remove loose dust on the outside of the instrument with a lint free cloth Use care to avoid scratching the clear plastic display filter 2 Usea soft cloth dampened with water to clean the instrument Use an aqueous solution of 7596 isopropyl alcohol for more efficient cleaning CAUTION To avoid damage to the surface of the instrument do not use any abrasive or chemical cleaning agents Use caution not to drop or cause any unwarranted physical harm to the instrument during any cleaning operations Page 48 www btxonline com ECM 630 Electroporation System Appendix Connecting Electrodes Cuvettes Safety Stand 1 Remove cuvette and the transfer pipette from their sterile packaging 2 Remove the cuvette cover and fill the cuvette with sample using the transfer pipette and replace cover
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6. 3134 375 3171 25 3208 125 3187 5 3225 32625 3240 525 3278 75 3316 875 3293 75 3332 5 3371 25 3346 875 3386 25 3425 625 www btxonline com 2200 55 110 185 220 275 330 385 440 495 550 680 715 825 880 935 990 1045 1100 1155 1210 1265 1320 1375 3490 1485 1540 1595 1650 1705 1760 1815 1870 1925 1980 2146 2200 2255 2310 2365 2420 2475 2530 2585 2640 2895 2750 2805 2850 2915 2970 3025 3080 3135 3190 3245 3300 3355 3410 2225 55 625 111 25 168 875 2225 278125 333 75 389 375 445 500625 556 25 811 875 8675 723 125 778 75 834 375 800 945 625 1001 25 1056875 11125 1168 125 122375 1279 375 1335 1300825 1446 25 1501875 15575 1613 125 1668 75 1724375 1780 1835 625 1891 25 1946 875 20025 2058 125 2113 75 2169375 2225 2280625 2336 25 2391875 24475 2503 125 2558 75 2814375 2670 2725 625 2781 25 2836 875 28925 2948 125 300375 3059 378 3115 3170625 3226 25 3281 875 33375 3393 125 344875 3504 375 ECM 630 Electroporation System 1600 880 920 980 1000 1040 1080 1120 1160 1200 1240 1280 1320 1350 1400 1440 1480 1520 1560 1600 1640 1680 1720 1760 1800 1840 1880 1920 1960 2040 2080 2120 2160 2200 2240 2280 2320 2360 2400 2440 2480 2520 uF 1625 1850 40625 4125 81 25
7. Transgene incorporation in which simple transfection of fish embryos has resulted in transgenic zebrafish Utilization of sperm as biological DNA carriers in which pulsed fields cause the complexing of DNA to sperm which then act as carriers upon fertilization Acrosome enhancement in which an exponential decay pulse enhances the acrosome reaction and facilitates fertilization Embryonic Stem Cell Chimeras in which embryonic stem cell transfection followed by micromanipulation into host blastomere has resulted in chimeric mice Parthogenesis in which a repetitive DC pulse stimulates an unfertilized egg to activate and divide as if fertilized resulting in haploid and diploid embryos www btxonline com ECM 630 Electroporation System Appendix A 630 Electrode Operation Ranges Figures 1 6 display expected operating ranges for the 630 with various chambers and electrodes The data lines in the following figures represent the arcing boundaries for the given electrode chamber model electroporation media volume and voltage mode The area at and above each line represents 100 arcing probability The area beneath each line represents parameters that may lead to the delivery of a full pulse This data was empirically determined in the BTX Application Laboratory Operating Range for 620 PBS LV 2mm gap cuvette 20 to 500V 25 usec to 93 msec voltage V Pulsa Langth ms 5620 40
8. User s Manual 630 Electroporation System 630 ro Cel Manipulator 9 45 0051 630 Electroporator only 110V 45 00511 ECM 630 Electroporator only 220V HARVARD APPARATUS The Electroporation Experts Publication 5501 003 REV E WEEE RoHS Compliance Statement EU Directives WEEE and RoHS To Our Valued Customers We are committed to being a good corporate citizen As part of that commitment we strive to maintain an environmentally conscious manufacturing operation The European Union EU has enacted two Directives the first on product recycling Waste Electrical and Electronic Equipment WEEE and the second limiting the use of certain substances Restriction on the use of Hazardous Substances RoHS Over time these Directives will be implemented in the national laws of each EU Member State Once the final national regulations have been put into place recycling will be offered for our products which are within the scope of the WEEE Directive Products falling under the scope of the WEEE Directive available for sale after August 13 2005 will be identified with a wheelie bin symbol Two Categories of products covered by the WEEE Directive are currently exempt from the RoHS Directive Category 8 medical devices with the exception of implanted or infected products and Category 9 monitoring and control instru ments Most of our products fall into either Category 8 or
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17. 2008 125 Resistor ohm Page 32 1300 32 5 87 5 130 162 5 195 227 5 260 292 5 325 357 5 390 422 5 455 487 5 520 552 5 585 617 5 650 682 5 715 747 5 780 812 5 845 877 5 910 942 5 975 1007 5 1040 1072 5 1105 1137 5 1179 1202 5 1235 1267 5 1300 1332 5 1365 1397 5 1430 1462 5 1527 5 1560 1592 5 1657 5 1690 1722 5 1755 1787 5 1820 1852 5 1917 5 1959 1982 5 2015 2047 5 1325 33 125 66 25 99 375 132 5 165 525 198 75 231 875 265 298 125 331 25 364 375 397 5 430 825 463 75 496 875 530 563 125 596 25 629 375 662 5 695 525 728 75 761 875 795 828 125 861 25 894 3 5 927 5 960 525 993 75 1026 875 1060 1093 125 1126 25 1159 375 1192 5 1225 625 1258 75 1291 875 1325 1358 125 1391 25 1424 375 1457 5 1490 525 1523 75 1556 875 1590 1623 125 1656 25 1689 375 1722 5 1755 525 1788 75 1821 875 1855 1888 125 1921 25 1954 375 1987 5 2020 525 2053 75 2086 875 1350 33 75 67 5 101 25 135 168 75 202 5 236 25 270 303 75 337 5 371 25 405 438 75 472 5 506 25 540 573 75 607 5 641 25 675 708 75 742 5 776 25 810 843 75 877 5 911 25 945 978 75 1012 5 1045 25 1080 1113 75 1147 5 1181 25 1215 1248 75 1282 5 1316 25 1350 1383 75 1417 5 1451 25 1485 1518 75 1552 5 1586 25 1620 1853 75 1687 5 1721 25 1755 1788 75 1822 5 1855 25 1890 1923 75 1957 5 1991 25 2025 2058 75 2092 5 2128 25 1375 34 375 68 75 103
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19. 375 25875 2830 625 2873 75 2716 875 1750 43 75 875 131 25 175 218 75 262 5 305 25 350 393 75 437 5 481 25 525 568 75 612 5 655 25 700 743 75 787 5 831 25 875 918 75 952 5 1008 25 1050 1093 75 1137 5 1181 25 1225 1268 75 1312 5 1356 25 1400 1443 75 1437 5 1531 25 1575 161875 1652 5 1705 25 1750 1793 75 1837 5 1881 25 1925 1968 75 2012 5 2055 25 2100 2143 75 2187 5 2231 25 2275 2318 75 2352 5 2405 25 2450 2493 75 2537 5 2581 25 2625 2668 75 2712 5 275825 1775 44 375 8B 75 133 125 1775 221 875 286 25 310 825 855 399 375 443 75 488 125 532 5 576 875 621 25 665 025 710 754 375 798 75 843 125 887 5 931 875 976 25 1020 825 1065 1109 375 115375 1198 125 1242 5 1288 875 1331 25 1375 625 1420 1464 375 1508 75 1553 125 1597 5 1641 875 1686 25 1730 625 1775 1819 375 1853 75 1908 125 1952 5 1998 875 2041 25 2085 825 2130 2174 375 221875 2263 125 2307 5 2351 875 2396 25 2440 625 2485 2529 375 257375 2518 125 2662 5 2706 875 2751 25 2795 825 1800 45 90 135 180 225 270 315 360 405 450 495 540 585 630 675 720 765 810 855 900 945 990 1035 1125 1170 1215 1260 1305 1350 1395 1440 1485 1530 1575 1620 1665 1710 1755 1800 1845 1890 1935 1980 2025 2070 2115 2160 2205 2250 2295 2340 2385 2430 2475 2520 2565
20. 375 1361 25 1423 125 1485 1546 875 1608 75 1670 825 1732 5 1794 375 1856 25 1918 125 1980 2041875 2103 75 2165 625 2227 5 2289 375 2351 25 2413125 2475 2536 875 2598 75 2660 625 2722 5 2784 375 2846 25 2908 125 2970 3031 875 3093 75 3155 625 3217 5 3279 375 3341 25 3403 125 3465 3526 875 3588 75 3650 625 3712 5 3774375 3836 25 3898 125 2500 62 5 125 187 5 250 312 5 375 437 5 500 5625 625 887 5 750 8125 875 9375 1000 1082 5 1125 1187 5 1250 1912 5 1375 1437 5 1500 1562 5 1625 1687 5 1750 1812 5 1875 1937 5 2000 2052 5 2125 21875 2250 2312 5 2375 2437 5 2500 25505 2625 2887 5 2750 28125 2875 2037 5 3000 30625 3125 3187 5 3250 3312 5 3375 3437 5 3500 3562 5 3625 3687 5 3750 3812 5 38 5 3937 5 2525 63 125 126 25 189 375 252 5 315 625 378 75 441 875 505 568 125 531 25 694 375 7575 820 825 883 75 946 875 1010 1073 125 1136 25 1199 375 1262 5 1325 825 1388 75 1451 875 1515 1578 125 1641 25 1704 375 1767 5 1830 625 1893 75 1956 875 2020 2083 125 2146 25 2209 375 2272 5 2335 825 239875 2461 75 2525 2588 125 2851 25 2714 375 2777 5 2840 825 2903 75 2986 575 3030 3093 125 3156 25 3219 375 3282 5 3345 525 3408 75 3471 875 3535 3598 125 3661 25 3724 375 3787
21. 5 584 375 59125 618 125 671 875 698 75 1316 875 1343 75 1370 825 1397 5 1424 375 145125 1478 125 1505 1831 875 1558 75 1585 625 1612 5 1639 375 186625 1100 275 55 825 110 137 5 165 192 5 247 5 302 5 357 5 4125 5775 715 7425 11275 1155 11825 1237 5 1265 12025 1320 13475 1375 1402 5 14575 15125 1567 5 1565 1822 5 15775 17325 1125 28125 56 25 84 375 1125 168 75 195 875 81875 646 875 703 125 73128 759 375 787 5 815625 871 875 956 25 984 375 1012 5 1088 75 1095 875 1125 1153125 1181 25 1208 375 12375 1293 75 1321 875 1406 25 1434 375 1462 5 1518 75 1545 875 1803 125 1631 25 1859 375 1687 5 1715 625 174375 1771875 517 5 575 603 75 632 5 651 25 718 75 7475 1083 78 1092 5 1121 25 1150 117875 175875 1782 5 1811 25 www btxonline com 1175 29 375 58 75 88 125 117 5 17625 205 625 293 75 323 125 3525 381 875 411 25 851 875 881 25 910 625 940 969 375 398 75 1028 125 1057 8 1086 875 1116 25 1145 625 1175 1204 375 1233 75 1263 125 1282 5 1321 875 1351 25 1380 625 1439 375 1468 75 1408 125 1527 5 1556 875 1586 25 1615625 1645 1674 375 1703 75 1733 125 1762 5 1701 875 1821 25 1850 625 8 8888355855588388 33 1410 1440 1470 1500 1530 1560 1590 1620
22. 525 270 292 5 315 337 5 360 382 5 405 427 5 450 472 5 495 517 5 540 2775 3006258 32375 346 875 370 393125 41625 439375 4525 485625 50875 531875 555 285 30875 332 5 355 25 380 403 75 4275 45125 475 49875 5225 54625 570 2925 316 875 341 25 365825 390 414375 438 75 483 125 4875 511 875 535 25 580 825 585 300 325 350 375 400 425 450 475 500 525 550 575 600 3075 333 125 35875 384 375 410 435625 451 25 486 875 5125 538125 56375 589 375 615 315 341 25 367 5 393 75 420 446 25 4725 49875 525 55125 5775 60375 630 3225 349 375 37825 1403 125 430 456875 48379 510825 5375 5643758 591 25 518125 845 330 357 5 385 4125 440 467 5 495 522 5 550 577 5 605 632 5 660 347 5 365 525 39375 421875 450 478 125 506 25 534 375 5625 590 625 618 75 545 875 675 345 37375 4025 431 25 460 488 75 5175 54625 575 60375 6325 651 25 680 3525 381875 41125 440625 470 499375 528759 558125 5875 818875 645259 875625 705 360 390 420 450 480 510 540 570 600 830 660 690 720 3675 3984125 42875 459 375 490 520625 551 06 581 875 6125 643125 67375 704 375 735 375 408 25 437 5 468 75 500 531 25 5625 59375 625 65625 6875 71875 750 3825 4143758 44825 478125 510 5418 5 57375 805525 6375 869375 70125 733125 765 390 4225 455 487 5 520 552 5 585 617 5 650 682 5 715 747 5 780 397 5 430 525 46375 496 875 5380 563125 59625 629 375 6525 6956258 72875 761 875 795 405 438 75 472 5 506 25 540 573 75 6075 641 25 675 70875 742 5 776 25 810 4125 4468 5 48125 515625 550 584375 81
23. 82 5 121875 12375 182 5 165 203 125 20625 24375 2415 284375 28875 325 330 365 525 37125 408 25 412 5 446 875 45375 487 5 495 528 125 53625 568 75 577 5 609 375 61875 650 660 690625 70125 731 25 742 5 771 875 78375 8125 825 853 125 88625 89375 907 5 934375 94875 975 990 1015625 1031 25 105625 10725 1096 875 1113 75 1137 5 1155 1178125 1196 25 121875 1237 5 1259 375 127875 1300 1820 1340 625 1351 25 138125 14025 1421875 144375 1452 5 1485 1503 125 152625 1543 75 15675 1584 375 160875 1625 1650 1685625 189125 170825 47325 1746 875 177375 1787 5 1815 1828 125 185625 188875 18975 1909 375 193875 1950 1980 1990625 2021 25 203125 2062 5 2071875 210375 2112 5 2145 2153 125 218625 219375 2227 5 2234375 225875 2275 2310 2315625 235125 235825 23925 2386815 243375 2437 5 2475 2478 125 251625 251875 2557 5 2559 375 2598275 Page 35 1875 41 875 8375 125 625 187 5 209 375 25125 293 125 335 375 875 418 75 460625 502 5 544 375 585 25 628 125 670 711 875 753 75 795625 837 5 879 375 92125 963125 1005 1048 875 1088 75 1130625 11725 1214375 1256 25 1298 125 1340 1381875 1423 75 1465 625 1507 5 1549375 1581 25 1633 125 1675 1718875 1758 75 1800 625 1842 5 1884 375 1928 25 1968 125 2010 2051 875 2093 75 2135 625 2177 5 2219375 2251 25 2303 125 2345 2385 875 2428 75 2470625 25125 2554 375 2596 25
24. If using the Enhancer 3000 to monitor output connect the banana plugs into the output ports of the High Voltage Probe Use the black and red high voltage cables to connect the BTX generator to the input ports of the Enhancer 3000 3 Open the Safety Stand cover 4 Use the thumb wheel to slide the electrodes open Page 49 www btxonline com ECM 630 Electroporation System Appendix Connecting Electrodes Continued 5 Place the Flat Pack Chamber in between electrodes with the aluminum coming in contact with the electrodes 6 Secure the Flat Pack Chamber in place by closing the gap with the thumb wheel The Flat Pack Chamber should be snugly placed between the electrodes however it should be loose enough that it can be pulled out without adjusting the thumb wheel again 7 Following instructions for the BTX generator set the appropriate parameters 8 Deliver the electroporation pulse s to the sample Warning Use proper eye protection during electroporation 9 Remove Flat Pack Chamber and extract the cell solution 10 Dispose of Flat Pack Chamber and prepare for the next experiment Flat Electrodes 1 Fill the sterile Flat Electrode Chamber using a syringe 2 Replace the clear plastic tip so the holes in the top align with the holes in the electrode bars 3 Push the banana plugs at the opposite end of the Flat Electrode Catalog number45 0217 into the voltage output of the BTX generator Warning
25. In this case call BTX Technical Support EEPROM Failure The unit has detected a malfunction in its internal memory system The validity of the data might be compromised Turning or pressing the knob will bring the Set Parameters screen Verify carefully every setpoint before pulsing This verification is performed during power up and every time that data is loaded from memory Contact BTX Technical Support if this error message is displayed again after a power up sequence Pulsing Aborted Charge Failure The unit did not charge its selected capacitor bank Turn or press the encoder knob Disconnect the load from the HV connector Select a different capacitor and press the pulse button Contact BTX Technical Support if a similar message is displayed again Pulsing Aborted Charging Timed Out A charging time limit of 20 seconds is provided for circuit safety If the capacitors are not charged to the pre set voltage level after 20 seconds the PULSING ABORTED CHARGING TIMED OUT message is displayed For assistance with this situation please contact BTX Technical Support Press the encoder or the pulse switch once to get back to the Set Parameters screen Page 40 www btxonline com ECM 630 Electroporation System Appendix E Experimental Troubleshooting Arcing Verify electrical component functionality Verify properties of cell sample do cells need to be washed Is the buffer appropriate for application Verify properti
26. Make sure the Generator is turned off prior to connecting any cables to it If using the Enhancer 3000 to monitor output connect the banana plugs into the output ports of the High Voltage Probe Use the black and red high voltage cables to connect the BTX generator to the input ports of the Enhancer 3000 4 Following instructions for the BTX generator set the appropriate parameters 5 Deliver the electroporation pulse s to the sample Warning Use proper eye protection during electroporation 6 Extract the cell solution The chamber can be rested on ice for cooling purposes 7 Clean as appropriate and prepare for the next experiment Page 50 www btxonline com ECM 630 Electroporation System Appendix Connecting Electrodes Continued Petri Dish Electrode 1 Plug the HV cables from the Petri Dish Electrode into the voltage output of the BTX Generator Warning Make sure the Generator is turned off prior to connecting any cables to it If using the 3000 to monitor output connect the banana plugs into the output ports of the High Voltage Probe Use the black red high voltage cables to connect the BTX generator to the input ports of the Enhancer 3000 Prepare sample for electroporation 35mm Petri Dish Place the electrode in the 35mm Petri Dish Allow it to gently rest on the surface of the dish 4 Following instructions for the BTX generator set the appropriate parameters Deli
27. The 630 is capable of reproducing most exponential competitive system protocols BioRad Gene Pulser and Gene Pulser II Protocols For protocols using the Capacitance Extender Capacitance Extender or the Capacitance Extender Plus use the resistance setting none reduce the voltage by 50 V and use the capacitance setting outlined in the protocol For protocols using the Pulse Controller Pulse Controller Il or Pulse Controller Plus use a capacitance setting of 25 pF reduce the voltage 50 V and use the resistance settings outlined in the protocol Optimization of voltage as outlined in Appendix C is recommended For additional recommendations please contact BTX Technical Support Other Competitive Exponential Decay Electroporator Protocols Attempt to identify the resistance and capacitance as well as the volt age outlined in the protocol Match these as closely to settings allowed with the 630 For low voltage protocols V 500V if there is no resistance outlined use the ECM 630 resistance setting For additional recommendations please contact BTX Technical Support Page 19 www btxonline com ECM 630 Electroporation System Advanced Operation Programming 1 2 Page 20 Set parameters as outlined in Operating Basics Once presetting of parameters has been completed rotate the parameter control knob until the save function has been reached Press knob in a
28. both the parameter under control and the value of the parameter under control 1 The display will indicate which parameter is under control by the presence of an arrow to the left of the parameter 2 select a parameter to adjust rotate the knob until the arrow is to the left of the desired parameter then push to select The arrow will move to the right of the value displayed for that parameter 3 To adjust the value of a parameter under control rotate the knob clockwise to increase the value and counter clockwise to decrease it Once values are adjusted push the knob to lock settings and arrow will return to left of parameters under control 4 In order to move between screens move the cursor to the bottom of the screen and rotate the knob clockwise to move to the next screen 5 In order to move to the previous screen move the cursor to the top of the screen and rotate the knob counterclockwise Display The ECM 630 Display will show two possible screens the Set Parameter Screen and the Pulse Delivered screen as well as various Status Messages Set Parameter Screen 1 The Set Parameters screen consists of two pages showing Set Parameters on the first line of each page 2 The Voltage Resistor Resistance and Capacitor Capacitance are shown on the first page and the Advanced Features including Save View and Load are shown on the second page of the Set Parameters screen 3 The 630 will
29. dyes sugars and dNTP S Electroporation has been used to study cellular activation processes by electropermeabilizing cells to Ca Mg and Electroporation is also used to electroinsert proteins into the cell membrane Finally electroporation has been used to introduce drugs such as the chemotherapeutic agent bleomycin into cancer cells in vitro and in vivo The use of low impedance buffers such as PBS may result in a voltage drop so that the actual peak voltage delivered to samples may be less than the set voltage With exponential decay generators monitoring is necessary to identify the pulse length or time constant since this parameter may be very much dependent on the impedance of the sample sample load When using complex and custom electroporation applicators and chambers the electroporation waveform may be altered and monitoring is again strongly recommended Molecules Introduced by Electroporation DNA siRNA miRNA 5 Enzymes Antibodies e Other Proteins e Peptides e Dyes Sugars e lons e Other Molecules High GFP expression in Mouse PE501 www btxonline com ECM 630 Electroporation System Applications continued Plant Protoplast Electroporation Electroporation has been used to introduce molecules into plant protoplasts pollen and most recently direct transfer into plant tissue in vivo Other Electroporation Applications 1 Page 23
30. per well b Press pulse on the electroporator to initiate the pulse s for that column the plate handler will beep c The column LEDs will flash to indicate the active column Once 3 column has been electroporated the LED remains blinking until the cover is opened NOTE Once the cover is opened the memory of pulsed columns if wiped out d Press the column select button to switch to the next column Plate handlers may be used with both types of HT plates HT 96 and HT 25 The following will outline the steps needed to configure the unit for the type of plate being used 630 lli Page 53 www btxonline com ECM 630 Electroporation System HARVARD APPARATUS The Electroporation Experts 84 October Hill Road Holliston MA 01746 Phone 508 893 8999 Toll Free 800 272 2775 U S Only Fax 508 429 5732 E mail techsupport btx harvardapparatus com Web www btxonline com Page 54 www btxonline com
31. read 5 The first page of the Set Parameter Screen from the last time the 630 was used will be displayed each time the 630 is powered up after this initial start up Page 10 www btxonline com ECM 630 Electroporation System Quick Start Continued Instrument Controls Pulse Switch Display High Voltage Parameter Power Output Control Knob Switch Power Switch 1 Electronic Power Switch located on the lower right front panel 2 Press once to initialize the ECM 630 and once more to turn off CAUTION A stabilization period of 3 seconds is required after connecting to an outlet Pulse Switch 1 The electronic Pulse Switch on the upper left front panel is activated in the ready mode when the Set Parameters screen is displayed 2 Once the start switch is activated the generator will charge the capacitors to the preset voltage then automatically deliver the pulse 3 The maximum charge time is line voltage dependent and will typically be less than 10 seconds 4 A pulse sequence may be aborted by pressing the Pulse switch a second time before the charging is completed 5 Following the delivery of a pulse the Pulse switch can be pressed once to leave the feedback mode and return to the ready mode Page 11 www btxonline com ECM 630 Electroporation System Quick Start Continued Parameter Control Knob The Parameter Control Knob is rotary encoder controlling
32. 0 160 170 180 190 200 210 220 230 240 1275 138 125 14875 159 375 170 180625 19125 201 875 2125 223125 23375 244 375 255 135 148 25 157 5 168 75 180 191 25 2025 21375 225 23625 2475 25875 270 1425 154 375 168 205 178 125 190 201 875 213 75 225 625 2375 249 375 261 25 273 125 285 150 162 5 175 187 5 200 2125 225 237 5 250 262 5 275 287 5 300 1575 170625 18375 196875 20 223125 23625 249375 2525 275625 28875 301 875 315 165 178 75 1825 205 25 220 233 75 2475 251 25 275 28875 3025 31625 330 1725 186 875 201 25 215 625 230 244 375 258 75 273 125 2875 301 875 316 25 330 925 345 180 195 210 225 240 255 270 285 300 315 330 345 360 1875 203 125 21875 234 375 250 285 625 281 25 296875 3125 328125 34375 359 375 375 195 211 25 227 5 243 75 260 276 25 29258 30875 325 341 25 3575 37375 390 2025 219375 23625 253 125 270 286 275 30375 320 625 3375 3543759 371 25 388125 405 210 227 5 245 2625 280 2975 315 3325 850 367 5 385 402 5 420 217 5 235625 25875 271 875 280 308125 32625 344375 3525 3806259 39875 416 875 435 225 24375 2825 281 25 300 318 75 3375 35625 375 39375 4125 43125 450 2325 2518 5 27125 290625 310 329375 348 75 388125 3875 406875 428 258 445 825 465 240 260 280 300 320 340 360 380 400 420 440 460 480 2475 268 125 28875 309 375 330 350625 37125 391 875 4125 433125 45375 474375 495 255 278 25 297 5 318 75 340 381 25 3825 40375 425 44625 46875 488 75 510 2825 284375 30625 328 125 350 371 875 393 75 415 625 4375 459 375 481 25 503 125
33. 1 875 17875 185 625 192 5 199 375 206 25 213 125 220 226 875 233 75 240 625 247 5 254 375 261 25 268 125 275 281 875 288 75 295 625 302 5 308 375 316 25 323 125 330 336 875 343 75 350 625 357 5 364 375 371 25 378 125 385 391 875 39875 405 625 4125 419 375 42625 433 125 ECM 630 Electroporation System 300 325 350 375 400 425 450 475 500 525 550 575 500 7 5 8 125 8 75 9375 10 10 625 1125 11 875 125 13 125 13 75 14 375 15 15 16 26 175 18 75 20 21 26 225 23 75 25 26 25 27 5 28 75 30 225 24375 26 25 28 125 30 31 875 3375 35625 375 39 375 41 25 43425 45 30 32 5 35 37 5 40 425 45 475 50 52 5 55 57 5 60 375 40625 43 75 46 875 50 53125 56 25 59375 825 65625 68 75 71 875 75 45 48 75 52 5 56 26 60 63 75 875 71 25 75 78 75 825 86 25 90 52 5 56 875 61 25 55 625 70 74 375 78 75 83 125 875 91 875 96 25 100625 105 60 65 70 75 80 85 90 95 100 105 110 115 120 67 5 73 125 78 75 84 375 90 95625 10125 106875 1125 118125 123 258 129 375 135 75 81 25 87 5 93 75 100 106 25 112 5 118 75 125 131 25 137 5 143 75 150 82 5 89 375 9625 102 125 110 116875 123 75 130 625 137 5 144 375 151 258 158 125 165 90 875 105 125 120 1275 135 1425 150 157 5 165 172 5 180 975 105625 1375 121 875 130 138125 14625 154375 1825 170625 17875 185 875 195 105 113 75 122 5 131 25 140 148 75 157 5 166 25 175 183 75 192 5 201 25 210 1125 121 875 131 25 140 625 150 159 375 168 75 178 125 187 5 196 875 206 25 215 625 225 120 130 140 15
34. 118 75 1425 190 21375 6175 102125 1088 75 1116 25 1140 1183 75 1187 5 1211 25 1235 1258 75 1282 5 1306 25 1336 1353 75 13775 1401 25 1448 76 14725 1496 25 975 24 375 48 75 73 125 97 5 121875 146 25 179 625 29375 243 75 268 125 292 5 315 875 341 25 414375 463 125 4875 511 875 609 375 633 75 658 125 682 5 731 25 804 375 82875 853 25 877 5 901 875 926 25 950 625 999 375 1023 75 1048 125 1072 5 1121 25 1145 625 1170 1194 375 1218 75 1243 125 1267 5 1291875 1316 25 1340 625 1389 375 1413 75 1438 125 1462 5 1486 875 1511 25 1535 625 x SERSSSELSSSRSASTSSSSRSSARERARSESBRESERSSZReaees A m 5883288838383 1025 5125 461 25 486 875 5125 538 125 563 75 588 375 615 640 625 821 875 717 5 87125 856 875 97375 1101 875 1127 5 1153 125 1178 75 1204 375 1230 1255 625 128125 1306 875 1332 5 1358 125 1383 75 1408 375 1435 1450 625 1485 25 1511875 1537 5 1583 125 1588 75 1814 375 1050 28 25 52 5 78 75 105 131 25 1575 183 75 288 75 315 341 25 357 5 393 75 445 25 472 5 498 75 55125 813 75 866 25 918 75 1338 75 1365 1391 25 1417 5 1443 75 1470 1495 25 1522 5 1548 75 1575 1501 25 1627 5 1853 75 1075 26 875 53 75 80 525 107 5 134 375 161 25 188 125 215 241 875 268 75 3225 349 375 403 125 483 75 537
35. 125 1375 171 875 20825 240 525 275 309 375 34375 378 125 4125 446 875 481 25 515 825 550 584 375 618 75 853 125 6875 721875 758 25 790 525 825 859 375 80375 928 125 962 5 996 875 1031 25 1065 525 1100 1134 375 1168 75 1203 125 1237 5 1271 875 1306 25 1340 525 1375 1409 375 144375 1478 125 1512 5 1546 875 1581 25 1815 825 1650 1684 375 1718 75 1753 125 1787 5 1821 875 1856 25 1890 825 1925 1959 375 1993 75 2028 125 2062 5 2096 875 2131 25 2185 825 595 630 700 735 770 805 840 875 910 945 980 1015 1050 1085 1120 1155 1190 1225 1260 1295 1330 1365 1400 1435 1470 1505 1540 1575 1610 1645 1680 1715 1750 1785 1820 1865 1890 1925 1960 1995 2030 2100 2135 2170 2205 1425 35 625 71 25 106 875 1425 178125 21375 249 375 285 320 625 356 25 391 875 4275 463 125 49875 534375 570 805 625 641 25 676 875 71 25 748 125 783 75 819 375 855 850 625 926 25 961 875 9875 1033 125 1068 75 1104 375 1140 1175625 121 125 1248 875 12825 1318 125 1353 75 1389 375 1425 1460 625 1498 25 1531 875 1567 5 1803 125 1638 75 1874 375 1710 1745 825 1781 25 1816 875 1852 5 1888 125 1923 75 1959 375 1995 2030 625 2086 25 2101 875 2197 5 2173 12
36. 1650 1680 1710 1740 1770 1800 1830 1860 1890 1225 30 525 61 25 61 575 122 5 153 125 183 75 214 375 275 825 308 25 336 875 367 5 398 125 428 75 459 375 520 525 551 25 581 875 843 125 873 75 704 375 765 625 796 25 826 875 857 5 888 125 918 75 1010 525 1041 25 1071 875 1102 5 1134 125 1163 75 1194375 1225 1255 525 1286 25 1316 875 13475 1378 125 1408 75 1439 375 1500 525 1531 25 1561 875 1592 5 1823 125 1653 75 1684 375 1715 1745 525 1775 25 1806 875 1837 5 1868 125 1898 75 1929 375 312 937 a 156 25 2187 2812 312 3437 468 75 5312 593 7 656 2 7187 781 8437 937 968 75 1081 2 1062 1093 7 1156 2 1187 1218 75 1281 2 1343 75 1406 2 1437 1468 15312 1593 7 1656 2 17187 1 n 1781 2 1812 5 1843 7 1906 25 1968 7 ECM 630 Electroporation System Appendix 630 RC Time Constants Continued Low Voltage Mode RC 1275 31 875 63 75 95 625 127 5 159 375 191 25 223 125 255 286 875 318 75 350 525 382 5 414 375 446 25 478 125 510 541 375 573 75 605 525 637 5 669 375 701 25 733 125 765 796 375 828 75 860 525 892 5 924 375 956 25 988 125 1020 1051 875 1083 75 1115 825 1147 5 1179 375 1211 25 1243 125 1275 1306 875 1338 75 1370 625 1402 5 1434 375 1466 25 1498 125 1530 1561 875 1593 75 1625 825 1657 5 1689 375 1721 25 1753 125 1785 1816 875 1848 75 1880 625 1912 5 1944 375 1976 25
37. 1687 5 1734 375 1781 25 1828 125 1875 1921 875 196875 2015 625 2082 5 2109 3 5 2156 25 2203 125 2250 2296 875 2343 75 2390 625 2437 5 2484 375 2531 25 2578 125 2625 267 1 875 271875 2765 525 28125 2859 375 2906 25 2953 125 www btxonline com 1900 47 5 95 142 5 190 237 5 285 332 5 380 427 5 475 522 5 570 617 5 665 712 5 760 807 5 855 902 5 950 997 5 1045 1092 5 1140 1187 5 1235 1282 5 1330 1377 5 1425 1472 5 1520 1567 5 1615 1662 5 1710 1757 5 1805 1852 5 1900 1947 5 1995 2042 5 2090 2137 5 2185 2232 5 2280 2327 5 2375 2422 5 2470 2517 5 2565 2612 5 2660 2707 5 2753 2802 5 2850 2897 5 2945 2992 5 ECM 630 Electroporation System Appendix ECM 630 RC Time Constants Continued Low Voltage Mode RC 1925 48 125 96 25 144 375 1925 240625 288 75 336 875 385 433425 481 25 529 375 5775 625 625 673 75 721 875 818 125 886 25 914375 9625 1010 625 1058 75 1106 875 1158 1203 125 1251 25 1299 375 13475 1395 625 144375 1491 875 1540 1588 125 163825 1684 375 1732 5 1780 625 1828 75 1876 875 1925 1973 125 2021 25 2069 375 2117 5 2165 625 2213 75 2261 875 2310 2358 125 2405 25 2454 375 2502 5 2550 625 2598 75 2646 875 2695 2743 125 2791 25 2839 375 2887 5 2935 625 208375 3031 875 Resistor ohm Page 34 1950 48 75 975 146 25 195 243 75 2925 341 25 390 43875 487 5
38. 2 5 2055 25 2100 2143 75 2187 5 2231 25 2275 2318 75 2352 5 2405 25 2450 249375 2537 5 2581 25 2625 2668 75 2712 5 2758 25 1775 44 375 8875 133 125 177 5 221 875 266 25 310 625 355 399 375 443 75 488 125 532 5 576 875 521 25 665 625 154 375 798 75 843 125 887 5 931 875 976 25 1020 825 1065 1109 375 1153 75 1198 125 1242 5 1285 875 1331 25 1375 825 1420 1464 375 1508 75 1553 125 1597 5 1641 875 1686 25 1730 625 1775 1819 375 1883 75 1908 125 1952 5 1996 875 2041 25 2085 625 2130 2174 375 2218 75 2263 125 2307 5 2351 875 2396 25 2440 625 2485 2529 375 2573 75 2618 125 2662 5 2705 875 2751 25 2785 825 1800 45 90 135 180 225 270 315 360 405 450 495 540 585 530 875 720 765 1825 45 625 91 25 136 875 182 5 228 125 273 75 319 375 365 410 625 456 25 501 875 547 5 593 125 538 75 884 375 730 775 625 821 25 856 875 9125 958 125 1003 75 1049 375 1095 1140 625 1186 25 1231 875 1277 5 1323 125 1368 75 1414 375 1460 1505 625 1551 25 1596 875 16425 1688 125 1733 75 1779 375 1825 1870 625 1916 25 1951875 2007 5 2053 125 2098 75 2144 375 2190 2035625 2281 25 2326 875 2372 5 2418125 2463 75 2509 375 2555 2620 625 2691 875 2737 5 2783 125 2828 75 28 4 375 1850 4625 925 138 78 185 231 25 277 5 323 75 370 415 25
39. 2137 5 2208 75 2280 2351 25 2422 5 2493 75 2565 2636 25 2707 5 2778 75 2850 2921 25 2992 5 3063 75 9135 3206 25 3277 5 3348 75 3420 3401 25 3582 5 3833 75 3705 3776 25 3847 5 3018 75 3990 4061 25 4192 5 4203 75 4275 4346 25 4417 5 4488 75 2875 71 875 143 75 215 625 287 5 359 375 431 25 503 125 575 646 875 71875 790 625 852 5 934 375 1008 25 1078 125 1150 1221 875 123 75 1385 825 1437 5 1509 375 1581 25 1653 125 1725 1796 875 1868 75 1940 625 2012 5 2084 375 2156 25 2228 125 2300 2371 875 2443 75 2515 625 2587 5 2659 375 2731 25 2803 125 2875 2946 875 3018 75 3090 625 3152 5 3234 375 3306 25 3378 125 3450 3521 875 3593 75 3655 625 3737 5 3809 375 3881 25 3953 125 4025 4096 875 4168 75 4240 825 4312 5 4384 375 4456 25 4528 125 ECM 630 Electroporation System 2250 56 05 112 5 168 75 225 281 25 337 5 393 75 450 506 25 5625 618 75 675 731 25 787 5 84375 900 956 25 10125 1068 75 1125 118125 1237 5 1293 75 1350 1406 25 1462 5 1518 75 1575 1631 25 1687 5 1743 75 1800 1856 25 1912 5 1968 75 2025 2081 25 21315 219375 2250 2306 25 2362 5 2418175 2475 2531 25 2587 5 2643 75 2700 2756 25 2812 5 2868 75 2925 2981 25 30375 3093 75 3150 3206 25 30625 3318 75 3375 343125 3487 5 3543 75 2275 56 875 113 75 170 625
40. 25 257 5 321 875 366 25 450 525 515 579 375 643 75 708 125 772 5 836 575 901 25 965 525 1030 1094 375 1158 75 1223 125 1287 5 1351 875 1416 25 Resistor 1480 525 1545 1609 375 1673 75 1738 125 1802 5 1866 875 1931 25 1995 525 2060 2124 375 218875 2253 125 2317 5 2381 875 2446 25 2510 625 2575 2639 375 2703 75 2768 125 _ 28325 2896 875 2961 25 3025 625 3090 3154 375 3218 75 3283 125 3347 5 3411 875 3476 25 3540 625 3605 3669 375 3733 75 3798 125 3862 5 3926 875 3991 25 4055 825 Page 36 2600 85 130 195 260 325 390 455 520 585 650 715 780 845 910 975 1940 1105 1170 1235 1300 1385 1430 1560 1625 1690 1755 1820 1885 1959 2015 2080 2145 2210 2275 2340 2405 2470 2535 2609 2665 2730 2795 2860 2925 2990 3055 3120 3185 3250 3315 3380 3445 3510 3575 3640 3705 3770 3835 3909 3965 4030 4095 2625 85 625 131 25 196 875 2625 328 125 393 75 459 375 525 590 525 656 25 721875 787 5 853 125 91875 984 375 1050 1115 525 1181 25 1246 875 13125 1378 125 144375 1509375 1575 1640 825 1708 25 1771 875 1837 5 1903 125 196875 2034 375 2100 2165 625 2231 25 2296 875 23625 2428 125 2493 75 2559 375 2625 2690 525 2758 25 2821 875 _ 28875 2953 125 301875 3084 375 3150 3215 825 3281 25 3346 875 34125 3478 125 3543 75 3609 375 3875 3740 625 3808 25 3871 875 3937 5 4003 125 406
41. 2610 2655 2700 2745 2 90 2835 1825 45 825 91 25 136 875 1825 228125 273 75 319 375 365 410 525 456 25 501 875 547 5 593 125 538 75 584 375 730 775 625 821 25 866 875 9125 958 125 1003 75 1049 375 1095 1140 625 1186 25 1231 875 1277 5 1323 125 1368 75 1414 375 1460 1525 625 1551 25 1596 875 1642 5 1688 125 1733 75 1779 375 1825 18 0 625 1916 25 1981 875 2007 5 2053 125 2098 75 2144375 2190 2235 605 2281 25 2326 875 2372 5 2418 125 2463 75 2509 375 2555 2600 625 2646 25 2691 875 2737 5 2783 125 2828 75 2874 375 1850 46 25 925 138 75 185 231 25 277 5 323 75 370 41625 4625 508 75 555 501 25 647 5 593 75 740 786 25 8325 878 75 925 971 25 1017 5 1063 75 1110 1156 25 12025 1248 75 1295 1341 25 1387 5 143375 1480 1526 25 1572 5 1618 75 1665 171125 1757 5 1803 75 1850 1898 25 1942 5 1988 75 2035 2081 25 2127 5 217375 2220 2266 25 2312 5 2358 75 2405 2451 25 2497 5 2543 75 2590 2535 25 2682 5 279875 2775 2821 25 2857 5 2913 75 1875 46 875 9375 140825 1875 234 375 281 25 328 125 375 421 875 468 75 515 625 562 5 609 375 656 25 703 125 750 796 875 843 75 890 625 9375 984 375 1031 25 1078 125 1125 1171 875 1218 75 1265 825 1312 5 1359 375 1406 25 1458 125 1500 1546 875 1593 75 1640 525
42. 3 Place cuvette on ice for cooling purposes Push the banana plugs of the Safety Stand into the voltage output of the BTX generator Warning Make sure the Generator is turned off prior to connecting cables to it If using the Enhancer 3000 to monitor output connect the banana plugs into the output ports of the High Voltage Probe Use the black and red high voltage cables to connect the BTX generator to the input ports of the Enhancer 3000 5 Open the Safety Stand cover Use the thumb wheel to slide the electrodes open Place a BTX cuvette in between the electrodes with the aluminum of the cuvette coming in contact with the electrodes 8 Secure the cuvette in place by closing the gap with the thumb wheel The cuvette should fit snugly between the electrodes however it should be loose enough that it can be pulled out without adjusting the thumb wheel again 9 Following instructions for the BTX generator Set the appropriate parameters 10 Deliver the electroporation pulse s to the sample Warning Use proper eye protection during electroporation 11 Remove cuvette cover and extract the cell solution 12 Dispose of cuvette and prepare for the next experiment Flat Pack Chambers 1 Fill the flat pack chamber with sample using a pipette 2 Push the banana plugs of the Safety Stand into the voltage output of the BTX generator Warning Make sure the Generator is turned off prior to connecting any cables to it
43. 31 25 e 650 00 16 25 82 50 875 00 16 88 33 75 259526 SDN m 700 00 17 50 35 00 2175875 219375 E TERI 1518 CES 2257 5 2275 L 750 00 18 75 37 50 2338 125 2356 25 775 00 19 38 38 75 241815 2437 5 5 800 00 20 00 40 00 2499375 251875 to 825 00 20 63 41 25 2580 2600 850 00 21 25 42 50 2650 625 2681 25 875 00 21 88 43 75 2741 25 2762 5 d 900 00 22 50 45 00 si 925 00 23 13 46 25 S 950 00 23 75 47 50 ems 975 00 24 38 48 75 3144 375 3168 75 1909 00 ab 1025 00 25 63 51 25 3305825 333125 1050 00 26 25 52 50 3386 25 34125 3 1075 00 26 88 53 75 3468 875 3493 75 1100 00 27 50 55 00 3547 5 3575 1125 00 28 13 56 25 3628 125 3656 25 1150 00 28 75 57 50 3708 75 3737 5 1175 00 29 38 58 75 3789 375 3818 75 1200 00 30 00 60 00 3870 3900 1225 00 30 63 61 25 3900 605 1250 00 31 25 62 50 e 1275 00 31 88 63 75 s pec 1300 00 82 50 65 00 e 1325 00 33 13 66 25 pue 1350 00 33 75 67 50 adus 1375 00 34 38 68 75 4515 4550 1400 00 35 00 70 00 4595 625 4631 25 1425 00 35 63 71 25 4676 25 47125 1450 00 36 25 72 50 4758875 479375 i 1475 00 36 88 73 75 4837 5 4875 1500 00 37 50 75 00 4918 125 4956 25 1525 00 38 13 76 25 4998 75 5037 5 1550 00 88 75 77 50 S 11 1575 00 39 38 78 75 Page 38 www btxonline com ECM 630 Electroporation System Appendix C Optimization Strategies General The success of electro cell ma
44. 375 1118625 1147 5 8125 845 8775 910 942 5 975 1007 5 1040 1072 5 1105 1137 5 1170 828 125 861 25 894375 9275 960625 99375 1026875 1060 1093125 112625 1159375 1192 5 843 75 877 5 911 25 945 978 75 10025 04625 1000 111375 11475 112125 125 859 375 89375 928 125 9625 996875 103125 1065625 1100 1134375 116875 1203125 1237 5 B75 970 345 gau 1015 1080 1085 1120 1155 1190 1225 1280 890 625 92625 961875 99758 1033125 108875 1104375 1140 1175625 121125 1246875 1282 5 906 25 9425 97875 1015 7051 25 10875 112375 160 118625 12325 126875 1305 921875 95875 095625 10325 1069375 110625 1143125 1180 1216875 125375 1290625 1327 5 937 5 975 1012 5 7050 1087 5 1125 1162 5 1237 5 1275 1312 5 1350 953 125 991 258 1029 275 1067 5 1105625 114375 1181 875 1920 12581250 129625 1334375 1372 5 988 75 1007 5 104825 1085 112875 11625 1201 25 1240 197875 13175 135625 1395 984 375 1023 75 1063125 11025 1141 875 118125 1220 625 1260 1299 375 1338 75 1278 125 14175 Page 30 www btxonline com ECM 630 Electroporation System 925 23 125 16 25 69 375 115 625 138 75 161 875 185 231 25 254 375 2775 323 75 346 875 393 125 416 25 439 375 485 625 50875 531 875 948 125 971 25 994 375 1017 5 1040 625 1063 75 1086 875 1170 1133125 1156 25 1179 375 1202 5 1225 625 1248 75 1271 875 1295 1318 125 1341 25 1364 3 5 1387 5 1410 625 1433 75 1456 875 Page 31 950 2375 475 71 25 95
45. 5 2208 75 2244 375 1450 36 25 72 5 10875 145 18125 2175 253 75 290 326 25 362 5 398 75 435 471 25 507 5 543 75 580 616 25 652 5 888 75 725 761 25 7975 833 75 870 906 25 9425 978 75 1015 1051 25 1087 5 1123 75 1160 1196 25 1232 5 1268 75 1305 1341 25 1377 5 1413 75 1450 1486 25 15225 1558 75 1595 1631 25 1667 5 1703 75 1740 1778 25 18125 1848 75 1885 1921 25 1957 5 1993 75 2030 208625 21025 2138 75 2175 2211 25 2247 5 2283 75 1475 1500 1525 36 875 37 5 38 125 73 75 75 76 25 110625 1125 114 375 1475 150 1525 184375 1875 190 625 221 25 2258 22875 258 125 2625 265 875 331 875 337 5 343 125 368 75 375 381 25 405 625 4125 419 375 4425 450 4575 419315 4875 495 825 516 25 525 533 75 553 125 5625 571 875 590 600 510 626 875 6375 648 125 653 75 675 686 25 700 625 725 724375 737 5 750 782 5 774375 7875 800 625 811 25 825 83875 848 125 B625 876 875 885 900 915 921 875 937 5 953 125 958 75 975 991 25 995 625 10125 1029 375 10925 1050 10675 1069 375 1087 5 1 105 625 1106 25 1125 1143 75 1143 125 1162 5 1181 875 1180 1200 1220 1216 875 12375 1258 125 1253 78 1275 129625 1290625 131258 1334375 13275 1350 13725 1364 375 1387 5 1410 625 1401 25 1425 1448 75 1438 1 25 1462 5 1486 875 1476 1500 1525 1511 875 1537 5 1563 1 25 1548 75 1575 1601 25 1585 625 16125 1639 375 15225
46. 5 157 5 161 875 168 25 170 825 175 179 375 183 75 158 125 192 5 196 875 201 25 205 625 210 214 375 218 75 223 125 227 5 231 875 236 25 240 525 245 249 375 253 75 258 125 262 5 266 875 271 25 275 625 www btxonline com 10 170 175 180 185 190 195 210 215 225 230 235 240 245 250 225 5 625 11 25 16 875 225 28 125 3375 39 375 45 50 625 56 25 61 875 675 73 125 78 75 84 375 90 95 625 101 25 106 875 112 5 118 125 12375 129 375 135 140 625 146 25 151 875 157 5 163 125 168 75 174 375 180 185 625 191 25 196 875 202 5 208 125 21375 219 375 225 230 625 238 25 241 875 2475 253 125 258 75 254 375 270 275 625 281 25 286 875 292 5 298 125 303 75 309 375 315 320 625 326 25 331 875 337 5 343 125 348 75 354 375 6 25 25 18 75 3125 375 43 75 50 56 25 625 68 75 75 81 25 87 5 93 75 100 1125 11875 125 131 25 1375 143 75 156 25 1625 158 75 181 25 187 5 193 75 206 25 2125 218 75 231 25 2375 24375 256 25 2625 268 75 275 28125 2875 293 75 306 25 3125 318 75 331 25 337 5 343 75 350 356 25 362 5 36875 375 381 25 387 5 393 75 275 6 875 1375 20 525 275 34 375 4125 48 125 55 61 875 68 75 75 625 82 5 89 375 96 25 103 125 110 116 875 123 75 130 625 137 5 144 375 151 25 158 125 165 17
47. 5 3850 525 3913 75 3976 875 www btxonline com 2550 63 75 127 5 191 25 255 318 75 382 5 446 25 510 573 75 637 5 701 25 765 B2B 75 892 5 956 25 1020 1083 75 1147 5 1211 25 1275 1338 75 1402 5 1466 25 1530 1593 75 1657 5 1721 25 1785 1848 75 1912 5 1976 25 2040 2103 75 2167 5 2231 25 2295 2358 75 2422 5 2486 25 2550 2613 75 2677 5 2741 25 2805 2868 75 2932 5 2996 25 3060 3123 75 3187 5 3251 25 3315 3378 75 3442 5 3506 25 3570 3633 75 3697 5 3761 25 3825 3888 75 3952 5 4018 25 ECM 630 Electroporation System Appendix 630 RC Time Constants Continued Low Voltage ER dn M od e RC 25 00 0 63 1 25 50 00 1 25 2 50 75 00 1 88 3 75 3225 3250 100 00 2 50 5 00 80 625 81 25 125 00 313 625 16125 1625 I 150 00 3 75 7 50 241 875 243 75 175 00 4 38 8 75 3225 325 27 200 00 5 00 10 00 403 125 406 25 225 00 5 63 11 25 483 75 4875 4 250 00 6 25 12 50 364575 568 75 275 00 8 88 13 75 0 300 00 7 50 15 00 325 00 8 13 16 25 KATE se TE 350 00 8 75 17 50 9675 975 375 00 9 38 18 75 5 1048125 105625 400 00 10 00 20 00 E 1128 75 1137 5 25 425 00 10 63 21 25 1209 375 121875 450 00 11 25 22 50 2 1290 1300 475 00 11 88 23 75 1370 625 1381 25 500 00 12 50 25 00 1451 25 1462 5 525 00 13 13 26 25 1531 875 1543 75 550 00 13 75 27 50 16125 1625 137 575 00 14 38 28 75 1693 125 1706 25 600 00 15 00 90 00 D ERE 5 525 00 15 63
48. 5 70 725 775 82 5 85 87 5 92 5 95 97 5 100 102 5 105 107 5 110 112 5 115 117 5 120 122 5 125 127 5 130 132 5 135 137 5 140 142 5 145 147 5 150 152 5 155 157 5 125 3 125 825 9375 12 5 15 625 1875 21875 25 28 125 31 25 34 375 37 5 40 625 43 75 46 875 50 53 125 56 25 59 375 625 85625 68 75 71 875 75 78 125 81 25 84 375 875 90 625 93 75 96 875 100 103 125 106 25 109 975 112 5 115 625 118 75 121 875 125 128 125 131 25 134375 137 5 140 625 143 75 146875 150 153 125 156 25 159 375 1625 165 625 168 75 171 875 175 178 125 181 25 184 375 1875 190 625 19375 196 875 150 3 75 75 11 25 15 1875 225 2625 33 75 37 5 41 25 45 4875 52 5 56 25 63 75 87 5 7125 5 7875 825 86 25 80 9375 975 101 25 105 108 75 112 5 116 25 120 123 75 127 5 131 25 135 13875 142 5 146 25 150 153 75 157 5 16125 185 16875 172 5 176 25 180 183 75 187 5 191 25 195 198 75 202 5 206 25 210 213 75 217 5 221 25 225 22875 232 5 236 25 175 4 375 8 75 13 125 175 21 875 2625 30 625 35 39 375 43 75 48 125 52 5 56 875 61 25 65 625 70 74 375 7875 83 125 8 5 01 875 95 25 100 625 105 109 375 11375 118 125 122 5 196 875 131 25 135 525 140 144 375 14875 153 12
49. 5 200 206 25 2125 218 75 225 171 875 178 75 185 625 192 5 199 375 206 25 213 125 22 226 875 23375 240 625 2475 1875 195 202 5 210 2175 225 232 5 240 2475 255 262 5 279 203125 21125 2192375 2275 23562 24375 251 875 260 268125 27625 284375 292 5 218 75 2275 236 25 245 253 75 262 5 271 25 280 28875 297 5 306 25 315 5 234 375 243 75 253 125 2625 271 875 281 25 290 625 300 308 375 31875 328 125 337 5 265 625 27625 286875 2975 308125 31875 329375 340 350 625 36125 371 875 3825 281 25 292 5 303 75 375 326 25 337 5 348 75 360 371 25 382 5 393 75 405 296 875 308 75 320 625 332 5 344 375 356 25 368 125 380 391 875 403 75 415 625 4275 3125 325 3375 330 362 5 375 387 5 400 4125 425 4375 450 328 125 341 25 354375 367 5 380 625 39375 406 875 420 433 125 446 25 458 375 472 5 343 75 3575 371 25 385 398 75 4125 426 25 440 453 75 467 5 481 25 495 359 375 37375 388125 40258 416875 43125 445 625 480 474375 48875 503125 8175 375 390 405 420 435 459 465 480 495 510 525 540 390 625 40625 421 875 4375 453125 46875 484375 500 515625 53125 546875 5525 400 25 4225 438 75 455 471 25 487 5 503 75 529 536 25 552 5 56875 585 421 875 43875 455625 47258 489375 50625 3523 125 540 556875 57375 590625 607 5 437 5 455 472 5 490 507 5 525 542 5 580 577 5 595 6125 830 453 125 47125 489375 5075 525625 54275 561875 580 598125 81625 634575 652 5 468 75 4875 50625 525 54375 5625 58125 800 851875 837 5 858 25 875 484 375 503 75 523125 5425 3561875 581 25 600625 820 63
50. 5 625 2005 2244 375 2283 75 2323 125 2362 5 2401 875 2441 26 2480 625 ECM 630 Electroporation System 640 680 720 760 800 840 8B0 920 960 1000 1040 1080 1120 1160 1200 1240 1280 1320 1350 1400 1440 1480 1520 1560 1600 1640 1680 1720 1760 1800 1840 1880 1920 1960 2000 2040 2080 2120 2160 2200 2240 2280 2320 2360 2400 2440 2480 2520 Capacitor uF 1625 1650 40 625 41 25 81 25 82 5 191875 12375 182 5 165 203 125 206 25 243 75 247 5 284 375 288 75 325 330 365 625 37125 405 25 412 5 446 875 453 75 487 5 495 528 125 536 25 568 75 577 5 609 375 61875 650 660 690 625 701 25 731 25 742 5 771 875 783 75 812 5 825 853 125 866 25 89375 907 5 934 375 94875 975 990 1015 6825 103125 105625 10725 1098 875 111375 1137 5 1155 1178 125 119625 1218 75 1237 5 1259 375 127875 1300 1320 1340 625 1361 25 1381 25 1402 5 1421 875 1443 75 1462 5 1485 150325 152625 154375 1567 5 1584375 160875 1625 1850 1885825 169125 170825 1732 5 1746 875 177375 1787 5 1815 1828 125 1856 25 1868 75 1897 5 1909 375 193875 1950 1980 1890 625 2021 25 2031 25 2062 5 2071 875 210375 2112 5 2145 2153 125 2186 25 219375 2227 5 2234375 206875 2275 2310 2315 625 2351 25 235625 23825 2396 875 243375 2437 5 2475 2478 125 2516 25 2518 75 2557 5 2559375 250875 Page 33 1675 41 875 8375 125 625 167 5 209375 251 25 293 125
51. 60 525 3112 5 3164 375 3216 25 3288 125 2100 52 5 105 157 5 210 252 5 315 357 5 472 5 525 577 5 682 5 2125 2150 2175 53 125 53 75 54 375 106 25 1075 10875 159 375 18125 163125 212 5 215 2175 265 625 26875 271 875 318 75 3225 326 25 371 875 376 25 380 625 425 430 435 478 125 483 75 489 375 531 25 537 5 543 175 584 375 591 25 598 1 25 637 5 645 5525 690 525 698 75 706 875 743 75 752 5 751 25 796 875 806 25 815 625 850 850 870 903 125 91 3 75 924 375 958 25 9675 97875 1009 375 102125 1033 125 1082 5 33075 10875 1115 525 1128 75 1141 875 1168 75 1182 5 1196 25 1221 875 1236 25 1250 625 1275 1290 1305 1328 125 134375 1359 375 138125 13975 141375 1434 375 145125 1468 125 1487 5 1505 1522 5 1540 525 1558 75 1576 875 1593 75 1612 5 1631 25 1646 875 166625 1685625 1700 1720 1740 1753 125 177375 1794375 1808 25 18275 184875 1859 375 1881 25 1903 125 1912 5 1935 19575 1965 525 1988 75 2011 875 201875 20425 206625 2071 875 209625 2120 625 2125 2150 2175 2178 125 2203 15 2229375 223125 22575 228375 2284 375 2311 25 2338 125 2337 5 2365 2392 5 2390 525 2418 75 2446 875 244375 24725 2501 25 2496 875 252625 2555625 2550 2580 2610 2803195 263375 2664375 2656 25 26875 271875 2709 375 2741 25 2773 125 2752 5 2795 2827 5 2815 625 2448 75 2881 875 2868 75 29025 203625 2921 875 2956 25 2990 625 2975 3010 3045 3028 125 3063 75 3099375 3081 25 31175 3153 75
52. 75 810 860 625 91125 961 875 1012 5 1063 125 1113 75 1164 375 1215 1285 625 1316 25 1368 875 1417 5 1468 125 1518 75 1569 375 1820 1670 625 1721 25 1771 875 1822 5 1873 125 1923 75 1974 375 2025 2075 625 2126 25 2176 875 2227 5 2278 125 2328 75 2379 375 2430 2480 625 2531 25 2581 875 2632 5 2683 125 2733 75 2784375 2835 2885 625 2936 25 2586 875 3037 5 3088 125 3138 75 3189 375 2050 51 25 102 5 153 75 205 256 25 307 5 358 75 410 461 25 512 5 563 75 815 666 25 717 5 768 75 820 871 25 922 5 973 75 1025 1076 25 1127 5 1178 75 1230 1281 25 1332 5 1383 75 1435 1486 25 1537 5 1588 75 1840 1691 25 17425 1793 75 1845 1896 25 1947 5 1998 75 2050 2101 25 21525 2203 75 2255 2306 25 2357 5 2408 75 2460 2511 25 2562 5 2613 75 2665 2716 25 2767 5 2818 75 2870 2921 25 29725 3023 75 3075 3126 25 3177 5 3228 75 2075 51 875 103 75 155 825 207 5 259 375 31125 363 125 415 466 875 518 75 570 825 622 5 674 375 726 25 778 125 830 881 875 933 75 985 525 1037 5 1089 375 1141 25 1193 125 1245 1296 875 1348 75 1400 525 1452 5 1504 375 1556 25 1608 125 1660 1711 875 1763 75 1815 525 1867 5 1918 375 1971 25 2023 125 2075 2128 875 2178 75 2230 525 2282 5 2334 375 2386 26 2438 125 2490 2541 875 2593 75 2645 625 2697 5 2749 375 2801 25 2853 125 2905 2956 875 3008 75 30
53. 875 4134 375 2860 6625 1325 198 75 265 381 25 3975 46375 530 596 25 662 5 72875 795 861 25 9275 99375 1060 1126 25 11925 1258 75 1325 139125 14575 152375 1590 165625 17225 1788 75 1855 1921 25 19875 2053 75 2120 2186 25 2252 5 2318 76 2385 245125 2517 5 258375 2650 271625 27825 284875 __ 2915 2981 25 30475 3113 75 8180 3248 25 33125 3378 75 3445 3511 25 3577 5 364375 3710 3776 25 38425 3908 75 3975 4041 25 41075 417375 2675 66 875 133 75 200 625 267 5 334 375 40125 468 125 535 601 875 668 75 735 625 8025 869 375 93825 1003 125 1070 1136 875 1203 75 1270 625 1337 5 1404 375 147125 1538425 1605 1671 875 173875 1805 625 18 25 1939 375 200625 2073 125 2140 2206 875 227375 2340 625 24075 2474 375 2541 25 2608 125 2675 2741 875 2808 75 2875 625 28425 3009 375 3076 25 3143 125 3210 3278 875 3343 75 3410 625 3477 5 3544 375 3611 25 3678 125 3 45 3811 875 3878 75 3945 625 4012 5 4079 375 4146 25 4213 125 2700 675 2025 270 337 5 472 5 607 5 675 7426 810 8775 945 1012 5 1080 1147 5 1215 1282 5 1350 14175 1485 15525 1620 1687 5 1755 1822 5 1890 19575 2025 2092 5 2160 2027 5 2295 23825 2430 2497 5 2632 5 2700 2767 5 2835 28025 2970 3097 5 3105 3172 5 3240 3307 5 3975 3442 5 3510 3577 5 3645 3712 5 3780 3847 5 3915 3082 5 4050 4117 5 4185 4252 5
54. 875 853125 6875 721875 758 25 790 825 825 420 455 490 525 580 595 630 865 700 735 770 805 840 4275 468425 49875 534375 970 605625 641 25 670 875 7125 748425 783 75 8619 375 855 435 471 25 5075 54375 580 61625 5525 68875 725 76125 7975 83375 870 4425 479 378 51825 553 125 590 626875 868375 700825 7375 774375 811 25 848 125 885 450 487 5 525 562 5 600 837 5 675 712 5 750 787 5 825 862 5 900 457 5 495 625 53375 571 875 610 648 125 686 26 724 375 762 5 800 625 838 75 875 875 915 465 50375 542 5 581 25 620 658 75 6975 73625 775 81375 8525 89125 930 4725 511 875 55125 590 625 630 659 375 70875 748125 7875 826875 865 25 805 625 945 Page 29 www btxonline com ECM 630 Electroporation System Appendix 630 RC Time Constants Continued Low Voltage Mode RC 825 850 575 700 725 750 775 800 825 850 875 900 15625 16 28 16 875 1775 18 125 1878 19 375 20 20 625 2125 21 875 225 31 25 325 33 75 35 3625 375 3875 40 41 25 425 43 15 45 46 875 48 75 50 625 525 54 375 5625 58 125 89 51 875 6375 85 625 575 625 65 675 70 72 5 75 775 80 82 5 85 875 90 78 125 81 25 84 375 875 90 625 9375 96 875 100 103125 16625 109375 1125 93 75 875 101 25 105 108 75 112 5 116 25 120 123 75 127 5 131 25 135 109 375 113 75 118 125 122 5 126 875 13125 135 625 140 144 375 148 75 153 125 157 5 125 130 135 140 145 150 155 160 165 170 175 180 140 625 14625 151875 1575 163125 16875 474375 180 185625 19125 196875 2025 156 25 1625 168 75 175 181 25 1875 193 7
55. 9 and are currently exempt from the RoHS Directive We will continue to monitor the application of the RoHS Directive to its products and will comply with any changes as they apply Do Not Dispose Product with Municipal Waste Special Collection Disposal Required ECM 630 Electroporation System Table of Contents General Information Serial gt ese l IURI DIEM TIMERE DITE 2 C I 2 eec 25 3 4 Repair Facilities and Parts 4 General Safety Summary 5 6 Electrical amp Technical Specifications 7 General Specifications 8 Introduction ECM 630 Features 9 Operation Getting Started 10 Quick Start u u EE sani n k u ET 11 C DI eT 11 seyan s 11 Instrument Controls 12 17 Operating BASICS 18 19 Advanced Operation Programming 20 Electroporation 21 Applications uwa rrr rrr 2ox2o 22 23 Appendix A 630 Electrode Operation Ranges 24 26 Appendix B 630 RC Time Constants
56. 9375 85875 678125 097 5 500 520 540 560 580 909 520 540 560 680 700 720 515 625 536 25 555 875 577 5 598 125 61875 639 375 560 69 625 701 25 721 875 742 5 531 25 5525 573 75 595 616 25 637 5 658 75 701 25 7225 743 75 765 546 875 568 75 3590 625 6125 634 375 65625 878 125 700 721 875 74375 785 625 787 5 5625 585 607 5 830 652 5 875 697 5 720 742 5 785 787 5 810 578125 80125 6243 5 6475 670625 69375 716875 40 753125 78625 809375 8325 593 75 6175 641 25 965 688 75 7125 736 25 760 783 75 807 5 831 25 855 509 375 633 75 658 125 6825 706 875 73125 755625 780 804375 82875 853425 877 5 525 550 575 700 725 750 775 800 825 850 875 200 640625 66625 891 975 7175 743125 78875 794375 820 845625 871256 896875 9225 656 25 8825 708 75 738 761 25 787 5 813 75 840 866 25 892 5 918 75 945 871875 89675 725825 7525 779375 80625 833125 860 886875 91375 840625 957 5 8875 715 7425 770 7975 825 852 5 9075 sas 9525 990 703 125 731 25 759 375 787 5 815 625 843 75 871875 900 928 125 956 25 984 375 10125 718 75 747 5 776 25 805 833 75 862 5 891 25 920 948 75 977 5 1006 25 1035 734 375 763 75 793125 8225 851 875 88125 910 625 940 959 375 99875 1028 125 1057 5 750 780 810 840 870 909 330 980 990 1020 1050 1080 765 625 796 25 826 875 857 5 88 125 918 75 949 375 980 1010 625 104125 1071 875 1192 5 7h 25 8125 B43 75 875 906 25 9375 968 75 1000 103125 10625 109375 1125 796 875 82875 860625 8925 924375 95625 988 125 1020 1051875 108
57. ard At pulse lengths at or above 1 to 2 ms beeps should be heard 3 Following the pulsing status message the Pulse Delivered Screen is displayed 4 Press the encoder or the Pulse switch to return to Set Parameters screen Pulsing Aborted During Charging 1 If the start switch is activated a second time prior to delivery of a pulse the pulsing sequence is aborted and the Pulsing Aborted status message is displayed 2 Press the encoder or the pulse switch once to get back to the Set Parameters screen Page 16 www btxonline com ECM 630 Electroporation System Quick Start Continued Pulse Delivered Screen 1 The Pulse Delivered Screen is displayed following the delivery of a pulse and is indicated by the display Pulse Delivered on the top line of the screen 2 Press the knob to return to the Set Parameters Screen 3 The Pulse Delivered screen displays the monitored peak Voltage and Time Constant on page 1 and the programmed Resistance and Capacitance on page 2 Please note that a pulse will be delivered and monitored even if there is no output so do not use this feature to verify that a pulse was delivered to your sample 4 Always verify that your sample is connected to the HV output Voltage 1 Voltage indicates the peak voltage delivered The voltage delivered is affected by the external load Sample 2 Using heavy loads low resistivity will result in a slightly lower delivered volta
58. be overwritten once a program is loaded A default program is used to initialize the system Program 1 is used to automatically store changes in parameters set each time the system is pulsed Programs 2 and 3 can be used to set and save experimental parameters for instantaneous use To save the current set parameters as a program push the parameter control knob to move the arrow to the left of the program number Rotate the knob to designate the appropriate program number then push and hold the knob in releasing after a confirmation screen is displayed The confirmation screen will read Saved Current Parameters to Set N where N is a number from 1 to 3 Push the parameter control knob again to return to the Set Parameters screen www btxonline com ECM 630 Electroporation System Quick Start Continued View 1 To view the appropriate program parameters push the parameter control knob to move the arrow to the left of the program number 2 Select the appropriate program number by rotating the parameter control knob then hold the knob in releasing after a new screen is displayed 3 The new screen will display the parameters currently stored under that program number 4 Push the parameter control knob again to return to the Set Parameters screen Load 1 To load a saved program push the parameter control knob to move the arrow to the left of the program number Please note that in addition t
59. beep when toggling from one page to another Voltage 1 Voltage indicates the set voltage in volts 2 The low voltage mode range is 10 V to 500 V in 1V increments and the high voltage mode range is 50 V to 2500 V in 5 V increments 3 The mode is indicated by either LV for low voltage or HV for high voltage after the voltage value 4 Abeepis heard when transitioning between and HV modes Page 12 www btxonline com ECM 630 Electroporation System Quick Start Continued Resistor 1 2 Resistor indicates the set resistance in The resistance range is 25 O to 1575 O in 25 O increments In the mode there is an additional resistor selection of None This indicates that there is no timing resistor in the pulsing circuit thus mimicking settings for competitive electroporators in which there is either no resistor to select or the resistance is termed unlimited Capacitor 1 2 Capacitor indicates the set capacitance microfarads pF In the LV mode the capacitance range is 25 to 3275 pF 25 pF increments In the HV mode the capacitance choice is 25 pF and 50 pF 4 The capacitance and resistance control the time constant Please refer to Appendix B for explanation and time constant charts Advanced Features 1 Save Page 13 The second Set Parameters screen enables the user to save view and load up to three programs Please note that current parameters will
60. chnical Terms continued Protoplasts The plant cell proper with the cellulose cell wall removed Relaxation Time The time a system requires to reach equilibrium Transfection The introduction of nucleic acids into animal cells Stable transfections result in integration of nucleic acids into host chromosomes and the inheritance of associated traits in progeny cells Transient transfections result in temporary expression of exogenous nucleic acids Transformation The introduction of nucleic acids into microorganisms and plant cells Turgor Pressure The pressure in capillaries Page 44 www btxonline com ECM 630 Electroporation System Appendix H Electroporation Pulse Generator Compatibility Certain components of BTX Electroporation are compatible with components of competitive systems Please contact BTX Technical Support for details Page 45 www btxonline com ECM 630 Electroporation System Appendix I Recommended Reading Eberhard Neumann Editor Electroporation and Electrofusion in Cell Biology Plenum Publishing Corporation 1989 Michael Kriegler Gene Transfer and Expression A Laboratory Manual Stockton Press 199 Donald Chang Editor in Chief Guide to Electroporation and Electrofusion Academic Press 1992 Jac A Nickoloff Editor Electroporation Protocols for Microorganisms in Methods in Molecular Biology Vol 47 Humana Press 1995 Jac A Nickoloff Editor Animal Cell Electroporation and E
61. ently high field strength may increase the membrane potential past a critical point leading to the breakdown of the membrane Dielectrophoresis A consequence of cells being exposed to an inhomogeneous or divergent electric field resulting in their movement toward electrodes and subsequent alignment or pearl chain formation Electroinsertion The use of electroporation to insert molecules into lipid bilayer membranes Electropermeabilization The use of electroporation to make cells protoplasts or liposomes permeable to ions and small molecules in their extracellular environment Electroporation The application of high electric field pulses of short duration to create temporary pores holes in the membranes of cells Hydrostatic Pressure The pressure in liquids at rest Lipid Bilayer An assembly of lipid and protein molecules held together by non covalent interactions All biological membranes share this common structure Osmotic Pressure The applied pressure required to prevent the flow of solvents of different concentration across a semipermeable membrane Pore A small mostly transient opening in a cell wall caused by the application of a brief high electric field pulse Pressure Gradient The difference in pressure between two points in a medium Page 43 Disposable Electroporation Cuvettes Plus www btxonline com ECM 630 Electroporation System Appendix G Glossary of Biological amp Te
62. es of transfectant molecule Is the DNA well purified Try reducing the voltage or increase sample volume until arcing is no longer a problem Low or no transfection efficiency or incorporation Verify physical biological and chemical parameters Verify delivery of the pulse and pulse parameters Is the voltage correct Chamber gap Pulse length or appropriate instrument settings Number of pulses If so follow Optimization Guidelines outlined in Appendix A Low viability Verify physical biological and chemical parameters Are the voltage chamber gap pulse length time constant pulse number and other instrument settings correct If so reduce voltage pulse length or number of pulses and re optimize protocol to improve viability as outlined in Appendix A Page 41 www btxonline com ECM 630 Electroporation System Appendix F Glossary of Electrical Terms Amplitude The instantaneous value of current or voltage in amperes or volts Capacitor A device that stores electric energy in the form of an internal electric field Energy is delivered when a current flows out of a capacitor The current normally follows an exponential curve Dielectric A material that has a high resistivity and can store energy in the form of an electric field Direct Current DC Current whose amplitude is constant with time Direct currents are used to form temporary pores in bi lipid membranes Cells may fuse when pores in the membra
63. ge This effect is normal and will be more pronounced with small capacitor settings Time Constant Time Constant indicates the exponential decay time constant 1 e in ms ms s Page 17 www btxonline com ECM 630 Electroporation System Operating Basics Use with Safety Stand 630B and BTX Disposable Electroporation Cuvettes Plus 1 Insert the safety stand banana plugs into the HV Output on the front panel of the ECM 630 2 Adjust the distance between the metal contacts using the black roller for your cuvette See 630 B instruction sheet 3 Press the green power switch to initialize the 630 4 Rotate parameter control knob to move arrow to voltage and push to select Rotate knob to adjust voltage Push again to select that voltage 5 Rotate parameter control knob to move arrow to Resistor and push to select Rotate knob to adjust resistance Push again to select that resistance 6 Rotate parameter control knob to move arrow to Capacitor and push to select Rotate knob to adjust the capacitance Push again to select that capacitance 7 Prepare sample pipette into the appropriate BTX Disposable Cuvettes Plus place the cuvette in the 630B Safety Stand and secure the safety cover 8 Press the Start button The 630 will charge and then deliver the electroporation pulse while beeping 9 Process sample Do not forget to record appropriate parameters as displayed on the Pulse Delivered sc
64. he product Consult the product manual for further ratings information before making connections to the product DO NOT OPERATE WITHOUT COVERS Do not operate this product with covers or panels removed Use Proper Fuse Use only the fuse type and rating specified for this product AVOID EXPOSURE TO CIRCUITRY Do not touch exposed connections and components when power is present DO NOT OPERATE IN LOW IMPEDANCE Sample Load or Sample If the electroporation samples have an impedance of less than 20 in and 40 in HV the samples may arc and result in sample loss and potential damage to unit DO NOT OPERATE WITH SUSPECTED FAILURES If you suspect there is damage to this product have it inspected by qualified BTX service personnel PROVIDE PROPER VENTILATION Refer to installation instructions for details on installing the product to ensure proper ventilation Page 5 www btxonline com ECM 630 Electroporation System General Safety Summary Continued DO NOT OPERATE IN WET DAMP CONDITIONS DO NOT OPERATE IN AN EXPLOSIVE ATMOSPHERE KEEP PRODUCT SURFACES CLEAN AND DRY Should you have any safety concerns immediately contact BTX Technical Services 1 800 272 2775 Safety Terms and Symbols Terms that appear in this manual WARNING Warning statements identify conditions or practices that could result in injury or loss of life CAUTION Caution statements identify conditions or practices that could result in da
65. he following items com plete a typical system order Model 630B Electroporation Safety Stand 1 Model 660 Cuvette Rack 1 Model 610 BTX Cuvettes Plus 10 Model 620 BTX Cuvettes Plus 10 Model 640 BTX Cuvettes Plus 10 If you have ordered alternative or different items please verify their receipt Page 15 www btxonline com ECM 630 Electroporation System Quick Start Continued The value of the timing capacitance Time constant t RC Resistor The value of the timing resistance Time constant t RC Save Save current setup parameters to nonvolatile memory View Show the setup parameters stored in nonvolatile memory Load Load setup parameters from memory to use Status Messages Following the initiation of a pulsing sequence various status messages are displayed The following status messages may be observed Charging Pulsing and Pulse Aborted During Charging Charging As soon as the start switch is pressed the Charging status message is displayed The capacitors in the ECM 630 are charged during the duration of this message Pulsing 1 Once the capacitors have reached the preset voltage level they are discharged and the Pulsing status message is displayed for the duration of the pulse 2 Aclick or a beep will be heard during the delivery of the pulse The sound that is heard is a function of the pulse length At pulse lengths less than 1 to 2 ms clicks will likely be he
66. i The plate size should be selected according to the electric field desired the desired volume of the sample and the capabilities of the electroporator 2 Load Plate with Cells 3 Each column should be loaded with the same number of samples if possible i Load unused wells in a column with the same sample media at the same volume 3 Connect the Model HT 100 to the color coded banana cable taking care to match the color of the cable to the color ring around the connector and then plug the banana cable into the voltage output of the electroporator again matching the color polarity 4 Place plate HT 96 or the adapter frame containing plate HT 25 onto the plate handler so the plate matches the nest plate outline on the handler 5 Close HT 100 Plate Handler lid firmly to latch The front panel latch pops out when securely closed When lid is closed correctly the LED for column 1 should flash 6 Select appropriate column 1 12 to begin electroporation by using the column adjust buttons 3 The unit will default to column 1 when lid is closed b Pressing and holding the button allows the unit to rapidly advance through columns after a short pause Page 52 www btxonline com ECM 630 Electroporation System Appendix M Plate Handler Continued 7 Configure the electroporator with appropriate settings for voltage resistance and time capacitance Setthe electroporator for the total number of pulses
67. lectrofusion Protocols in Methods in Molecular Biology Vol 48 Humana Press 1995 Jac A Nickoloff Editor Plant Cell Electroporation and Electrofusion Protocols in Methods in Molecular Biology Vol 55 Humana Press 1995 For further references regarding specific applications and optimiza tion please contact Technical Support BTX Division of Harvard Apparatus 84 October Hill Road Hollistion MA 01746 Phone 1 508 893 8999 Toll Free 1 800 272 2775 Fax 1 508 429 5732 Email techsupport btx harvardapparatus com Website www btxonline com Page 46 www btxonline com ECM 630 Electroporation System Appendix J Accessories and Replacement Parts Catalog No 45 0001 45 0051 45 0207 45 0124 45 0125 45 0126 45 0400 45 0450 45 0452 45 0463 45 0462 45 0465 45 0466 45 0467 45 0059 Page 47 Model 6300 630 630B 610 620 640 HT100 HT P96 2 HT P96 4 HT P25 P4 HT P25 4 HT P25 2 25 2 30007 Description 630 Electroporation System ECM 630 Electroporator only Electroporation Safety Stand Disposable Electroporation Cuvettes Plus 1mm 50 bag Disposable Electroporation Cuvettes Plus 2 mm 50 per bag Disposable Electroporation Cuvettes Plus 4 mm 50 per bag Manual 96 Well Plate H ndler Disposable 96 Well Plate 2mm gap 96 Disposable 4mm 25 Well Disposable 4mm pkg 6 25 Well Disposable
68. mage to these products or other property Symbols that may appear on the products A Danger Attention Protective Functional High Refer to Earth Ground Voltage Manual Terminal Page 6 www btxonline com ECM 630 Electroporation System Electrical amp Technical Specifications Standard Capabilities Display Power Source Voltage Power Fusing 20 character by 4 line liquid crystal Display LED backlit 100 to 240 Vac 50 to 60 Hz CAT II 500 W Pulsed 50 W Idle 2 5 T rating 250 V Environmental Characteristics Intended Use Operating Temperature Cooling Relative Humidity Altitude Mechanical Characteristics Footprint Weight Controls Page 7 Indoor use only 10 C to 40 C Convection throush metal case 6096 2 000 m operating 12 5 x 12 25 x 5 5 W x Dx IH 13 6 Ibs 6 2 kg Single rotary encoder with integrated push button www btxonline com ECM 630 Electroporation System General Specifications CAUTION Certifications and Compliances FOR RESEARCH USE ONLY Overvoltage Category NOT FOR CLINICAL CAT 111 Products in this Category Distribution level mains USE ON PATIENTS fixed installation Local level mains applications portable equipment CAT I Signal levels in special equipment or parts of equipment telecommunications electronics Meets requirements of Directive 89 336 EEC for Electromagnetic Compatibili
69. nd release Now rotate to select which program number the chosen settings see Preset Parameters above should be saved under Program 1 is reserved for current parameters in active use and cannot be used for pre set storage Push and hold knob in releasing only after a new screen reading Saved Current Parameters to Set N is displayed N refers to program number designated in step 2 Push the parameter control knob again to return to the Set Parameter Screen From the Set Parameter Screen use the parameter control knob to rotate until the load function has been reached Push the knob in and release Now rotate to select the appropriate program number Push and hold knob in releasing only after a new screen reading Loaded Set N to Current Parameters is displayed Push the parameter control knob again to return to the Set Parameter Screen www btxonline com ECM 630 Electroporation System Electroporation Electroporation is the application of controlled pulsed electric fields to biological systems If the biological system contains a lipid bilayer such as is the case if the system is a suspension of cells or liposomes the pulsed electric field may overcome the field potential of the lipid bilayer resulting in a reversible breakdown of the bilayer and a resulting formation of temporal pores in the membrane The pores formed are of the order of 40 to 120 nm Most pores reseal within a few seconds after allowing the
70. nes of two juxtaposed cells reseal after a DC application Divergence The deviation of electric field lines from a parallel homogeneous condition A highly divergent field has field lines that rapidly change amplitude or strength and direction in the area of interest Electric Field The electric potential difference between two points divided by the distance separating those points Expressed in volt cm Electric Field Force The mechanical force acting on any electric charge when placed in an electric field Exponential Decay Non linear waveform typical of capacitor charge and discharge currents and voltages The exponential decay waveform is characterized by its time constant the time it takes the voltage to decay to 1 e of the peak voltage Field Strength See Electric Field Frequency The number of times an oscillation goes through a complete cycle in one second The unit is either cycle sec or Hz Homogenous Electric Field An electric field where the direction and strength of the field lines are constant Page 42 www btxonline com ECM 630 Electroporation System Appendix G Glossary of Biological amp Technical Terms Chambers Electroporation and Electrofusion Chambers are the devices used to hold the cells molecules to be fused transfected Dielectric Breakdown The reversible breakdown of lipid bilayer membranes as a result of the application of a DC electroporation pulse Suffici
71. nipulation ECM lies in selecting appropriate ECM systems capable of delivering the pulses suitable for the cell being electromanipulated One or several pulses of the appropriate field strength pulse length and wave shape may be required for this purpose The key to success with electroporation based technologies involves a proper combination of biological physical chemical and pulse parameters In general cells must be in mid logarithmic growth for optimal electroporation Various temperature regimens have been described It has been shown that a variety of chemical techniques may increase electroporation efficiencies including addition of EDTA DMSO intracellular salts and serum before or after the pulse Optimizing protocols abound Analysis of these optimization regimens has lead to proposals of universal protocols involving very limited optimization over a narrow range Electroporation 1 Vary the voltage in order to vary the field strength kv cm keeping other parameters constant Assay sample for both viability and endpoint Plot the field strength versus both viability and endpoint and extrapolate the optimal field strength voltage divided by gap size and voltage 2 Vary the capacitance resistance sample volume in order to vary the pulse length time constant for exponential decay instruments Directly vary square wave instrument pulse length Assay sample for both viability and endpoint Plot the pulse length versus bo
72. o misuse neglect accident or abuse b repaired or altered by anyone other than BTX HARVARD APPARATUS without BTX HARVARD APPARATUS express and prior approval c used in violation of instructions furnished by BTX HARVARD APPARATUS This warranty extends only to the original customer purchaser Failure to use the Enhancer 3000 High Voltage probe to connect a BTX Generator to an external digital oscilloscope for monitoring will result in voiding your warranty connecting directly to the external monitoring equipment or modified monitoring setup will damage the Generator IN NO EVENT SHALL BTX HARVARD APPARATUS BE LIABLE FOR INCIDENTAL OR CONSEQUENTIAL DAMAGES Some states do not allow exclusion or limitation of incidental or consequential damages so the above limitation or exclusion may not apply to you THERE ARE NO IMPLIED WARRANTIES OF MERCHANTABILITY OR FITNESS FOR A PARTICULAR USE OR OF ANY OTHER NATURE Some states do not allow this limitation on an implied warranty so the above limitation may not apply to you Without limiting the generality of the foregoing BTX HARVARD APPARATUS shall not be liable for any claims of any kind whatsoever as to the equipment delivered or for non delivery of equipment and whether or not based on negligence Warranty is void if the 630 is changed in any way from its original factory design or if repairs are attempted without written authorization by BTX HARVARD APPARATUS War
73. o three available programs there is also the default program as outlined in the Initializing section Rotate the knob to change the program number 3 Push and hold the knob in releasing after a confirmation screen is displayed The confirmation screen will read Loaded Set N to Current Parameters where N is a number from 1 to 3 4 Push the parameter control knob again to return to the Set Parameters screen On Line Help 1 In the Set Parameter Mode an On Line Help function is available On Line Help provides a definition for all set parameters and advanced functions 2 use On Line Help rotate the parameter control knob so that the arrow is to the left of the parameter or feature of interest not the value of the parameter 3 Push the parameter control knob in and hold until the definition is displayed 4 Push a second time to return to the Set Parameters screen Mode Low voltage or high voltage range mode display only this function is controlled by the voltage parameter Voltage Peak amplitude of the output pulse Page 14 www btxonline com ECM 630 Electroporation System Operation Getting Started Carefully open the box containing the ECM 630 Electroporation System Verify receipt of the following items 630 Pulse Generator 1 Power Cord 1 Model 630E Electronic Manual 1 BTX Electroporation Systems may be customized with the addition of various electrodes and accessories T
74. oporation can be characterized by waveform BTX exponential decay waveform generators such as the ECM 399 and ECM 630 deliver an exponentially decaying pulse The length of such a discharge waveform is commonly characterized by the time required for the initial voltage to decay to 1 e roughly 1 3 of the initial value To achieve a desired pulse length appropriate resistance and capacitance must be selected on the instrument Voltage may be Exponential Decay Wave directly set on the instrument Page 21 www btxonline com ECM 630 Electroporation System Applications Electroporation Bacteria and Yeast Electroporation The most common application is transformation Field strength and pulse length are critical parameters for reporting optimization and troubleshooting bacterial and yeast applications Mammalian Cell Electroporation Electroporation has been used successfully to introduce many different molecule types into cells Most commonly electroporation is used for the processes of transfection in which nucleic acid DNA and RNA is introduced into cells Electroporation can be used to deliver oligonucleotides into cells for gene silencing or anti sense applications It can be used to deliver proteins into cells even large enzymes such as restriction enzymes and antibodies for various purposes Peptides have also been elec troincorporated Smaller molecules have been incorporated into cells and liposomes such as
75. ranty is void if parts connections or cell fusion chambers not manufactured by BTX HARVARD APPARATUS are used with the 630 Page 2 www btxonline com ECM 630 Electroporation System General Information continued If defect arises within the warranty period promptly contact Harvard Apparatus 84 October Hill Road Building 7 Holliston Massachusetts USA 01746 1388 using our toll free number 1 800 272 2775 US Only or 508 893 8999 E mail techsupport bt lt harvardapparatus com Goods will not be accepted for return unless an RMA Returned Materials Authorization number has been issued by our customer service department The customer is responsible for shipping charges Please allow a reason able period of time for completion of repairs replacement and return If the unit is replaced the replacement unit is covered only for the remainder of the original warranty period dating from the purchase of the original device This warranty gives you specific rights and you may also have other rights which vary from state to state Service All service under the warranty will be made at the HARVARD APPARATUS Holliston Massachusetts facilities or an authorized service site Owner will ship instrument prepaid to Holliston Massachusetts USA or the service site HARVARD APPARATUS will return the instrument after servicing freight prepaid to owner s address Obtaining Service Service D
76. reens for documentation purposes 10 To return to ready mode press the Pulse button 11 To abort a pulse before delivery press the Pulse button during the charging mode Use with Alternative Electrode Chambers and Applications The ECM 630 may be used to power all electroporation cuvettes and electrodes Follow the above instructions in conjunction with instructions provided for the specific electrode Refer to Appendix A for graphical representation of operating ranges of various chambers and applicators with the 630 Reproducing 600 Protocols The ECM 630 will reproduce all 600 protocols 1 Set the voltage as outlined in 600 protocol 2 Calculate the RC time constant without factoring in the external load by multiplying the ECM 600 protocol resistance and capacitance values and dividing by 1000 to give a result in msec Page 18 630B 45 0207 www btxonline com ECM 630 Electroporation System Operating Basics Continued 3 Refer to Appendix B Find the capacitance setting from step 2 and read down the column to find a RC time constant that closely matches Extrapolate the new 630 resistance setting 4 Set the capacitance from the 600 protocol and the new resistance extrapolated from Appendix B 5 Follow experimental procedures while optimizing the voltage as outlined in Appendix C Reproducing Competitive System Protocols
77. stem Appendix 630 Electrode Operation Ranges Continued Operating Range for 620 Water HV 2 gap cuvette 50 to 2500 V 2625 660 usec to 38 msec 2500 pr 5 2375 2250 2125 2000 0 88 5 Length ms 2620 200 Figure 6 Operating Range for Model 620 H 0 HV Page 26 www btxonline com ECM 630 Electroporation System Appendix 630 RC Time Constants SEE PAGES 28 38 Page 27 www btxonline com ECM 630 Electroporation System Appendix 630 RC Time Constants Continued Low Voltage Mode RC 25 00 50 00 75 00 100 00 125 00 150 00 175 00 200 00 225 00 250 00 275 00 300 00 325 00 350 00 375 00 400 00 425 00 450 00 475 00 500 00 525 00 550 00 575 00 800 00 525 00 650 00 875 00 70000 725 00 750 00 775 00 800 00 825 00 850 00 875 00 800 00 925 00 950 00 975 00 1000 00 1025 00 1050 00 1075 00 1109 00 1125 00 1150 00 1175 00 1209 00 1225 00 1250 00 1275 00 1300 00 1325 00 1350 00 1375 00 1400 00 1425 00 1450 00 1475 00 1500 00 1525 00 1550 00 1575 00 Resistor ohm Page 28 0 025 0 05 0 075 0 1 0 125 0 15 0 175 0 2 0 225 0 25 0 275 0 3 0325 0 35 0375 04 0 425 0 45 0475 05 0525 0 55 0575 06 0625 0 55 0675 07 0725 0 75 0 775 0 8 0 825 085 0 875 0 9 0925 0 95 0975 1 025 1 05 1075 1 1 1125 1 15 1175
78. th viability and endpoint and extrapolate the optimal pulse length parameters 3 For multiple pulsing systems protocols vary the number of pulses at the optimal field strength kv cm and pulse length Assay sample for both viability and endpoint Plot the number of pulses versus both viability and endpoint and extrapolate the optimal number of pulses Page 39 www btxonline com ECM 630 Electroporation System Appendix D Electrical Troubleshooting Instrument Does Not Power Up Verify that the power cord is fully inserted in the instrument and in the wall outlet Verify that the fuse is not blown Disconnect power cord from the instrument before removing the fuse holder Replace the fuse if necessary with same rated fuse as indicated on back panel Unanticipated RC time constants or peak voltage output Un anticipated RC time constant or peak output voltage may be a sign that appropriate capacitors are not being selected or are not fully charging Please remember that the external load sample reduces the expected time constant and voltage to various degrees If you believe there is a problem contact BTX Technical Support for immediate consultation LCD Error Messages The 630 is constantly monitoring the parameters of some of its internal circuitry In the case of a malfunction one of the following messages will appear on the display Note the instructions on the following page used to confirm the absence of a pulse
79. ther 25 pF or 50 pF 25 to 1575 with 25 Q resolution Over 200 ECM 600 protocols may be duplicated with this instrument The additional 630 resistor selection will allow researchers to reproduce protocols from competitive systems lacking resistor settings or reporting unlimited resistance The 630 will perform the widest range of electroporation applications among commercially available electroporation generators www btxonline com ECM 630 Electroporation System Quick Start Installation 1 Install on a bench or work table 2 Allow a 1 to 2 inch clearance for proper cooling It is normal for the instrument to be slightly warmer than its operating environment 3 Choose an outlet that is readily accessible Connecting 1 Insert female end of power cord into male power interface on the back panel of the ECM 630 2 Plug male end of power cord into appropriate electrical outlet 3 Insert male banana plugs of the 630 B Safety Stand or alternative electrode device into High Voltage output located on front panel of the 630 Initializing 1 Push the power switch located on the front panel of the 630 2 The 630 will go through a series of self test algorithms to test generator functionality 3 The display will flash 4 Following this initialization screen the first time the instrument is initialized the factory default display will then
80. transfer of materials into and out of the cells During a typical electroporation process target cells and molecules are mixed together When an electroporation pulse is delivered the result is the formation of temporal pores Before the pores reseal the target molecules are observed to enter the cells Upon resealing of the pores the molecules become incorporated within the cell The eventual target site depends on the application for example molecules can remain in the cytoplasm interact with the membrane and move Electroporation Process into the nucleus Applications for electroporation include permeabilization of virtually all cells to a wide variety of molecules and ions The most common applications for electroporation are the transformation or transfection of cells with DNA or RNA Other applications for electroporation include electroactivation electroinsertion of proteins into cell membranes and electroextraction of molecules from cells Although electroporation has mainly been used as a research tool recent work has demonstrated its potential for clinical applications Some areas being explored include electrochemotherapy which involves electroporation for delivering chemotherapeutic agents directly to tumor cells encapsulation of drugs genes into cells for their use as carrier systems transdermal delivery of drugs genes gene therapy and delivery of drugs genes with an electroporation catheter Electr
81. ty EC and Low Voltage Directive 73 23 EEC for Product Safety Compliance was demonstrated to the following specifications as listed in the Official Journal of the European Communities EN 50081 1 Emissions EN 55011 Class B Radiated and Conducted Emissions EN 55082 1 Immunity IEC 10004 2 Electrostatic Discharge Immunity IEC 10004 3 RF Electromagnetic Field Immunity IEC 10004 4 Electrical Fast Transient Burst Immunity Low Voltage Directive 73 23 EEC Page 8 www btxonline com ECM 630 Electroporation System Introduction The 630 is a new state of the art exponential decay wave electroporation system designed for in vitro and in vivo electroporation applications This system incorporates features that make it the most technically advanced exponential decay electroporation system currrently available ECM 630 features include Page 9 The generator utilizes the new BTX Power Platform Technology design and novel digital user interface The revolutionary Precision Pulse System provides the researcher unparalleled power in controlling the time constant With the ability to deliver a maximum of 6000 A in the Low Voltage Mode the 630 is the most powerful generator in its class Low voltage range of 10 V to 500 V with 1 V resolution and 1 pF 25 pF to 3275 pF in 25 pF increments 25 Q to 1575 25 resolution with setting High voltage range of 50 V to 2500 V with 5 V resolution and ei
82. ur bill will include return shipment freight charges Disassembly by the user is prohibited Service should only be carried out by experienced BTX HARVARD APPARATUS technicians Repair Facilities and Parts BTX Harvard Apparatus stocks replacement and repair parts When ordering please describe parts as completely as possible preferably using our part numbers If practical enclose a sample or drawing We offer complete reconditioning service Page 4 www btxonline com ECM 630 Electroporation System General Safety Summary Review the following safety precautions to avoid injury and prevent damage to this product or any products connected to it To avoid potential hazards use this product only as specified Only qualified personnel should perform service procedures To Avoid Fire or Personal Injury USE PROPER POWER CORD Use only the power cord specified for this product and certified for the country of use CONNECT AND DISCONNECT PROPERLY Do not connect or disconnect probes or test leads while they are connected to a power source GROUND THE PRODUCT This product is grounded through the grounding conductor of the power cord To avoid electric shock the grounding conductor must be connected to earth ground Before making connections to the output terminals of the product ensure that the product is properly grounded OBSERVE ALL TERMINAL RATINGS To avoid fire or shock hazard observe all ratings and markings on t
83. uring Warranty 1 Write or call the HARVARD APPARATUS Customer Support Group and describe the nature of the problem 2 Carry out minor adjustments or tests as suggested by HARVARD APPARATUS 3 If proper performance is not obtained BTX HARVARD APPARATUS will notify you to ship the instrument prepaid to its Service Department The instrument will be repaired and returned at no charge for all customers in the continental United States Customers outside of the continental United States who have purchased our equipment from distributors should contact the distributor If you have purchased your equipment from us you should contact us directly We will repair at no charge but will not pay for shipment documentation etc These charges will be billed at cost Note Under no condition should the instrument or accessories be returned without prior approval from BTX HARVARD APPARATUS An RMA Returned Materials Authorization number must be obtained Page 3 www btxonline com ECM 630 Electroporation System General Information continued Out Of Warranty Service Proceed exactly as for Warranty Service above If our Service Department can assist you by phone or correspondence we will be glad to at no charge Repair service will be billed on the basis of labor and materials A complete statement of time spent and materials used will be supplied Shipment to BTX HARVARD APPARATUS should be prepaid Yo
84. ver the electroporation pulse s to the sample Warning Use proper eye protection during electroporation 5 Remove the Dish Electrode carefully clean as appropriate and prepare for next experiment Petri Pulser 1 Plug the HV cables from the Petri Pulser into the voltage output of the BTX Generator Warning Make sure the Generator is turned off prior to connecting any cables to it If using the Enhancer 3000 to monitor output connect the banana plugs into the output ports of the High Voltage Probe Use the black and red high voltage cables to connect the BTX generator to the input ports of the Enhancer 3000 2 Prepare sample for electroporation in 6 well plate or in 35mm Petri Dish Appendix L Connecting Electrodes Continued 3 Place the electrode in one well of the 6 well plate or 35mm Petri Dish Allow it to gently rest on the surface of the dish 4 Following instructions for the BTX generator and set the appropriate parameters 5 Deliver the electroporation pulse s to the sample Warning Use proper eye protection during electroporation 6 Remove the Petri Pulser carefully clean as appropriate and complete delivery of pulses to all wells if using a 6 well plate Page 51 www btxonline com ECM 630 Electroporation System Appendix M Plate Handler Model HT 100 Plate Handler 1 Select a plate size 3 Select 4mm or 2mm gap plate For example if currently using 4 mm cuvettes use a 4 mm plate

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