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1. 4 9375 _ 5625 553125 500 826875 66375 700605 7315 7714375 81125 848125 450 4875 525 562 5 600 6375 675 7125 750 787 5 825 8825 4575 405825 _ 53375 571875 610 648 125 68825 724375 7825 80068 83875 876875 465 50375 5425 56125 520 65875 1975 73625 ms 81375 8525 801 25 4725 511875 _ 55125 _ 590625 630 _ 869875 _ 70875 748125 7875 826875 _ 06625 905825 Page 29 www btxonline com ECM 630 Electroporation System Appendix B ECM 630 RC Time Constants continued Low Voltage Mode RC 1123 75 625 850 675 700 725 750 775 100 B25 450 875 100 15625 16 25 16875 5 a 1875 19375 wes as 5 725 3125 325 3875 35 3525 375 3875 40 41 25 225 4375 a5 46 075 48 75 50625 525 5625 50105 6 1 075 8075 65605 es 625 65 075 m 725 75 775 a 825 85 a5 90 78125 8125 LI 5 975 9875 100 103125 10625 1035 nas 9375 EZ 101 25 105 108 75 n25 ns 120 128 75 1375 13125 135 100 375 11978 15 1225 10687 190125 156 140 144375 14875 153125 1575 125 190 135 140 145 150 156 180 185 m 180 140 625 14625 151 675 1675 165 125 106 75 174 375 180 185 625 18125 196 875 2025 15625 1625 168 75 175 181 25 1875 19575 200 20625 2 25 25 171678 wars 1925 35 20625 5 20 1226875 23375 240685 247 5 1875 106 2025 210 2175 225 2325 240 2475 255 2825 270 203 125 amas 210975 275 085005 24076 251875 280 268185 27695 284375 2905 218 75 2275 23525 245 25375 626 271 280 288 75 as 30625 315 24075 wes 271875 2815 20005
2. 0505 88875 710875 TAD 268125 70025 sass gaps 59375 675 61 665 88875 7125 736 25 760 78375 8075 8315 455 609375 63075 68581125 s 705875 7825 755005 Bas 82875 ASIN ams 825 850 875 700 725 750 ns BOO B25 850 875 900 640 625 86626 601875 775 743125 76875 5 820 845626 87125 806875 9225 650 25 6825 708 75 735 75125 187 5 81375 LJ 866 25 es 918 75 LJ 671875 80875 725625 725 770375 80625 833 125 160 885875 1378 940685 9675 8875 Ll 7425 ma 7075 LI 8528 LJ 9075 95 9825 190 703 125 731 25 85 705 815625 84875 871805 wo 928125 95625 984375 10125 71875 7475 768 805 83375 862 5 5 920 94875 975 10625 1035 724375 708875 780195 25 ames ama 910005 mo 969375 99675 1028125 10575 750 780 Bio 840 o 900 930 360 990 1020 1050 1080 765 625 79625 895875 8575 885 125 1675 949375 wo 100895 10125 1071875 11025 78125 m25 84375 75 006 25 975 6475 1000 109125 10625 109375 1125 796875 40875 REDG25 mos sus 95025 985175 1080 1051875 108375 1015605 11475 8125 Bas ams E 3425 975 10075 1040 10725 1105 11375 1170 828 125 86125 amas 95 060625 00475 1026875 1060 1003125 112625 1158375 nga 4375 875 1 145 97875 10125 100625 1080 1375 5 110125 1215 880 375 80075 gaies 25 00685 103125 106562 noo 1184375 116875 1203126 1287 5 875 mo 145 080 1015 1050 1086 nao 1156 1180 860 625 92626 061875 9975 1035195 106875 1104378 mao 406 25 mas 97805 1005 105125 10675 112375 NGO 801875 96875 85 103
3. User s Manual ECM 630 Electroporation System 6 cwo Cell Manipulator Ele MA1 45 0051 ECM 630 Electroporator only 110V MA1 45 0051int ECM 630 Electroporator only 220V HARVARD APPARATUS The Electroporation Experts Publication 5501 003 REV C 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 9 and a
4. 1082 5 1125 1187 5 1250 1912 5 1375 14375 1500 1562 5 1625 16875 1750 1812 5 1875 19375 20625 2125 21875 23125 2375 2437 5 25605 26875 2750 2812 5 2875 2937 5 3062 5 3125 31875 39125 3375 3437 5 35625 36875 3750 38125 3875 3937 5 2525 63 125 126 25 189 375 2525 315 625 378 75 441 875 505 568 125 531 25 694 375 7575 820 625 883 75 946 875 1010 1073 125 1136 25 1199 375 12625 1325 625 138875 1451 875 1515 1578 125 1641 25 1704375 1767 5 1830 625 1893 75 1956 875 2020 2083 125 2146 25 2209 375 22725 2335 625 2398 75 2481 875 2525 2588 125 2651 25 2714375 27775 2640 625 2903 75 2966 875 3030 3093 125 3156 25 3219375 32825 3345 625 3408 75 3471 875 3535 3598 125 3661 25 5724375 37875 3850 625 391375 3878875 www btxonline com ECM 630 Electroporation System Appendix B ECM 630 RC Time Constants continued Low Voltage GAPAN AP Mode RC 25 00 083 1 25 50 00 1 25 2 50 75 00 188 375 3225 3250 100 00 250 5 00 80 525 81 25 125 00 313 6 25 151 25 1625 150 00 3 75 7 50 241 875 243 75 24562 175 00 4 38 8 75 3225 325 4 200 00 5 00 10 00 403 125 406 25 408 225 00 5 63 11 25 483 75 487 5 250 00 5 25 12 50 a 73 105 275 00 6 88 13 75 ms 7888 4 18 806 25 812 5 gt 885 875 893 75 900 6 25000 875 wee E 2675 975 375 00 9 38 18 75 5 1048125 105625 1064 400 00 Rm am A 1128 7
5. 1125 1170 1215 1260 1305 1350 1385 Map 14B5 1620 1605 1710 1755 1800 1845 1890 1035 1980 2070 2115 2160 2205 2250 2385 2430 2475 2565 2610 2700 2745 2790 2835 8 56 5 8 5 8 8 8 1826 1850 45 625 4625 9125 925 136875 13875 1825 185 228125 23125 27375 277 5 319375 32375 365 370 4065 416 26 45625 4525 801 875 60875 547 5 555 588125 60126 83875 6475 684 375 60375 730 77505 786 25 B2125 e925 886 875 878 75 9125 925 958 125 971 25 100875 1017 5 1049375 1063 75 1005 1110 1140 825 1186 25 1186 25 1202 5 1231875 124875 12775 1295 13231255 1341 25 136875 1387 5 1414375 143375 1460 1480 1505625 1525 25 1551 25 1572 5 15908875 75 16425 1665 1686125 1711 25 173375 17575 1779875 180075 1825 1850 1870805 1898 25 1916 5 19425 1961875 198875 2007 5 2034 2053125 2081 25 208875 21275 2144375 217375 2190 2220 2235005 2263 25 2281 25 22125 2026875 295075 28725 2405 2418 125 245125 246375 2497 5 2509375 254375 2585 2590 2600625 263825 2646 25 26825 2691 875 _ 272875 21315 275 2783125 2821 25 282875 _ 28675 4 975 291375 1875 45875 9375 140 625 187 5 234 375 281 25 328 125 375 421 875 458 75 515 605 5625 609 375 856 25 703 125 750 796 875 84375 890 625 8375 984075 1081 25 1078 125 1125 1171 875 1218 75 1265 625 15125 1359 375 1406 25
6. 1453125 1500 1545875 159375 16401825 1687 5 1724375 1781 25 1828125 1875 1921 875 1088 75 2015625 2062 5 2109 375 2156 25 2208 125 2250 2295875 234375 2390 625 2437 5 2484 375 2531 25 2573 125 2025 2671 875 2718 75 2765 625 2812 5 2950375 2906 25 2953 125 www btxonline com ECM 630 Electroporation System Appendix B ECM 630 RC Time Constants continued Low Voltage Mode RC 1925 1950 1975 2000 2025 2050 2075 2100 2126 2150 2175 2200 2225 48 125 4875 40375 50 50605 51 25 51 875 95 53 195 4275 54375 56 _ 55625 96 25 975 9875 100 10125 1025 10875 105 10625 75 10875 110 111 25 14375 14625 148125 180 151 875 15375 185808 1575 169 375 16125 5 166 _ 166875 1925 196 1975 200 2025 205 2075 210 2125 215 2175 220 2225 240625 24375 246875 250 253 125 256 25 259 375 2625 265 675 26875 271875 z5 278125 288 75 2925 2925 300 30375 3075 311 25 85 31875 325 326 25 380 3375 336875 34125 345025 350 354375 35075 363 125 3075 371875 _ 37625 380025 as 380375 385 390 305 400 405 410 415 420 426 1430 435 440 145 433195 43875 444375 45D 455605 46125 465875 4725 4781235 48375 489375 495 _ 500625 481 25 4875 49175 500 50625 5125 51875 526 59125 575 54375 550 556 25 529375 53625 543125 550 556875 56375 570605 5775 584375 59125 508125 605 611875 E 5775 585 5325 600 607 5 615 6225 630 6375 645 6525 660 6575 9 525695 5
7. 3915 41175 4185 42925 2725 68 125 13625 204 375 2725 340 625 40875 476 875 545 613 125 68125 749 375 8175 885 625 95375 1021 875 1090 1158 125 122625 1294 375 13625 14501625 149875 1566 875 1635 1703 125 177125 1839 375 1907 5 1975 625 204375 2111 875 2180 2248 125 231625 2984975 24525 2520 625 258875 2656 875 2725 2783125 2861 25 2020 375 20915 3085 625 313875 3201 875 3270 936 125 340625 3474375 35425 3610625 367875 3746875 3815 3883 125 3951 25 4019 375 4087 5 4155625 222375 4291 075 2750 0875 1375 26125 2681 25 2818 75 2887 5 2956 25 3093 75 31625 3231 25 3366 75 3437 5 3506 25 3643 75 37425 3781 25 3918 75 3987 5 4056 25 4125 4193 75 4262 5 4331 25 2775 69 375 198 75 208 125 2775 346875 41625 485 625 555 624 375 69375 763 125 8325 901 875 971 28 1040 25 1110 1179 375 1248 75 1318 125 18875 1456 875 1526 25 1595 825 1685 1734 375 1803 75 1873125 19425 2011 875 2081 25 2150 625 2220 2289 375 235875 2428 125 24975 2586 875 2636 25 2705 625 2715 2844 3 5 291375 2983 105 30525 3121 875 318125 3200 625 3530 3309375 346875 3538 125 36075 3676 875 314825 3815 825 3885 3054375 4023 75 4083 125 41625 4231 875 4301 25 4370 625 www btxonline com 2800 TO 140 10 8388888888 2 a 583 1190 2825 70 025 141 25 211 875 2825 353 125 423 75
8. 494 375 565 635 625 706 25 776 875 8475 918 125 985 75 1059 378 1130 1200 828 127125 1941875 14125 1483 125 155375 1624375 1695 1765 825 189625 1908 875 19775 2048 125 2118 75 2180 375 2260 2330 625 2401 25 2471 875 25425 2613 125 268375 2754375 2825 2805 605 2966 25 3036 875 31075 3178 125 324875 3319 375 3390 3531 25 3501 875 35725 3743 125 3813 75 3884 375 3955 4025 625 4096 25 4168875 42375 49081125 4376 75 4449 375 7125 1425 21375 4275 49875 570 64125 7125 783 75 926 25 9975 106875 1140 121125 12825 135375 1425 1496 25 15675 1638 75 1710 178125 18525 192375 206625 21975 2208 75 2351 25 2422 5 249375 2636 25 27075 277875 2921 25 29825 306375 3135 320625 3277 5 3348 75 1491 25 35625 363375 177625 3847 5 391875 406125 41325 420375 4275 434025 44175 440875 2875 71875 14375 215 625 287 5 358 375 431 25 503 125 575 646 875 718 75 790 625 8625 934375 1006 25 1078 125 1150 1221875 1293 75 1365 625 1375 1509 375 1581 25 1653 125 1725 1796 875 1868 75 1940 625 20125 2084 375 2156 25 2228 125 2300 2371875 2443 75 2515 625 2587 5 2659 375 2731 25 2803 125 2875 2946 875 301875 3090 625 31625 3234375 3306 25 3378 125 3450 3521 675 359375 3665 625 3737 5 3808 375 3881 25 3953 125 4025 4096 875 4168 75 4240 625 43125 4384 375 4456 25 4528 126 ECM 630 El
9. 85 1275 170 2125 2975 3825 4675 510 5525 5375 1725 43 126 8525 129 375 1725 21565 25875 301 875 345 388 125 43125 474 375 5175 5601625 603 75 646 875 690 733 125 77825 819 375 8625 905 625 94875 901875 1085 1078 125 112125 1164 375 1207 5 12501625 120375 1336 875 1380 1423 125 1466 25 1509 375 15525 1595625 1838 75 1681 875 1725 1788 125 1811 25 1854 375 1897 5 1940 625 1963 75 2026 875 2070 2113 125 2156 25 2189 375 22425 2285 625 2328 75 2371 875 2415 2458 125 2501 25 2544 375 25815 2630 625 2673 75 2716875 91875 1008 25 109375 1137 5 1181 25 1225 1268 75 13125 1356 25 1400 1443 75 1487 5 1531 25 1575 1618 75 16825 1706 25 1750 1793 75 18375 1881 25 1905 1968 75 20125 205525 2100 214375 21875 2231 25 2275 231875 23525 2406 25 2450 2493 75 2537 5 2581 25 2625 2688 75 27125 2756 25 1775 44 375 8875 133 125 1775 221875 25625 310 625 355 392375 75 443 488 125 5325 578 875 62125 665 625 754375 79875 843125 8875 931 875 97625 805 1020 1065 1102 375 1153 75 1198 125 12425 1285 875 1331 25 1375 625 1420 1464 375 75 1508 125 1553 1597 5 1541 875 1888 26 1730 825 1775 1819 375 185375 1908 125 19525 1995875 2041 25 625 2085 2130 2174375 221875 2263 125 2307 5 2351 875 2396 25 2440625 2485 2529375 257375 2518 125 2825 2705875 2751 25 2795625 1800 ad 1 288 1080
10. Match these as closely to settings allowed with the ECM 630 For low voltage protocols V gt 500V if there is no resistance outlined use the ECM 630 resistance setting none For additional recommendations please contact BTX Technical Support Page 19 www btxonline com ECM 630 Electroporation System Advanced Operation Programming 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 and 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 co
11. 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 To 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 15 www btxonline com ECM 630 Electroporation System Quick Start Continued Capacitor 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 observe
12. 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 10 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 ECM 630 Initializing 1 Page 11 Push the power switch located on the front panel of the ECM 630 The ECM 630 will go through a series of self test algorithms to test generator functionality The display will flash Following this initialization screen the first time the instrument is initialized the factory default display will then read The first page of the Set Parameter Screen from the last time the ECM 630 was used will be displayed each time the ECM 630 is powered up after this initial start up www btxonline com ECM 630 Electroporation S
13. 13 75 21 5 41 25 55 68 75 82 5 96 25 110 123 75 1375 151 25 57500 0 575 14375 2875 43 125 575 71 875 8525 100 525 115 129 975 14375 158 125 60000 05 15 30 45 60 75 90 105 120 135 150 165 62500 0 625 15 625 31 25 46875 625 78 125 93 75 109 375 125 140 625 156 25 171875 650 00 0 65 18 25 5 4875 65 8125 975 11375 190 14625 182 5 17875 67500 0 675 18875 3375 50 625 675 84375 104 25 118 125 135 151 875 168 75 185 625 700 00 07 5 35 525 70 875 105 1225 140 1575 175 1925 72500 0725 18 125 3625 54 375 725 90 625 10875 _ 126875 145 183 125 181 25 1958375 75000 075 1875 375 5825 75 9375 1125 131 25 150 16875 1875 206 25 77500 0775 19375 3875 58 125 775 956875 11625 135625 155 174375 19375 213125 80000 0 8 20 40 60 Bo 100 120 140 160 180 200 220 82500 0 825 20 625 4125 61 875 825 103 125 12375 144375 185 185 625 20825 226 875 85000 0 85 2125 425 63 75 85 106 25 1275 14475 170 19125 2125 23375 87500 0 875 21875 4375 65 625 875 108 375 131 25 153 125 175 196 875 218 75 240 625 0000 0 8 225 45 67 5 BO 1125 135 1575 180 2025 225 247 5 92500 9 925 23 125 46 25 69 375 925 115625 13875 161 875 185 208 125 231 25 254 375 950 00 0 95 2375 475 7125 95 118 75 1425 16825 190 21375 2375 261 25 87500 0975 24375 4875 73 125 975 121 875 145 25 170 625 195 219 975 24375 268 125 1000 00 1 25 50 75 100 125 150 175 200 205 250 275 1025 00 1 025 25 625 5125 76 875 1025 128 125 15375 179 375 205 230 625 256 25 281875 1050 00 1 05 26 25 525 78
14. 2828 75 2874 375 2451 25 2497 5 254375 269625 2682 5 272875 282125 28675 291375 796 875 843 75 890 625 9355 984375 1081 25 25 1125 1171875 1218 75 1265 625 15125 1350 375 1406 25 1453 125 1500 1546875 159375 1840 625 16875 1734375 1781 25 1828 125 1878 1921 875 1968 75 20151825 20805 2105375 2156 25 2200 125 2250 296 875 234375 2390 625 2437 5 2484375 263125 2578 125 2625 2671875 2718 75 2765625 28025 2856 375 2906 25 2953 125 www btxonline com ECM 630 Electroporation System Appendix B ECM 630 RC Time Constants continued Low Voltage Mode RC 04 370 128 75 193 125 2575 321 875 386 25 450 625 515 643 75 708 125 836 875 901 25 965 625 1030 1094 375 115875 1223 125 12875 1351 875 1416 25 1480 625 1545 1609 375 167375 1738 125 1802 5 1865 875 1931 25 1995 625 Resistor ohm 2121375 2188 75 2253 125 23175 2381 875 2448 25 2510 625 2575 2639 375 2703 75 2768 125 28325 2895 875 2901 25 3025 825 54 575 3218 75 3283 125 38475 3411 875 3476 25 3540 625 3669 375 373375 3798 125 38625 2920 875 3891 25 4055 605 Page 36 832 58 8 2625 65 625 131 26 196 875 2825 328 125 36375 459375 525 590 625 656 25 721 875 787 5 853 125 918 75 984 375 1050 1115625 1181 25 1246 875 15125 1378 125 1443 75 150 375 1575 1540 625 1706 25 1771 875 1637 5 1903 125 1968 75 2034 375 2100
15. 2925 316875 34126 365625 390 414375 43875 463125 4875 _ 511875 _ 53625 560625 300 325 350 375 400 425 450 475 500 525 550 575 3075 303125 _ 35875 384375 410 495605 46126 486875 5125 _ 588 125 _ 656975 589375 315 341 25 3875 39375 420 446 25 4725 498 75 525 551 25 5775 80375 3025 349375 37625 403 125 430 456875 4B375 510625 53 5 564375 59125 618125 390 357 5 385 4125 440 467 5 495 5225 550 5775 805 6325 3375 365605 99375 421875 450 _ 478125 50625 _ 534375 5825 590625 61875 646875 345 373 75 4025 431 25 460 488 75 5175 _ 54625 575 60375 6325 68125 3605 381875 41126 440625 470 499375 52875 558 125 5875 616875 64625 675625 360 380 420 450 480 510 540 570 600 830 a60 680 3675 398125 42875 459375 490 _ 520625 _ 55125 581875 6125 643125 _ 67375 704375 375 40625 4375 46875 500 53125 5625 59375 625 656 25 68 5 71875 3825 414375 44525 478125 510 541875 57375 605625 6375 669375 70128 733125 390 4225 455 487 5 520 5525 585 617 8 650 6825 715 7475 3075 430625 46375 405875 530 563 128 59625 629375 6625 _ 695625 72875 761875 405 488 75 4725 _ 50625 540 57375 8075 64125 67 5 70875 7425 _ 77625 4125 446875 48125 _ 515625 550 584375 _ 61875 653125 687 5 721875 _ 75825 790625 420 455 490 525 560 595 630 665 700 735 770 805 4275 463125 49875 534 375 570 605625 641 25 676875 7125 748 125 78375 819375 485 _ 47125 5075 _ 54375 580 61625 6525 _ 68875 786 76125 7975 83375 4425
16. 300 5 250 200 0 01 0 1 1 10 Pulse Length ms 366 8 mL HH 366 15 mL Figure 3 Operating Range for the ECM 630 with 366 in LV with PBS at various volumes Operating Range for PP35 PBS LV Petri Pulser gt 20 to 400V 450 10 usec to 40 msec gt 400 5 Ej 350 300 250 200 0 001 0 01 0 1 1 10 100 Pulse Length ms PP35 2 1mL PBS a PP35 2 3mL PBS Figure 4 Operating Range for the ECM 630 with PP35 in LV with PBS at various volumes Operating Range for 610 Water HV 1mm gap cuvette 50 to 2500 V 680 usec to 36 msec 3000 2800 R 25ohm R 100 ohm 2600 C 25uF C 50uF 2400 2200 2000 1800 Voltage V 1600 1400 1200 1000 0 68 5 36 Pulse Length ms E1610 50 uL Figure 5 Operating Range for Model 610 H 0 HV Page 25 www btxonline com ECM 630 Electroporation System Appendix A ECM 630 Electrode Operation Ranges continued Operating Range for 620 Water HV 2mm gap cuvette 2750 50 to 2500 V 2625 R2 25 ohm R 100 ohm R 1575 ohm 660 usec to 38 msec 2500 2375 S 2250 2125 2000 0 66 5 38 Pulse Length ms 620 200 uL Figure 6 Operating Range for Model 620 H 0 HV Page 26 www btxonline com ECM 630 Electroporation System Appendix B ECM 630 RC Time Constants SEE PAGES 28 38 Page 27 www btxonline com ECM 630 Electroporation System Appendix B ECM 630 RC Time Constants
17. 375 1710 1745 525 1781 25 1815875 18525 1888 125 1923 75 1959 375 1995 2090 625 2066 25 2101 875 21375 2173125 920875 2244 975 1430 3625 725 10875 145 18125 2175 25375 97875 1015 105125 1067 5 112375 1160 119525 12325 126875 1305 1341 25 1377 5 141375 1450 1488 25 15225 1558 75 1595 163125 1667 5 170375 1740 1776 25 16125 184875 1685 1921 25 19575 199375 2030 206625 21025 2138 75 2175 221125 22475 228375 1475 35 875 7375 110625 1475 184375 221 25 258 125 205 331 875 368 75 405 625 442 5 479 375 516 25 553 125 590 526 875 663 75 700 625 7375 774 375 81125 848 125 885 921 875 958 75 995 625 10825 1069 375 1106 25 114 125 1180 1216 875 125375 1280 625 18275 1364 375 1401 25 1438 125 1475 1511875 1548 75 1585 025 10225 1659 375 1696 25 1733125 1770 1806 875 1843 75 1880 625 19175 1954 375 1981 25 202 125 2065 2101 875 2138 75 2175 625 22125 2249 375 2286 25 2929125 www btxonline com 1500 75 75 112 5 187 5 262 5 337 5 375 4125 4875 525 562 5 0375 675 7125 750 787 5 825 862 5 900 937 5 975 1012 5 1087 5 1125 11625 12375 1275 13125 1350 1387 5 1425 14625 15375 1575 16125 16875 1725 17625 1800 1837 5 1875 1912 5 1950 1987 5 20625 2100 21375 215 22125 2287 5 23625 1525 38125 78 25 114 375 152 5 190 625 228 75 266 875 305 343 125 381 25 419 375 457 5 4
18. 75 105 131 25 1575 183 75 210 236 25 262 5 28875 1075 00 1 075 26 875 5375 80 625 1075 134375 18125 __ 188 125 25 241 875 26875 _ 285025 1100 00 11 27 5 55 825 110 1975 165 1925 220 2475 275 wes 1125 00 1125 28 125 56 25 84 375 1125 140 625 168 75 196 875 225 253 125 281 25 309 375 1150 00 1 15 2875 55 86 25 115 143 75 1725 201 25 230 258 75 287 5 31625 1175 00 1475 29375 5875 88 125 175 148875 17625 205625 285 264 375 29375 323125 1200 00 12 30 60 90 120 150 180 210 240 270 300 330 1225 00 1225 30 625 61 25 91 875 1225 153 125 18375 214375 245 275 625 306 25 336 875 1250 00 125 31 25 625 9375 125 156 25 1875 2175 250 281 25 3125 343 75 1275 00 1275 31875 6375 95 625 1275 159 375 151 25 223 125 255 286 875 31875 350 625 1300 00 13 325 65 975 130 1625 195 2275 200 925 325 3575 1325 00 1 325 33125 66 25 95 375 1325 105 625 19875 _ 231875 205 _ 298125 33125 2304375 1350 00 135 3375 5 10125 135 188 75 202 5 235 25 270 30375 3375 371 25 1375 00 1375 34375 6875 103 125 1375 171 875 206 25 240 625 275 309 375 343 75 376 125 1400 00 14 35 70 105 140 175 210 245 280 35 350 385 1425 00 1425 35605 7125 106 875 1425 178 125 213 75 249375 285 320 625 356 25 391 875 1450 00 145 3525 725 10875 145 181 25 275 25375 280 326 25 3625 39875 147500 1475 36875 7375 110895 1475 184 375 221 25 258 125 285 331 875 36875 405 625 1500 00 15 375 75 1125 150 187 5 225 2025 300 2375 375 4125 1525 00 1525 38 125 76 25 114375 1525 190 625 228
19. 75 266 875 305 343 125 38125 419 375 1550 00 155 3875 75 11625 155 19375 2325 27125 310 348 75 3875 426 25 1575 00 1 575 39 375 7875 118 125 1575 190 875 230 25 275 625 315 354 975 39275 433125 Page 28 www btxonline com ECM 630 Electroporation System 300 325 350 375 400 425 450 475 500 525 560 575 75 8 125 875 9375 10 10625 1125 11875 125 13125 1375 14375 15 1625 175 1825 20 2125 225 2375 25 2625 215 28 75 225 24375 2625 28125 3 3185 3378 _ 35625 3 5 30375 125 43125 30 325 35 35 40 425 45 475 so 525 55 575 975 40625 4375 46875 S0 53125 5625 _ 59375 625 65 625 8875 71875 45 48 75 525 56 25 50 6375 675 7125 75 78 75 825 86 25 525 56875 6125 65625 70 74375 7875 83128 ars 91875 9625 100625 60 65 70 75 B0 85 90 95 100 105 n0 n5 625 73125 7875 84375 90 95625 10125 108875 1125 118 125 12375 129375 75 B125 875 9375 100 10625 1125 11875 125 19125 137 5 14375 825 89 375 9625 108125 10 118875 12376 130625 1325 144375 15125 158125 90 975 105 1125 120 1275 135 1425 150 1575 165 1725 975 105625 11375 _ 121875 130 138125 14825 154375 1825 170625 17875 186875 105 11375 1225 13125 140 14875 1575 16525 1775 18375 1925 20125 1125 121875 13125 140625 150 150375 16875 178125 1875 195875 20625 215625 120 120 140 150 160 170 180 190 200 210 220 230 1275 190125 14875 159375 170 180625 19125 201875 2125 228125 29975 244375 105 14625 1575 1687
20. Glossary of Electrical Terms Appendix G Glossary of Biological amp Technical Terms 43 44 Appendix H Electroporation Generator Compatibility reeennn Appendix I Recommended Reading Appendix J Accessories and Replacement Parts 47 Appendix K General Care and Cleaning 48 Page 1 www btxonline com ECM 630 Electroporation System General Information Serial Number The serial number for the ECM 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 BTX Harvard Apparatus warranties the ECM 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 to misuse neglect acci dent 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 fur nished by BTX HARVARD APPARATUS This warranty extends only to the original customer purchaser IN NO EVENT SHALL BTX HARVARD APPARATUS BE LIABLE FOR INCIDENTAL OR CONS
21. 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 BTX HARVARD APPARATUS Holliston Massachusetts facilities or an authorized service site Owner will ship instrument prepaid to Holliston Massachusetts USA or the service site BTX HARVARD APPARATUS will return the instrument after servicing freight prepaid to owner s address Obtaining Service Service During Warranty 1 Write or call the BTX HARVARD APPARATUS Customer Support Group and describe the nature of the problem 2 Carry out minor adjustments or tests as suggested by BTX 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
22. continued Low Voltage Mode RC 1 25 5 75 100 125 150 175 200 225 250 275 2500 0025 0625 125 1875 25 3 125 375 4375 5 5825 625 6875 50 00 0 05 1 25 25 375 5 625 75 875 10 11 25 125 1375 7500 0 075 1875 275 5 625 75 4375 11 25 13125 15 16875 1875 20 625 10000 0 1 25 5 7 5 10 125 15 75 20 225 25 27 5 12500 0 125 3125 6 25 9 375 125 15 625 18 75 21 875 25 28 125 31 25 34375 15000 0 15 375 75 1125 15 1875 225 26 25 30 2375 375 41 25 17500 0175 4375 875 13 125 175 21 875 26 25 30 525 35 39 375 43 75 48125 20000 02 5 10 15 0 25 30 35 40 45 50 55 22500 0 225 5625 1125 16 875 225 28 125 33 75 39 375 45 50 625 96 25 61875 25000 0 25 8 25 125 18 75 25 31 25 37 5 4375 50 5625 625 6875 27500 0 275 6 875 1275 20 625 275 34375 41 25 48125 55 51 875 88 75 75 825 E 30000 0 3 5 15 225 30 37 5 45 525 60 875 75 825 5 32500 0 325 8 125 1625 24 375 325 40 625 48 75 56 875 65 73 125 81 25 88 375 5 35000 0 35 8 75 17 5 2625 35 4375 525 61 25 70 78 75 875 9625 37500 9 375 9375 1875 28 125 375 48 875 5525 65 525 75 BA 375 93 75 103 125 2 40000 0 4 10 20 30 50 60 70 BO so 100 110 0 42500 0 425 10625 21 25 31875 425 53 125 63 75 74 375 85 95 625 106 25 116 875 c 45000 0 45 11 25 25 3375 45 56 25 67 5 78 75 90 101 25 1125 12375 47500 OATS 11 875 2375 35 525 475 58 375 7125 83125 95 106 875 118 73 130625 50000 05 125 25 37 5 50 625 75 875 100 1125 125 137 5 52500 0 525 13 125 26 25 39 375 525 65 625 78 75 91 875 105 118 125 131 25 144 975 550 00 0 55
23. 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 III Products in this Category Distribution level mains USE ON PATIENTS fixed installation CAT II 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 Compatibility 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 ECM 630 is a new state of the art exponential decay wave elec troporation 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 P
24. is again strongly recommended Molecules Introduced by Electroporation DNA RNA dNTPS Enzymes Antibodies Other Proteins Peptides Dyes Sugars lons Other Molecules Page 22 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 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 ECM 630 Electrode Operation Ranges Figures 1 6 display expected operating ranges for the ECM 630
25. 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 to three available programs there is also the default program as outlined in the Initializing section 2 Rotate the knob to change the program number 3
26. 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 Your 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
27. 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 2 mm gap cuvette 575 R 1575 ohm 20 to 500V 800 N3750 ohm sad 25 usec to 93 msec E ag STF 350 X R 25 ohm 275 C 3275 uF 0 01 0 1 1 10 100 Pulse Length ms 620 40 uL PBS 620 400uL PBS Figure 1 Operating Range for the ECM 630 with Model 620 in LV with PBS at various volumes Operating Range for 640 PBS LV 4 mm gap cuvette 600 R 500 ohm R 1575 ohm 20 to 500V C 1 uF C 25 uF 500 a R 25 ohm 25 usec to 70 msec C 3275 uF 400 Voltage V 300 200 0 01 0 1 1 10 100 Pulse Length ms e 640 200 ul m 640 400 uL 34 640 800 uL Figure 2 Operating Range for the ECM 630 with Model 640 in LV with PBS at various volumes Page 24 www btxonline com ECM 630 Electroporation System Appendix A ECM 630 Electrode Operation Ranges continued Operating Range for 366 PBS LV Petri dish electrode 400 R 1575 ohm 20 to 350V usec to 6 msec 25
28. 0 89375 9075 920 924375 94375 960 975 990 1000 1015 625 1031 25 1040 105625 10725 1080 1096875 111375 1120 11375 1155 1160 1178125 119525 1200 1218 75 1237 5 1240 1259375 127875 1280 1300 1320 1320 1340625 1361 25 1960 138125 14025 1400 1421875 144375 1440 14625 1485 1480 160855 182825 1528 154375 18875 1500 1584375 160875 1600 1825 1650 1640 1685625 1691 25 1680 170625 1732 5 1720 1746875 177375 17825 1815 1800 182825 185625 1840 186875 _ 18975 1880 1909375 _ 193475 1990 1950 1980 1980 1990685 2021 25 2000 203125 20825 2040 2071875 210375 200 21125 2145 2120 2153125 218825 2160 219375 22275 2200 2234375 226475 2240 2279 2310 2080 2315625 235125 2320 235825 23925 2360 2396875 243375 240 24375 2475 2440 2478125 _ 2518 25 P480 _ 251875 _ 25575 2520 2859975 259875 Page 33 1675 41 875 83 75 125625 167 5 208 375 25125 293 125 335 378 875 41875 460 625 502 5 544 375 586 25 628 125 870 711 875 75375 796 625 8375 879 375 321 25 963 125 1005 1046 675 1088 75 1130 625 1725 1214375 1256 25 1298 125 1340 1381 675 142375 14 5 625 1507 5 1549 375 1591 25 1639 125 1675 1716875 1758 75 1800 625 18425 1884375 1826 25 1968 125 2010 2051875 209375 2135825 21775 2219 375 2261 25 2308 125 2345 2386 875 2428 75 2470 625 25125 2554375 2586 25 2538 125 1700 425
29. 0 to give a result in msec Page 18 630B MA1 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 ECM 630 resistance setting 4 Set the capacitance from the ECM 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 The ECM 630 is capable of reproducing most exponential competi tive system protocols BioRad Gene Pulser and Gene Pulser II Protocols For protocols using the Capacitance Extender Capacitance Extender Il 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 by 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
30. 050 2278125 230625 2334375 29625 2300625 241875 2445875 2475 2503125 2213 75 2242 5 2271 25 2300 2328 75 23575 2386 25 2415 2443 75 24725 2501 25 2530 2558 75 261 875 2291 25 2320625 9350 2379375 240875 2438125 2467 5 2496875 252625 25556025 2585 2614375 2310 2340 2370 2400 2430 2460 2490 2520 2550 2580 2610 2640 2070 2058 125 _ 298875 2419075 2450 2480 625 251125 2541075 25725 200125 283375 2604375 2005 2725625 240825 24975 245875 2500 253 25 25625 259375 2625 265625 26875 271875 2750 278125 2454375 248625 2518125 2550 2581 875 261375 2645625 2677 5 2709375 274125 2773125 2805 5 25025 9555 _ 25675 2600 _ 26325 2865 26975 2730 27625 2795 28275 260 28925 2550 625 258375 2616875 2650 2683125 271625 2749375 27825 2815625 284875 2881875 2915 2948125 250875 26325 266825 2700 273375 21615 280125 2836 286875 20085 203625 2900 300375 2546875 268125 2715625 2150 2784375 281875 2853125 20875 2921 875 295825 2990625 3025 059 35 2695 2730 2765 2800 2835 2870 2005 2940 2975 9010 3045 3080 3115 2743125 277875 8 2850 2885625 292125 2956875 20025 3028125 306875 3090375 3185 3170625 2791 28 28275 2863 75 2900 2036 25 29725 3008 75 3045 3081 25 3117 5 3153 75 3190 322625 2808375 287625 5 2050 2086875 302375 3060625 30075 3134375 317125 3208 125 3245 3281875 2887 5 2925 29025 3000 30375 3075 31125 3150 31875 3225 32625 3300 3997 5 2935 625 297375 3091875 3060 3088125 312625 3164
31. 10 2280 2315825 235125 2300 28625 23025 2380 2306875 243375 240 2495 2475 2440 2470125 2516 25 2480 251875 25575 2500 2559375 250875 1675 41 875 125 625 1675 209 375 25125 293 125 335 376 875 41875 400 025 5005 544 375 58625 628 105 670 711875 75375 795 625 8375 879 375 921 25 063 125 1005 1046 875 1088 75 130 625 11725 1214 375 1256 25 1298 125 1340 1381875 1423 75 1465 625 15075 1549 375 1591 25 1633 125 1675 1716 875 1758 75 1800 625 18495 1884 375 1926 25 1968 125 2010 2051 875 2093 75 2135625 21775 2219375 2261 25 2303 125 2345 2385 875 2428 75 2470 625 25125 2554375 259625 2638 125 1700 425 127 5 2125 297 5 382 5 4075 510 5525 637 5 722 5 807 5 892 5 9775 1020 1062 5 105 1475 1190 1230 5 1275 1317 5 1360 1402 5 1445 1487 5 1530 15725 1615 1657 5 1700 1742 5 1785 1827 5 1870 1912 5 1955 1997 5 2082 5 2125 2167 5 2210 2252 5 23375 24225 2465 25075 2550 2590 5 29 775 1725 43 125 8625 129 375 1725 215625 258 75 301 875 345 383 125 431 25 474375 517 5 560 625 603 75 646 875 620 733125 77625 819 375 8525 905 625 94875 901875 1035 1078 125 1121 25 1164375 12075 1250 625 129375 1335 875 1380 1423 25 1466 25 150 375 15525 1595625 1638 75 1681875 1725 1768 125 1811 25 1851375 1897 5 1940 625 1983 75 2025875 2070 21 125 2156 25 210 375 2242
32. 2165 625 2231 25 2296 875 23625 2428 125 2498 75 2559375 2625 2690 625 2756 25 2821 875 2887 5 2053 125 3018 75 3084 375 150 3215 625 3281 25 3346 875 34125 3478 125 3543 75 3609375 3675 3740 B25 3806 25 3871875 39375 4003 125 4068 75 4134 375 2650 6625 1825 19875 33125 397 5 453 75 595 25 6625 128 75 861 25 927 5 99375 1060 1196 25 11925 125875 1325 1891 25 152375 1590 1656 25 17225 1788 75 1855 1021 25 19875 2053 75 2120 2186 25 22525 231875 245125 25175 2583 75 2650 2716 25 27825 2848 75 2815 298125 30475 311375 3180 3246 25 33125 337875 3445 351125 35775 3643 75 3710 3776 25 38425 3908 75 3975 404125 41075 417375 2875 66 875 13375 200525 2675 334375 40125 468 125 535 601 875 668 75 735625 8025 889 375 936 25 1003125 1070 1136875 1209 75 1270525 1337 5 1404 375 1471 25 1538 125 1605 1671875 1738 75 1805 525 18725 1930375 2006 25 2073125 2140 2206875 2273 75 2340 625 2407 5 2474375 2541 25 2608 125 2675 2741875 2808 75 2875625 29425 3009375 307625 3143 125 3210 3276575 334375 3410525 34775 3544375 3611 25 3578 125 3745 3811875 3878 75 345 825 40125 4079 375 4146 25 4213 125 2700 675 15625 1687 5 1755 18225 19575 20025 2160 22275 23025 24975 2585 28325 2700 27675 2002 5 30975 3105 31725 3240 3075 3375 3425 3510 35775 37125 3780 3847 5
33. 25 1060375 110625 1149125 1180 9375 975 10125 1050 10875 1925 11025 1200 953 125 99125 1029375 10675 1105628 114875 1801875 1220 86675 10075 104625 1240 1063 Page 30 www btxonline com ECM 630 Electroporation System TREE PERS EN Be 5 1 3 SUSZE 1 LUNA U 870885853 8888993293888888882868888888B23288388883358888888P3E888 5 TUGTOG 8 2 BRSHPRPERER HAG R TIS BAUAN STP ECES ESTEE Ma Lettie ttt ir 1 iti U IIBDPEBEBPBRPBPBHEDPEPEPHPBEIPHBIPIEIHH Miri SES E TERESE STER SE 1 itis AHAHA GEO AG GA HEER TL hail AAAH pnt np Bit BRA UAB 526542545569 F4E5 7 lt www btxonline com Page 31 ECM 630 Electroporation System Appendix B ECM 630 RC Time Constants continued Low Voltage Mode RC 3225 31 875 63 75 95625 1275 150 375 191 25 228 125 265 286 875 31875 350 625 3825 414 375 44625 478 125 510 541 875 57375 605 625 8375 880 375 701 25 783125 765 796875 828 75 860 625 8025 924375 956 25 988 125 1020 1051 875 1083 75 1115625 11475 1178375 1211 25 1243 125 1275 1306 875 1238 75 1370625 14025 1434 375 1406 25 1498 125 1530 1561 875 159075 1625 625 16575 1689 375 1721 25 1753 125 1785 1816 875 1848 75 1880 625 19125 1944 375 1976 25 2008 125 Resistor ohm Page 32 1300 325 87 5 130 162 5 195 227 5 292 5 357 5 402 5 9275 960 625 99375 1020 875 1060 1083 125 1126 2
34. 25 155875 1576875 1505 1613125 144375 14625 148025 1800 151875 15975 155625 1575 150875 16125 163125 1650 166875 1491 875 1511 25 1530625 1850 1569375 188875 5 16275 1646875 166625 1685625 1706 1724375 1540 1580 1580 1600 1620 1640 1650 1680 1700 1720 1740 1760 1780 1588125 100875 1629375 1650 1070825 1169125 1711875 17325 17583125 177375 1794375 1815 1835625 1636 25 16515 167875 1700 172125 17425 176375 1785 1806 25 18275 184875 1870 189125 1684375 1706 25 1728125 1750 1774 875 179375 1015625 18375 1859 375 188125 1903125 1805 1940875 17325 YBS 47775 1800 18225 1845 18675 1890 19125 Mas 19575 1380 20025 1780625 190375 1826875 1850 1873125 189625 1919375 19425 1965625 198875 5 2095 2058125 1828 75 18525 _ 187625 1900 192375 19475 197125 1995 20875 20425 2066 25 290 210375 1876875 _ 1900125 1925625 1850 1974375 199875 2023125 20475 2071875 209625 2120625 2145 2169375 1925 1950 1975 2000 2025 2050 2075 2100 2125 2150 2175 2200 2225 1973125 _ 199875 2024375 205 2075625 210125 2126875 21525 2178125 220375 2229375 2255 2280625 2021 25 20475 207375 2100 212625 20525 217875 2205 209125 22575 228375 2510 233025 2060375 209625 2123125 2150 2176875 220375 2230625 22675 2284375 231125 2938125 2365 2301878 21125 2145 21725 2200 22275 2255 22825 2310 23375 2365 23025 2420 2447 5 2165625 219375 2221875 2
35. 3375 641875 650 658125 8625 674 375 6825 690525 69875 706875 75 723128 673 75 0825 09125 70 T0875 775 72625 735 74315 782 5 76125 770 778 75 2 721875 73125 740625 780 780376 76875 778 125 787 5 796875 80625 815625 825 834375 5 o 780 790 800 810 820 830 840 850 880 870 80 880 818 125 82875 839 375 850 860 625 87125 881 875 8025 NG 125 91375 924 375 935 945 625 c 868 25 8775 888 75 900 911 25 9225 93375 945 95625 987 5 97875 990 1001 25 914375 92525 93B125 950 961875 97375 985625 997 5 1009375 102125 5 1045 1056875 962 5 975 9875 1000 10125 1025 1037 5 1050 10625 1075 10875 1100 1125 1010625 102275 1036875 1050 1063125 107025 1089375 11005 1115625 110875 1141875 1155 008125 1058 75 _ 10725 108625 1100 111275 1278 114125 1155 116875 11825 119625 1210 122375 1106875 112125 1135625 1150 1164375 117875 1198125 12075 1221875 123625 5 1265 1279375 1155 1170 1185 1200 1215 1290 1245 1260 1275 1290 1305 1320 1335 1200425 _ 121875 1294375 1250 1265625 1281 25 1296875 13125 1928125 134375 5 1375 1390625 1251 25 1267 5 128375 1900 131625 13325 194875 1365 138125 19975 141375 1480 144525 1290375 131625 1333128 1350 1366875 138375 1400605 14175 1434375 145125 5 1485 1501875 19475 1365 13825 1400 14175 1435 14525 1470 14875 1505 15225 1540 1597 5 1395625 14375 1431875 1450 1468125 148625 1504375 15225 15406
36. 375 32025 3240605 327875 3316875 3355 3393125 290375 _ 30925 3061 25 3100 313875 31775 321625 3256 399375 33395 337125 3410 344875 3001875 307125 3110895 9150 31008375 _ 300075 3268125 32075 3346875 138025 3495025 3465 3604 375 Page 34 www btxonline com ECM 630 Electroporation System Capacitor uF 1600 1625 1650 aD 40825 4125 80 8 25 825 120 121875 12378 180 1625 165 200 203125 20625 240 24575 2475 280 284375 288 75 320 325 330 380 _ 385625 97125 400 406 25 4125 4a0 440876 45376 480 4875 495 500 528125 53625 560 568 75 5775 600 60835 61875 640 550 860 680 690 525 701 25 720 73125 7425 760 771875 78375 800 B125 825 800 853 126 886 25 880 893 75 9075 020 034 375 048 75 960 975 990 1000 1015825 1031 25 1040 105625 10725 1080 1096875 111375 1120 11375 1155 1160 78125 119625 1200 121875 12375 1240 1258375 127875 1280 1300 1320 1320 19408255 1361 25 1360 138125 14025 1400 1421875 144375 1440 14625 1485 1480 1503125 152825 1520 154375 1567 5 1580 1584375 160875 1800 1825 1650 1840 1665625 169125 1680 170625 17325 1720 1746875 177375 1760 1767 5 1815 1800 1828125 1855 25 1840 _ 186875 1897 5 1880 1909375 193875 1920 1950 1980 1960 1990825 2021 25 2000 203125 20625 2040 2071875 210375 2000 21125 2145 2120 2153125 5 2160 219375 22275 2200 2234375 208875 2240 2275 23
37. 4376 446875 4503125 1375 00 3438 68 75 4515 4550 8 1400 00 35 00 70 00 4595 525 463125 4656 87 1425 00 35 7125 4676 25 47125 4748 1450 00 36 25 72 50 4758875 479375 5 1475 00 3688 73 75 4837 5 4875 1500 00 3750 75 00 4918 125 495625 494 37 1525 00 38 13 76 25 4098 75 50375 507825 1550 00 3875 77 50 5079375 511875 51581 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 manipulation ECM lies in selecting appro priate 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 s
38. 5 1159375 11925 1225 625 1258 75 1201 875 1325 1358 125 1391 25 1424315 14575 1490 625 1523 75 1556 875 1580 1623 125 1656 25 1589 375 7225 1755 625 1788 75 1821 875 1655 1888 125 1921 25 1954 375 1987 5 2020 625 2053 75 2085 875 675 708 75 7425 776 25 810 84375 877 5 925 945 97875 10125 1046 25 1080 111375 475 1181 25 1215 1246 75 12625 181625 1350 138375 14175 1451 25 1485 151875 15525 1586 25 1620 165875 1667 5 172125 1755 1788 75 18225 1856 25 1890 1928 75 1957 5 1901 25 2056 75 2082 5 2126 25 1375 34 375 68 75 103 125 1375 171875 206 25 240 625 275 300 375 343 75 378 126 4125 446 875 48125 515 625 584 375 816 75 653 125 721 875 756 25 790 625 850 375 893 75 928 125 9625 996 875 1031 25 1065 625 1100 1134375 1168 75 1203 125 12375 1271 875 1306 25 1340 625 1375 1400 375 144375 1478 125 15125 1545 875 1581 25 1615 6235 1850 1684 375 1718 75 1753125 1821 875 1856 25 1890 625 1925 1959 375 190375 2028 125 20625 2096 875 2131 25 2165 635 1425 35625 1125 105 875 178 125 21375 249 375 320 625 356 25 391 875 4275 463 125 49075 534 375 5m 605 625 641 25 678 875 7125 748 125 783 75 819 375 855 890 625 926 25 961 875 9975 1033 125 1058 75 1104 375 1140 1175625 120125 1245 875 12825 1318 125 1353 75 1389 375 1425 1460 625 1496 25 1531 875 15675 1603 125 1638 75 1674
39. 5 2285625 2328 75 2371 875 2415 2453125 2501 25 2544375 25875 2630 65 267375 2716 875 48125 568 75 6125 74375 7875 83125 875 918 75 9625 1006 25 1050 1093 75 1137 5 1181 25 1225 13125 1356 25 1400 1443 75 14875 1531 25 1575 161875 16825 1706 25 1783 75 1837 5 1881 25 1925 1968 75 20125 2056 25 2100 2149 75 21875 223125 2318 75 2362 5 2406 25 2493 75 2537 5 2581 25 2668 75 27125 275625 1775 44 375 8875 133 125 177 5 221 875 25 266 310 625 355 399375 44375 488 125 5325 576 875 62125 665 625 754375 79875 843 125 887 5 931 875 97625 1020 625 1065 1109 375 115375 1198 125 12425 1286 875 133125 1375 625 1420 1464 375 150875 1553125 1597 5 1641 75 25 1686 1730 625 1775 1819 375 186375 1905 125 19525 1996 875 2041 25 2085 525 2130 2174375 221875 2263 125 2307 5 2951875 2306 25 2440 625 2485 2529 375 2573 75 2618 125 26625 2708 875 2751 25 2795 625 8858388 a a 7 383 1710 1755 1845 1800 1835 2070 2115 888888 775625 821 25 266 876 9125 958 125 100375 1040 375 1095 1140 525 1186 25 1231 875 1277 5 1323 125 136875 1414375 1460 1505625 1551 25 1506 875 16425 1688 125 173375 179375 1825 1870 625 1016 25 1951 875 2007 5 2053125 209875 2144375 2190 2208 625 2281 25 2326 875 23725 2418 125 2463 76 2509375 2555 2600 625 2648 25 2691 875 27375 2788 125
40. 5 909375 970 1030 625 1091 25 1151 875 12125 1273125 133375 1394375 1455 1515 625 1576 25 1638 875 16875 1758125 1818 75 1879 375 1940 2000 625 2061 25 2121475 21625 2243125 230375 2354 375 2425 2485 825 2546 25 2808 475 2667 5 2728 125 2786 75 2849 375 2910 2970 625 3031 25 3091 875 31525 3213 125 327375 3334 375 3395 3455625 351625 3576875 35375 3690 125 375875 3819375 2450 61 25 1225 18375 3675 428 75 551 25 6125 673 75 8575 91875 104125 11025 116375 1225 128625 13475 1408 75 1470 1531 25 15925 1853 75 1715 177625 1837 5 1898 75 1960 200125 20825 214375 2266 25 23275 2888 75 2511 25 25725 2633 75 2756 25 28175 2878 75 300125 30625 312375 3185 324625 3307 5 3368 75 340125 38525 361375 3796 25 37975 3858 75 2475 61 875 123 75 185 625 2475 309 375 371 25 493 125 495 556 875 61875 680 625 7425 804 375 866 25 928 125 990 1051 875 111375 1175 625 1237 5 1299 375 1361 25 1423125 1485 1546 875 1608 75 1670 625 1732 5 1794 375 1856 25 1918 125 1980 2041 875 2103 75 2165625 22275 2289 375 235125 2413 125 2475 2536 875 2598 78 2680 625 27225 2784 375 284625 2908125 2870 3081 875 309375 3155 625 32175 3279 375 3341 25 3403 125 3465 3526 875 3588 75 3650 825 37125 374 375 383625 3898 125 625 125 187 5 250 3125 375 4375 5625 687 5 750 B125 a75 9375 1000
41. 5 11375 114525 425 00 10 63 21 25 2 1200 375 _ 123875 12281 450 00 11 25 22 50 2 1290 1300 475 00 11 88 23 75 vw 1370825 138125 1991 87 500 00 1250 25 00 a 1451 25 1402 5 1473 525 00 1313 25 25 1531875 _ 154375 155562 550 00 1375 27 50 16125 1625 575 00 14 38 28 75 1893 125 1706 25 1719 37 600 00 15 00 30 00 wie hm aa 2 82599 sa Sues id 650 00 1625 32 50 2015 525 2031 25 2046 87 875 00 1688 39 78 2086 25 2112 5 2125 7 700 00 17 50 35 00 2176875 _ 219375 2210 5 725 00 18 19 2257 5 2275 5 lt 750 00 18 75 37 50 2338125 295825 2374375 gt 775 00 19 38 38 75 241875 _ 24375 245628 5 800 00 20 00 40 00 2499375 25875 2538 125 D 825 00 20 41 25 2580 2600 E 850 00 2125 42 50 2660 525 268125 2701 87 875 00 2188 43 75 2741 25 2782 5 2783 900 00 2250 45 00 2BZLUTS 284375 2085 00 825 00 23 13 45 25 ZEE Z weed 950 00 23 75 47 50 SES Si ane 975 00 24 38 48 75 3144375 _ 316875 _ 3193125 190000 2500 5000 pe 1025 00 2563 51 25 2308625 392125 JASAT 1050 00 2825 52 50 3386 25 34125 9425 1075 00 26 88 53 75 2468875 _ 340375 3520 1100 00 2750 55 00 35475 3575 i 1125 00 28 13 56 25 3628 125 365625 3684 97 1150 00 28 75 57 50 3708 75 37375 376625 1175 00 29 38 58 75 3789 375 3818 75 3848 1 1200 00 30 00 60 00 3870 3900 1225 00 30 63 61 25 39508525 _ 3081 25 4011 87 1250 00 31 25 6250 aoe s k KA 1275 00 31 88 63 75 AES ps 1300 00 3250 65 00 Bits GW id 1825 00 33 13 68 25 az PS 1350 00 33 75 67 50 45
42. 5 180 19125 2025 21375 225 29625 24 5 25875 1425 154375 16825 178125 190 201875 21375 225625 2375 200875 26125 273125 150 1625 175 1875 200 2125 225 2375 250 2625 275 287 5 1575 170625 18375 196875 210 223125 23625 240 375 2625 275625 28875 85 166 17875 1925 _ 20625 220 29375 2475 _ 20125 275 28875 3005 91625 1725 186875 20125 215605 230 244375 25875 273125 2875 301875 31825 330825 180 195 210 225 240 255 270 285 300 315 330 345 1875 203125 21875 250 085695 28125 205875 3125 328 125 34375 359375 195 211 25 2275 24375 200 27625 2925 30875 34125 3875 37375 2025 2190375 23625 252125 270 985875 30375 320825 3375 354975 _ 37125 388 125 210 2275 245 262 5 280 2975 315 3325 350 3675 385 4025 2175 235625 25375 271875 290 308125 32525 344375 3825 380625 39875 416875 225 _ 24375 2625 _ 28125 30 31875 3375 _ 35625 37 39375 4125 43125 2325 251875 2125 290525 310 829875 34875 368125 3875 _ 406875 _ 42625 44525 240 260 280 300 320 340 360 380 400 420 440 460 2475 268 125 28875 308 375 330 350605 O 37125 391875 4125 433125 45375 474375 255 276 25 297 5 318 75 340 361 25 3825 40375 425 446 25 4675 483 75 2625 284375 30625 328125 350 371875 39475 415625 4375 459375 48125 503125 270 2925 315 3375 360 3825 405 4275 450 4725 495 5175 2775 300625 32375 348875 370 303125 41625 40375 4625 485625 _ 50875 531875 285 30875 3325 35625 380 40375 4275 45125 475 49875 5225 54625
43. 8 125 197625 2034 375 20025 2150625 2208 15 2266 875 2325 2383 125 244125 2499 375 25575 2515 625 267375 2731 8675 2790 2848 125 290625 2964 375 30025 3080 625 3138 75 3196875 3255 5313 125 3371 25 3428378 34875 3545 025 3600 75 361 875 2850 5875 25 176 25 235 298 75 3525 41125 470 528 75 5875 64625 705 76375 822 5 88125 940 908 75 1057 5 1116 25 1175 123375 1292 5 1351 25 1410 1468 75 1527 5 1586 25 1645 170875 17625 1821 25 1880 1938 75 1997 5 205625 205 217375 22325 2291 25 2050 2408 75 2467 5 2526 25 2585 264375 27025 2761 25 2820 2878 75 29375 2996 25 3055 311375 31725 3231 25 3090 3348 75 3407 5 3466 25 3525 3583 75 36425 3701 25 2375 59 375 11875 178 125 2375 206 875 356 25 415 625 475 534375 583 75 653 125 7125 771 875 831 25 890 625 950 1009 375 1068 75 1128 125 1187 5 1245 575 1308 25 1365 625 1425 1484 375 1543 75 1803 125 16625 1721 875 1781 25 1840 625 1900 1959375 2018 75 2078 125 21375 2195 875 2256 25 2315 625 2375 2434 375 2493 75 2553 125 26125 2671 875 2731 25 2790 525 2850 2909 375 2068 75 3028 125 9087 5 3148875 3206 25 3265 625 3325 3884 375 3443 75 3503 125 35625 3621 875 3681 25 3740 525 1740 2425 60 625 12125 181 875 2425 303 125 36375 424 375 485 545 625 60625 606 875 7275 788 125 848 7
44. 95 625 51375 571 875 610 644 125 636 25 724375 7825 800 625 838 75 876 875 915 953 125 991 25 1029 375 1067 5 1105 625 1143 75 1181 875 1220 1258 125 1296 25 1334 375 13725 1410 525 1448 75 1486 875 1525 1583 125 1601 25 1630 373 16775 1715625 1753 75 1791 875 1830 1568 125 1906 25 1944 375 19825 2020 625 2058 75 2096 875 2135 2173 125 221125 2249 375 22875 2325 625 230375 2401 875 2015 205375 20925 2181 25 2170 2208 75 2247 5 2286 25 2325 236375 24025 2441 25 1575 38375 7875 116125 1575 196875 23625 275 825 315 354375 35875 433125 4725 511 875 561 25 590 625 630 669 375 70875 748125 7875 826 875 866 25 905 525 945 984 975 1023 75 1063 125 11025 1141875 1181 25 1220 625 1260 1299 375 1338 75 1378 125 14175 1456875 1406 25 1535 625 1575 1614 375 165375 1693 125 17325 1771875 1811 25 1850 625 1890 1929 375 1968 75 2008 125 20475 2088 875 212625 216585 2205 2244375 228375 2323 125 29625 2401 875 2441 25 2480 625 ECM 630 Electroporation System Capacitor uF 1600 1625 1680 4 4085 4125 8125 825 120 121875 12375 180 1625 185 200 203125 206 25 240 24375 205 280 2849 5 28875 320 5 330 36 385825 371 25 400 40625 4125 44 0608 45375 480 487 5 495 520 808 59625 56 s6875 577 5 800 609 375 61875 540 50 860 68 690 675 70125 72 73125 75 760 771875 78375 800 825 840 853105 55 88
45. EQUENTIAL 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 limi tation 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 ECM 630 is changed in any way from its origi nal factory design or if repairs are attempted without written authoriza tion by BTX HARVARD APPARATUS Warranty is void if parts connections or cell fusion chambers not manufactured by BTX HARVARD APPARATUS are used with the ECM 630 Page 2 www btxonline com ECM 630 Electroporation System General Information Continued If a defect arises within the warranty period promptly contact BTX 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 btx harvardapparatus com Goods will not be accepted for return unless an RMA Returned Materials Authorization number has been issued by our customer service department
46. N 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 Page 5 www btxonline com ECM 630 Electroporation System General Safety Summary continued 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 damage to these products or other property Symbols that may appear on the products A 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 Type 20 character by 4 line liquid crystal Display LED backlit 100 to 240 Vae 50 to 60 Hz CAT II 500 W Pulsed 50 W Idle 2 5 A 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 G to C Convection through metal case 60 2 000 m operating 12 5 x 12 25 x 5 5 WxDxH 13 6 Ibs 6 2 kg Single rotary encoder with
47. 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 the 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 WITH SUSPECTED FAILURES If you suspect there is damage to this product have it inspected by qualified service personnel PROVIDE PROPER VENTILATION Refer to installation instructions for details on installing the product to ensure proper ventilation DO NOT OPERATE I
48. d 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 heard 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 Pa
49. ec 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 Sufficiently 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 diver gent 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 per meable 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 membra
50. ectroporation 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 Electrofusion 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 BTX 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 Model Description MAT 45 0001 6300 ECM 630 Electroporation System MA1 45 0051 630 ECM 630 Electroporator only MA1 45 0207 630B Electroporation Safety Stand MA1 45 0124 610 Disposable Electroporation Cuvet
51. ectroporation System 56 25 112 5 168 75 281 25 337 5 39375 506 25 562 5 61875 72125 7875 84375 900 956 25 10125 106875 1125 1181 25 12378 129375 1350 140625 14625 151875 1575 163125 1687 5 174375 1800 1856 25 19125 1968 75 2005 208125 21975 2193 75 2250 2306 25 2382 5 241875 2475 259125 25875 2643 75 2700 275625 28125 286875 2275 56 875 11375 170 625 227 5 284375 34125 388125 455 511875 568 75 625 625 6825 799 375 798 25 853 125 910 966 875 1023 75 1080 625 1375 1194375 1251 25 1308 125 1365 1421 875 1478 75 1535625 1522 5 1549375 1708 25 1763 125 1820 1878 875 193375 1990 625 2047 5 2104375 2181 25 2218 125 2275 2331 875 2388 75 2445 625 2502 5 2559 375 261525 2673 125 2730 2786 875 2843 75 2900 625 2957 5 3014375 3071 25 3128 125 3185 3241875 3208 75 3355825 34125 3468375 3525 25 3583 125 575 115 1725 345 4025 5175 575 6325 747 5 8625 920 977 5 1035 1092 5 1150 12075 13225 1380 14375 1495 15525 1610 1667 5 1725 17825 1840 1897 5 1955 20125 2070 21275 2185 22425 23575 2415 24725 2530 25875 2702 5 2780 2817 5 2875 29325 30475 3105 31625 523 125 581 25 639 975 6875 755 625 81375 871 875 930 988 125 1046 25 1104 375 1625 1220825 127875 1336 875 1395 1453 125 1511 25 1569 375 16225 1685 625 1743 75 1801 875 1860 191
52. er 20 seconds the PULSING ABORTED CHARGING TIMED OUT mes sage 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 properties 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 out lined in Appendix A Page 41 www btxonline com ECM 630 Electroporation System Appendix F Glossary of Electrical Terms Amplitude The instantaneous value of cur
53. latform 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 ECM 630 is the most powerful generator in its class 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 Q 25 Q resolution with none setting Voltage range of 50 V to 2500 V with 5 V resolution and either 25 pF or 50 pF 25 O to 1575 O with 25 O resolution Over 200 ECM 600 protocols may be duplicated with this instrument The additional ECM 630 resistor selection none will allow researchers to reproduce protocols from competitive systems lacking resistor settings or reporting unlimited resistance The ECM 630 will perform the widest range of electroporation applications among commercially available electroporation generators 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 ECM 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 The following items com plete a typical system order Model 630B Electroporation Safety Stand
54. lect 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 ECM 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 screens 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 BTX 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 ECM 630 Reproducing ECM 600 Protocols The ECM 630 will reproduce all ECM 600 protocols 1 Set the voltage as outlined in the ECM 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 100
55. lected 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 ECM 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 pageused to confirm the absence of a pulse 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 aft
56. m 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 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 into the nucleus Applications for electroporation include permeabilization of virtually all cells to a wide variety of molecules and ions The most common appli cations for electroporation are the transformation or transfection of cells with DNA or RNA Other applications for electroporation include electroactivation electroinsertion of proteins into cell
57. 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 Electroporation can be characterized by waveform BTX exponential decay waveform generators such as the BTX ECM 399 and ECM 630 deliver an exponentially decaying pulse The length of such a dis charge 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 capaci tance must be selected on the instrument Voltage may be directly set on the instrument Page 21 Electroporation Process Exponential Decay Wave 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 Elec
58. nes 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 Technical 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 com ponents of competitive systems Please contact BTX Technical Support for details Page 45 www btxonline com ECM 630 El
59. rameters 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 voltage 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 or 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 ECM 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 se
60. re 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 Number aaa 2 CaliDratiOri uie eret tete tr ia 2 ruri 2 3 SERVICE T EO EE 3 4 Repair Facilities and Parts cecessesessseseseeceseeseeeeseeees 4 General Safety Summary es 5 6 Electrical amp Technical Specifications 7 General Specifications eere 8 Introduction ECM 630 Features essent 9 Operation Getting Started 10 Quick Start Installation tie meter tret trcs AA Initializing css Instrument Controls Operating Basics Advanced Operation Programming Electroporation eene rennen 07 Applications Electroporation eerte 22 23 Appendix A ECM 630 Electrode Operation Ranges Appendix B ECM 630 RC Time Constants Appendix C Optimization Strategies Appendix D Electrical Troubleshooting Appendix E Experimental Troubleshooting Appendix F
61. rent 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 membranes of two juxtaposed cells reseal after a DC application Divergence The deviation of electric field lines from a parallel homogeneous con dition 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 cur rents 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 s
62. s 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 A beep is heard when transitioning between LV and HV modes Page 13 www btxonline com ECM 630 Electroporation System Quick Start Continued Resistor 1 2 Resistor indicates the set resistance in Q The resistance range is 25 Q to 1575 Q in 25 0 increments In the LV 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 in microfarads uF In the LV mode the capacitance range is 1UF 25 to 3275 pF in 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 14 The second Set Parameters screen enables the user to save view and load up to three programs Please note that current parameters will 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
63. tes Plus 1mm 50 per bag MA1 45 0125 620 Disposable Electroporation Cuvettes Plus 2 mm 50 per bag MA1 45 0126 640 Disposable Electroporation Page 47 Cuvettes Plus 4 mm 50 per bag 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 Use a soft cloth dampened with water to clean the instrument Use an aqueous solution of 75 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
64. trength 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 at the optimal voltage setting 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 both 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 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 se
65. troporation 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 dyes sugars and dNTP S Electroporation has been used to study cellular activation processes by electropermeabilizing cells to Ca Mg and Na 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
66. w to the left of the parameter 2 To 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 ECM 630 will 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 increment
67. wo 33935 aw 3815 ws 250 260 270 280 200 300 ao 300 am mo 360 seo 265 625 22626 285875 2675 308195 3875 3935 ao 350805 38195 370875 3895 28125 25 90375 316 32625 5 34875 360 371 25 3825 308 75 405 296 675 30875 3200605 25 344075 75625 MO 39875 4075 415605 4275 3125 385 3375 350 3625 375 ws 400 1125 425 4375 450 328 125 34125 354975 3675 380625 a5 405875 4 433125 44625 459 375 ams 34375 3875 s25 385 38875 es 42625 45975 4675 481 25 495 150075 137075 es 416075 5 445625 160 5 48875 5175 375 180 aps 420 ass asp 485 180 ans BU 525 sao 900625 40625 421875 4375 458125 46875 484375 400 51585 59125 546875 5825 40625 4225 438 75 155 0125 487 5 50378 520 538 95 5525 EJ 491875 43875 465 625 425 489 375 50625 523195 540 556875 87375 300 625 6075 4375 465 ams EU sors 525 5425 KO 8775 506 5 30 463126 4126 480379 5 526025 140 76 501 875 s 588 125 1026 633 375 0825 68 26 4875 408 25 506 543 76 662 6 8126 wo 01875 88075 658026 475 484375 50075 523125 5425 561875 58125 565 20 680375 05875 678125 087 5 500 520 540 560 580 00 420 840 680 680 m 720 515625 53525 556875 S775 598125 51875 539375 460 680625 7025 721875 7425 531 25 5525 57375 5 61625 0975 65875 580 70125 7225 743 75 765 546875 36875 560625 8125 504875 5625 sma o 7 855 maia 7656 7875 5825 385 075 630 8525 675 6975 70 7425 765 7875 mo 578 125 60125 654375 ams
68. ystem Quick Start Continued Instrument Controls Pulse Switch ECM 630 ro Cell Mampulato Elect e LI 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 A 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 12 www btxonline com ECM 630 Electroporation System Quick Start Continued Parameter Control Knob The Parameter Control Knob is a rotary encoder controlling 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 arro

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