ECM830多功能型细胞电穿孔 细胞转基因活体导入系统 简介 ...

活体转化显微注射系统技术参数活体转化显微注射系统是活体转化不可缺少的重要仪器，主要用来进行活体转化，包括拉针系统、倒置微分干涉显微镜、显微操作仪与显微注射仪部分组成，可以配合实验需要，满足活体标本的体外转化等实验要求。

其配置需要满足以下技术规格。

打星号的参数为必须满足项目。

设备用途：该设备用于获取清晰的高质量的微分干涉观察效果，可进行显微注射样品角度调整，并用于活体显微注射操作。

技术规格1光学系统部分1.1*无限远光学系统：采用新无限远光学系统，具有轴向和径向双重色差校正，同时具有反差校正，提高图像衬度。

1.2# 45mm国际标准齐焦距离，具备明场，DIC观察功能，可升级PlasDIC功能。

1.3透射光照明器：高亮度长寿命卤素灯照明。

1.4具备智能光源管理功能：可存贮并自动调用各只物镜的最佳照明条件。

1.5光学部件使用金属镀膜，防霉但不得使用化学药剂。

2主机2.1高级显微镜主机，全金属结构，金字塔形主机结构设计，机械温度稳定性高。

2.2调焦系统:组合式金属结构、精细的粗微调焦系统，谐波齿轮设计的防止调焦下滑机构，不得使用易损的摩擦圈作为松紧调节和防止物台下滑机构。

2.36位编码型DIC物镜转盘。

2.4透射/反射切换、照明强度调节可以通过显微镜机身电动钮控制。

2.5# 高光效的V型光路设计2.6观察镜筒：铰链式双目观察筒，金属罩壳，可360度自由旋转，上下翻转；倾斜45度，视野数≥23mm；2.7目镜：放大倍数10x，高眼点，双目屈光度可调，视野数≥23mm。

2.8聚光镜：万能长工作距离聚光镜（可实现相差，微分干涉，塑料皿微分干涉等观察方式），数值孔径0.55，色差球差校正，工作距离≥26mm；2.9微分干涉部件（DIC），有与不同数值孔径的物镜一一对应的棱镜；2.10滤光片转盘孔位≥6，可放置荧光激发块与微分干涉装置；2.11长寿命高亮度荧光照明装置，灯泡寿命≥2000小时，光纤和灯泡耗材满足3年使用消耗。

Gene pulser xcell TM Electroporation system quick guide产品名称: Gene Pulser Xcell 电穿孔系统产品型号: Gene Pulser Xcell 电穿孔系统产品展商: 众磊（北京）生物科技发展有限公司驻沪办简单介绍Gene Pulser Xcell 电穿孔系统Gene Pulser Xcell 电穿孔系统的详细介绍【介绍】电穿孔是功能强大的将核酸、蛋白及其它分子导入多种细胞的高效技术。

通过高强度的电场作用，瞬时提高细胞膜的通透性，从而吸收周围介质中的外源分子。

这种技术可以将核苷酸、DNA 与RNA 、蛋白、糖类、染料及病毒颗粒等导入原核和真核细胞内。

电转化相对其它物理和化学转化方法，是一种有价值和有效的替代方法。

Gene Pulser Xcell 系统的设计，基于Bio-Rad 15 年来在电转化技术上经验积累，它提供指数波和方波波型选择、系统配置选择及友好的用户界面。

主要特点指数波和方波波型确保所有细胞类型（原核及真核）均可获得最佳的电转化效果Bio-Rad 专利的* PulseTrac 电路和电弧保护设计，确保可重复性并保护样品模块化设计可根据研究需要选择系统用户友好的数字化界面，具有直观的编程以控制所有参数，包括附属模块的参数包括人工操作、预设规程、用户规程、一个优化规程及其它先进功能等程序选择指数波或方波脉冲选择Gene Pulser Xcell 系统可产生指数波和方波波型，使你选择最适合你细胞的波型与规程。

指数波和方波均能有效地用于电转化及电融合。

电穿孔波型对不同类型细胞的转化效率有很重要的影响。

左，指数衰变脉冲。

当一个充电至电压为V 0 的电容器放电到细胞，加在细胞上的电压随时间以指数方式下降。

从起始电压下降到V0 / e 所需的时间称为为时间常数τ, 一种方便的脉冲时间表达方式。

右，方波脉冲。

放电到样品后截断电容器脉冲可产生方波脉冲。

电穿孔技术电穿孔缓冲液：配方1：200mM/L葡萄糖,5mM硫酸镁2mM/L疏基乙醇20mM/LTris-HCI ,pH值7.6Universal Electroporation Solution通用电转染缓冲液性能特点：1. 显著提高转染效率和细胞存活率2. 适用于难转染细胞等几乎所有细胞类型3. 与所有电融合/电穿孔仪兼容产品货号：UES0001产品规格：1ml保存：Universal Electroporation Solution保存于4度建议-20度长期保存保质期：正确的使用和保存条件下，保质期为6个月电转染次数：1ml Universal Electroporation Solution用0.4cm电极杯可进行5组实验用0.2cm电极杯可进行10组实验电转染条件：电压260V电容950μF操作步骤：1．收集细胞：用胰酶消化细胞，将细胞培养液吸入到15ml离心管中，1000rpm,5min，弃其上清。

2．用不含血清的培养液洗涤一次，弃其上清。

3．取2～10ug质粒加入到100ul电转缓冲液中，充分混匀。

4．用100ul混有质粒的电转缓冲液充分溶解细胞，转入0.2cm电转导入杯中。

5．将电转导入杯放入样品槽中，释放电脉冲，电击参数按电容1000 mF，电压150-500 V，间隔20V取值，取得最佳效果。

6．立即取出电击杯，分别用一次性吸管将混和液转移至加有完全培养基的一次性细胞培养瓶中，放入细胞培养箱中培养。

电穿孔法转染哺乳动物细胞来源： 发布时间：2009-08-31 查看次数：1935站长注：下文是发表于Nature Methods中的一篇关于电穿孔转染方法的文章，站长对其作了注释，方便大家理解。

电穿孔转染理论上可转染所有的组织细胞，因此对其他如脂质体、磷酸钙沉淀等方法转染效果不明显的细胞可选用此方法。

电转过程中，最重要的就是电穿孔仪的电压、电容以及与电泳缓冲液的选择。

提到电转仪，最出名的恐怕就属BIO-RAD了，他在1986年推出了世界上第一台电穿孔仪，并发布了多种细胞仪电转过程中的电压，电容，电转缓冲液等可省却大家很多的摸索过程，具体资料可到其主页上查找相关Nature Methods 3, 67 - 68 (2006)Transfection of mammalian cells by electroporationPulsed electrical fields can be used to introduce DNA into a wide variety of animal cells1, 2. Electroporation works well with cell lines that are refractive to other techniques, such as calcium phosphate–DNA coprecipitation. But as with other transfection methods, the optimal conditions for electroporation of untested cell lines must be determined experimentally.电穿孔转染可用于磷酸钙转染效率低下的细胞，具有操作简便，转染效率高等优点，但对于不同的细胞，其转染条件有很大的不同，需要进行摸索。

细胞的脂质体转染法和电穿孔转染法（一）脂质体转染一、实验试剂及器材Lipofectamine® 3000 Transfection Reagent，质粒DNA(0.5-5 μg/μL储液)，Opti-MEM®减血清培养基，培养板，酒精灯，离心机，微量移液器，离心管等。

二、实验步骤1. 接种细胞至70－90%汇合度时转染；2. 使用Opti-MEM®减血清培养基稀释Lipofectamine® 3000试剂（2管），充分混匀（6孔板Opti-MEM®培养基为125 μL× 2，Lipofectamine® 3000为3.75和7.5 μL）；3. 在每管已稀释的Lipofectamine® 3000试剂中加入稀释的DNA（用P3000TM试剂稀释）（1:1比例）；4. 室温孵育5分钟；5. 加入DNA－脂质体复合物至细胞中；6. 37°C(PK-15在39°C下培养)孵育细胞2–4天，然后分析转染细胞。

三、注意事项1. 在无血清培养基（如Opti-MEM®减血清培养基）中制备DNA－Lipofectamine® 3000脂质体复合物，直接将其加入含细胞培养基的细胞中（在血清／抗生素存在或不存在时均可）；2. 转染后无需去除转染复合物或者更换／添加培养基；3. Lipofectamine® 3000试剂用量各有不同，测试推荐的两种浓度的Lipofectamine® 3000试剂，以确定最佳用量，开始新的转染；4. Lipofectamine® 3000试剂应放在4℃储存（切勿冷冻）。

附上Lipofectamine 3000 REagent Protocol：（二）电穿孔转染法一、实验试剂及器材BTX ECM2001®2001细胞电融合仪，细胞，胰蛋白酶，电转液，DMEM，培养板，离心机等。

活体电穿孔法介绍1、什么是活体电穿孔活体电穿孔法(in vivo electroporation) 是将外源基因通过电场作用，导入动物目标组织或器官。

由于这种方法能有效导入外源基因，可在多种组织器官上应用,并且效率较高。

活体电穿孔法的原理很简单,在直流电场作用的瞬间,细胞膜表面产生疏水或亲水的微小通道105～115μm ,这种通道能维持几毫秒到几秒,然后自行恢复。

在此期间生物大分子如DNA 可通过这种微小的通道进入细胞。

近年来活体电穿孔法用于转基因研究的报道不断增多,在基因治疗方面的优势也日趋显著,是一种很好的活体基因导入方法。

活体电穿孔法可用于检测瞬时表达系统中载体的表达状况。

大量的研究表明活体电穿孔法在基因治疗方面有非常好的应用前景。

因此目前国内外对活体电穿孔法介导外源基因转移的研究越来越多。

2、活体电穿孔的法的特点活体电穿孔法基因导入和表达效率较高,它的特点主要在以下几个方面:首先,靶器官的选择面广,理论上任何组织和器官都可以作为活体电穿孔的靶器官。

在用于基因治疗方面，要考虑到靶器官组织生理特性。

如果所选择的局部组织细胞不能把所转移基因的表达产物分泌到外周血液循环中,则在某种意义上说已失去了基因导入的价值,这在基因治疗中是关键性的问题。

当使用组织特异性表达载体时,研究人员应根据所构建的表达载体来选择基因转移和表达的靶器官组织。

例如鱼精蛋白21 启动子可指导外源基因在精母细胞中特异性表达,以小鼠的睾丸作为靶器官将含有鱼精蛋白21启动子的表达载体导入,获得外源基因的表达量远远高于该基因在肝脏和骨骼肌中的表达。

其次,对导入的外源基因片段的大小没有限制,从几KB 或十几KB 的表达载体, 到100～200KB 的YAC、BAC 基因组,都有成功导入并获得表达的报道。

此外活体电穿孔法操作简单快速,电穿孔的时间只有几秒钟,而且DNA片段不需要特殊的纯化操作。

但电穿孔法也存在一些的缺点:首先,外源基因表达持续的时间很短,虽然外源基因导入后最快可在215 小时有表达,但大多1～2 月后表达量降至很低。

ECM ® 830Instruction ManualWarrantyThe BTX Instrument Division of Genetronics, Inc. (Genetronics) warrants that the ECM 830 is free of defects for a period of two (2) years from time of delivery from an authorized Genetronics Distributor. If any defects covered by this warranty appear within the above period, Genetronics shall have the option of repairing or replac-ing the equipment at its expense. Such repair or replace-ment shall be the customer’s exclusive remedy for breach of warranty or for negligence. This warranty does not extend to any instrumentation which has been (a) sub-jected to misuse, neglect, accident or abuse, (b) repaired or altered by anyone other than Genetronics without Genetronics’ express and prior approval, (c) used in violation of instructions furnished by the BTX Instrument Division of Genetronics, Inc. Manufacturer shall not be liable for any special or consequential damages or for loss, damage or expense (whether or not caused by orresulting from Manufacturer’s negligence) directly or indirectly arising from use of the instrumentation sold hereunder either separately or in combination with any other equipment or from any other cause.The above warranty shall be in lieu of and excludes all other expressed or implied warranties of merchantability, or fitness for any purpose, or otherwise. Without limiting the generality of the foregoing, Manufacturer 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.Manufacturer will correct any malfunction not caused by operator abuse at no charge for parts and labor. All service under the warranty will be made at the Genetronics, Inc. San Diego, CA facilities or at another location approved by Genetronics, Inc. Owner will ship instrument prepaid to San Diego, CA. Manufacturer will return the instrument after servicing, freight prepaid to owner’s address.Warranty is VOID if the instrument is changed in any way from its original factory design or if repairs are attempted without written authorization by Manufacturer.Warranty is VOID if parts or connections not manufac-tured by Manufacturer are used with a BTX instrument.Note: under no conditions should the instrument or accesso-Table of ContentsWarranty Information (i)Customer and Technical Service Information (2)Safety Guidelines (3)Safety Terms and Symbols (4)Electrical and Technical Specifications (4)Getting Started (7)Features (7)Quick Start (7)Connecting (7)Initializing (8)Instrument Controls (9)Operating Basics (16)Advanced Operation (17)Electroporation/Electrofusion Theory (19)Electroporation (19)Electrofusion (20)Applications (21)Electroporation (21)Bacteria and Yeast (21)Mammalian Cells (21)Plant (22)Other (22)Electrofusion (23)Embryo Manipulation (23)Hybridoma Formation (23)Plant Protoplast (23)Other (23)AppendicesAppendix A:ECM 830 Electrode Operation Ranges (24)Appendix B:Optimization Strategies (28)Appendix C:Electrical Troubleshooting (30)Appendix D:Experimental Troubleshooting (30)Appendix E:Glossary of Electrical Terms (32)Appendix F:Glossary of Biological and Technical Terms (33)Appendix G:Electroporation and Electrofusion Pulse GeneratorCompatibility (36)Appendix H:Recommended Reading (37)Appendix I:Product Information and Ordering (37)Appendix J:General Care and Cleaning (38)Appendix K:BTX Generator Footswitch Instructions (39)NotesPage 2Customer and Technical ServiceInformationCustomer ServiceFor any inquiry or request for repair service, contact the BTX Instrument Division of Genetronics, Inc. Customer Service group in writing or by the following:Phone: 1-800-289-2465 1-858-597-6006Fax: 1-858-597-9594E-mail: cust@If outside the United States and Canada, call: 1-858-597-6006or contact your nearest BTX Distributor.Technical ServiceBTX is the ultimate resource for technical information on the use of high voltage electric fields for performing highefficiency cell fusion, embryo manipulation, gene transfer,bacterial transformation and general electroporation ofmolecules and drugs into cells. We constantly track andmonitor all scientific publications in this area. Our Technical Service group extracts and enters pertinent information, such as results and parameters from these papers into a Database Management System. The resultant Electronic Genetics®Database can be accessed and searched on any combination of the field identifiers.For technical assistance or information, contact the BTX Instrument Division of Genetronics, Inc. Technical Service group:Phone: 1-800-289-2465 1-858-597-6006Fax: 1-858-597-9594E-mail: tech@If outside the United States and Canada, call: 1-858-597-6006or contact your nearest BTX Distributor.BTX ®A Division of Genetronics, Inc.11199 Sorrento Valley RoadSan Diego, CA 92121-1334U.S.A.Page 3830 ManualSafety GuidelinesReview 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 asspecified.Only qualified personnel should perform service procedures.To Avoid Fire or Personal InjuryUse Proper Power Cord. Use only the power cordspecified for this product and certified for the country of use.Connect and Disconnect Properly. Do not connect ordisconnect 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 avoidelectric shock, the grounding conductor must be connected to earth ground. Before making connections to the input or 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 ratingspecified for this product.Avoid Exposure to Circuitry. Do not touch exposedconnections and components when power is present.Do Not Operate With Suspected Failures. If you suspect there is damage to this product, have it inspected byqualified service personnel.Provide Proper Ventilation. Refer to installation instruc-tions for details on installing the product to ensure proper ventilation.Do Not Operate in Wet/Damp Conditions.Do Not Operate in an Explosive Atmosphere.Keep Product Surfaces Clean and Dry.Safety Terms and SymbolsThese terms may appear in this manual:WARNING – Warning statements identify conditionsor practices that could result in injury orloss of life.CAUTION – Caution statements identify conditions orpractices that could result in damage tothis product or other property.These terms may appear on the product:DANGER – Danger indicates an injury hazardimmediately accessible as you read themarking.WARNING – Warning indicates an injury hazard notimmediately accessible as you read themarking.CAUTION – Caution indicates a hazard to propertyincluding the product.Electrical and Technical SpecificationsElectrical and Technical Specifications DisplayType: 20-character by 4-line liquid crystal LED backlitPower SourceV oltage: 100 – 240 Vac, 50 - 60 Hz, CAT IIPower: Pulse: 500WIdle:150WFusing: T2.5250VEnvironmentalFor Indoor Use OnlyOperating Temperature: 10º C to + 40º CCooling: Convection through metal caseHumidity: ≤ 90% relative humidity830 ManualMechanicalFootprint: 12.5” x 12.25” x 5.5” (31.75 x 31.12 x 13.97 cm) (W-D-H)Weight: 15 lbs (6.8 kg)Controls: Single rotary encoder with integrated push buttonElectricalVoltage5-500 V olts (LV mode) @ 1 V olt resolution30-3000 V olts (HV mode) @ 5 V olts resolution(Voltage Delivery @ 5% accuracy and Voltage Monitoring @ 5.0% accuracy)Pulse Length10µs - 999µs (LV mode) @ 1µs resolution1ms – 999ms (LV mode) @ 1ms resolution1s - 10s (LV Mode) @ 0.1 s resolution10µs - 600µs (HV mode) @ 1µs resolutionPulse Interval100ms – 999ms @ 1 ms resolution1.0s – 10.0s @ 0.1s resolution (upper limit is power limited)Multiple Pulsing1 – 99 pulsesCurrent Limit500A limit at 100µsPollution Degree 2Not to be operated in conductive pollutants atmosphere* Note: See Side BarGeneral SpecificationsCertifications and Compliance’sOver voltage Category:CA T III Products in this Category: Distribution-level mains, fixed installationCA T II: Local-level mains, applications and portableequipmentCA T I: Signal levels in special equipment or parts ofequipment, telecommunications and electronicsMeets requirements of Directive 89/336/EEC for Electromag-netic Compatibility (EC) and Low-V oltage Directive73/23/EEC for Product Safety.Compliance was demonstrated to the following specifications as listed in the Official Journal of the European Communities: EN 50081-1 EmissionsEN 55011 Class B Radiated and Conducted EmissionsEN 55082-1 ImmunityIEC 10004-2 Electrostatic Discharge ImmunityIEC 10004-3 RF Electromagnetic Field ImmunityIEC 10004-4 Electrical Fast Transient/Burst ImmunityLow V oltage Directive 73/23/EECEN 61010-1 Safety requirements for electrical equip-ment for measurement, control and laboratory use.830 ManualGetting StartedCarefully open the box containing the ECM 830 Electropora-tion system. Verify receipt of the following items:ECM 830 Pulse Generator (1)Power Cord (1)BTX Electroporation Systems may be customized with the addition of various electrodes and accessories. Thefollowing items complete a typical system order:Model 630 B 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, pleaseverify their receipt.FeaturesThe ECM 830 is a Square Wave Electroporation Systemdesigned for all in vitro and in vivo electroporation applica-tions. The generator utilizes the new BTX Power Platform Technology and an all-new digital user interface.Quick StartConnectingInsert female end of power cord into male power interface on the back panel of the ECM 830. Plug male end of power cord into appropriate electrical outlet. Insert male banana plugs of the 630 B Safety Stand or alternative electrode device into High V oltage output located on front panel of the ECM 830.InitializingPush the power switch located on the front panel of the ECM 830. The display will flash:ECM 830ElectroporatorOS=v1.08, App =v0.00Genetronics, Inc.Following this initialization screen, the first time the instru-ment is initialized, the factory default display will then read:Set ParametersMode: LVV oltage: 0050VP. Length: 020 msThe first page of the Set Parameter Screen from the last time the ECM 830 was used will be displayed each time the ECM 830 is powered up after this initial startup.830 Manual ParameterControlPowerSwitchParameter Control KnobThe Parameter Control Knob is a rotary encoder controlling both the parameter (i.e. voltage) and the value of the param-eter under control (i.e. 50 volts). The display will indicate which parameter is under control by the presence of an arrow to the left of the parameter. 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 left of the value displayed for that parameter. To adjust the value of a parameter under control, rotate the knob clockwise to increase the value and counter clockwise to decrease it. 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. In order to move to the previous screen, move the cursor to the top of the screen and rotate the knob counterclockwise.DisplayThe ECM 830 Display will show two possible screens, the Set Parameter Screen and the Pulsing Completed Screen as well as various Status Messages.Set Parameter ScreenThe Set Parameter Screen is indicated by a display showing “Set Parameters” on the first line of each of three screen pages. The Mode, V oltage and Pulse Length are shown on the first page; the Number of Pulses, Pulsing Interval and Polarity are shown on the second page and the Advanced Features including Save, View and Load are shown on the third and final page of the Set Parameter Screen. The ECM 830 will beep when toggling from one page to another. ModeThe Mode indicates “LV” for low voltage mode or “HV” for high voltage mode. Adjusting the voltage as appropriate automatically controls the mode. There is no manual mode control.VoltageThe V oltage indicates the set voltage in volts. The low voltage mode range is 5V– 500V in 1V increments and the high voltage mode range is 30V-3000V in 5V increments. A beep is heard when transitioning between LV and HV modes.830 ManualTable 1P . LengthPulse Length indicates the set pulse length in m s or ms. The pulse length minimum is 10 m s in both HV and LV modes. In the LV mode, the maximum pulse length is 10 seconds. The pulse length units change from m s to ms at 1ms and from ms to s at 1s. Refer to Table 1 for HV pulse length maximum limits. To estimate approximate pulse lengths at various voltages in the high voltage mode, refer to Table 1 below.Please note that the voltage determines the maximum pulse length in the HV mode. If the voltage is increased and the pre-set pulse length is longer than the maximum pulse length allowed, the pulse length will automatically adjust to that level.Page 12Interval protocols!Programability protocols # Pulses The # Pulses indicates the number of set pulses from 1-99.Interval The Interval indicates the time duration between pulses. The interval range is 100 ms to 10.0 seconds, switching from units of ms to seconds after 999 ms, with 0.1 sec resolution.Advanced Features The final Set Parameters page enables the user to save, view, and load up to three programs. Please note that parameters can be changed inadvertently 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 instant use.Save 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 new screen is displayed. The new screen will read Saved Current Parameters to Set N . Push the parameter control knob again to return to the setup screen.View To view the appropriate program parameters, push the parameter control knob to move the arrow to the left of the program number. Select the appropriate program number by rotating the parameter control knob, then hold the knob in, releasing after a new screen is displayed. The new screen will outline the parameters currently stored under that program number. Push the parameter control knob again to return to the setup screen.Load 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. Rotate the knob to change the program number. Push and hold the knob in, releasing after a new screen is displayed. The new screen will read Loaded Set N to Current Parameters . Push the parameter control knob again to return to the setup screen.On-Line Help In the Set Parameter Mode, an On-Line Help function is enabled. On-Line Help provides a definition for all setparameters and advanced functions. To use On-Line Help,830 Manual 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). Push the parameter control knob in and hold until the definition is displayed. Push a second time to return to the Set Parameter Screen.ModeLow voltage or high voltage range modeVoltageThe amplitude of the output pulseP LengthThe time duration or length of the pulse# PulsesThe number of output pulsesIntervalThe time interval between pulsesSaveSave current setup parameters to nonvolatile memoryViewShow the setup parameters stored in nonvolatile memoryLoadLoad setup parameters from memory to useStatus MessagesFollowing the initiation of a pulsing sequence, various status messages are displayed. The following status messages may be observed: Charging, Pulsing, Pulse Aborted, and Tem-perature Failure.ChargingAs soon as the start switch is pressed, the Charging status message is displayed. The capacitors in the ECM 830 are overcharged during the duration of this message.PulsingOnce the capacitors have reached the preset voltage level, they are discharged and the Pulsing status message is displayed for the duration of the pulse or preset number of pulses. A click or a beep will be heard upon the delivery of each pulse. The sound that is heard is a function of the pulse length. At pulse lengths less than 1-2 ms, clicks will likely be heard. At pulse lengths at or above 1-2 ms beeps should be heard. A final, more extended beep will be heard upon completion of the pulsing sequence. Following the pulsing status message, the Pulsing Completed Screen is displayed.Pulsing Aborted During ChargingIf the start switch is activated a second time prior to delivery of a pulse, the pulsing sequence is aborted and the Pulsing Aborted During Charging status message is displayed.Temperature FailureIf unit is rapidly pulsed at the high end of the two voltage modes, it is possible for a high temperature warning to occur. In this event, the status message observed is Unit Cooling Please Wait . When the unit has cooled down to a safe level, two beeps will sound and the message will change to Ready . Pushing the start button, or rotating the parameter control knob will shift user back to Parameter Set Screen.Pulsing Completed ScreenThe Pulsing Completed Screen is displayed following the delivery of a pulse or train of pulses and is indicated by the display Pulsing Completed on the top line of each of two screen pages. Page 1 comes up automatically; rotate the parameter control knob clockwise to view page 2 and counterclockwise to go back to page 1. Press the knob to return to the Set Parameters Screen or press the start switch to deliver another pulse. The pulsing completed screen displays monitored parameters including Mode, V oltage, Pulse Length, Number of Pulses, Pulse Interval and Polarity. Please note that parameters will be displayed, even if Safety Stand or other output is not connected. Always verify that your sample is connected with the HV output.ModeMode indicates the voltage mode used in the delivery of the last pulse: LV or HV.830 Manual VoltageV oltage indicates the output voltage of the last pulse, in volts. For pulses of 10 255 µs, the voltage is measured at the end of the pulse, and for pulses longer than 255 µs, the voltage is measured at the beginning, or peak.P LengthP Length indicates the duration of the pulse in m s from10-999 µs, ms from 001 to 999 ms, or in seconds from 1.0 to 10.0 s.# Pulses# Pulses delivered in the last pulsing sequence is displayed, from 1-99.IntervalThe Interval between the next-to-last and the last pulse is monitored and displayed in ms from 100-999 ms and in seconds from 1.0 to 10.0 seconds. If only one pulse is delivered, none is displayed.PolarityThe Polarity used in the last sequence of pulses is monitored and displayed as Unipolar.High Voltage OutputThe High V oltage Output is located in the lower center portion of the front panel. Plug the high voltage cables into this output following the color-coded polarity.SmartLinkThe Smartlink port on v1.08 ECM 830 and beyond is equipped with footswitch connection capabilities. If your instrument falls in this category and you wish to use the footswitch, please see appendix K on page 39. If your instrument does not fall into this category, contact BTX customer service at (800) 289-2465 orcust@ for assistance.630B & 630 RB-1Simple, single dial operation with parameter control knobOperating BasicsUse with Safety Stand 630B and BTX Disposable Electroporation Cuvettes Plus1.Insert the safety stand banana plugs into the HV Output on the front panel of the ECM 830.2.Press the power switch to initialize the ECM 830.3.Rotate parameter control knob to move arrow to voltage and push to select. Rotate knob to adjust voltage. Push again to select that voltage.4.Rotate parameter control knob to move arrow to P Length and push to select. Rotate knob to adjust pulse length. Push again to select that pulse length.5.Rotate parameter control knob to move arrow to #Pulses (Page 2) and push to select. Rotate knob to adjust the number of pulses. Push again to select that number of pulses.6.Rotate parameter control knob to move arrow to Interval if you have designated more than one pulse in 5 and push to select. Rotate knob to adjust the pulse interval. Push again to select that pulse interval.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 830 will charge and then deliver the electroporation pulse or train of pulses selected, beeping with the delivery of each pulse.9.Process sample. Do not forget to recordappropriate parameters as displayed on thePulsing Completed Screens, for documentationpurposes.10.To return to ready mode, press the Start button.11.To abort a pulse before delivery, press the Startbutton during the charging mode.Use with Alternative Electrode Chambers and ApplicatorsThe ECM 830 may be used to power all BTX electroporation and electrofusion chambers and applicators. 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 830.Use with FootswitchPlease see Appendix K on page 39 Advanced Operation: Programming1.Set parameters as outlined in Operating Basics.2.Once presetting of parameters has been completed, rotatethe parameter control knob until the save function has been reached.3.Press knob in and release. Now rotate to select whichprogram number the chosen settings (see Preset Param-eters above) should be saved under. Program #1 isreserved for current parameters in active use and cannotbe used for pre-set storage.4.Push and hold knob in, releasing only after a new screenreading Saved Current Parameters to Set N is displayed.(N refers to program number designated in step 2)5.Push the parameter control knob again to return to the SetParameter Screen.6.From the Set Parameter Screen, use the parametercontrol knob to rotate until the load function has beenreached.7.Push the knob in and release. Now rotate to select theappropriate program number.8.Push and hold knob in, releasing only after a new screenreading Loaded Set N to Current Parameters isdisplayed.9.Push the parameter control knob again to return to theSet Parameter Screen.Pore FormationElectroporation ProcessElectroporation/Electrofusion TheoryElectroporationElectroporation 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 mem-brane. The pores formed are of the order of 40-120 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; forexample, 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 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 aresearch tool, recent work has demonstrated its potential for clinical applications. Some areas being exploredinclude: electrochemotherapy which involves electropora-tion 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.Electrofusiontransgenic plant production. characterized by waveform.Voltage may be directly set on the instrument. gentler to cells which yields higher viability.Alternating CurrentMammalian Cells Expressing GFP Following EPAlternating current generators, such as the BTX ECM 2001and radio frequency (RF) generators deliver a sinusoidal wave. These are used to cause dielectrophoresis, in which cells may be brought together for fusion. Parameters, which typically may be set, include amplitude, duration and frequency.ApplicationsElectroporationBacteria and Yeast ElectroporationThe most common application is transformation. Fieldstrength and pulse length are critical parameters for reporting,optimization and troubleshooting bacterial and yeast applica-tions.Mammalian Cell ElectroporationElectroporation has been used successfully to introduce many different molecule types into cells. Most commonly, elec-troporation 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 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 electroincorporated. 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 2+, Mg 2+ 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 bleomy-cin, 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 param-eter may be very much dependent on the impedance of the sample (sample load).。

MicroPulser电穿孔仪操作手册2018年12月27日1、介绍（1）基本原理MicroPulser电穿孔仪用于细菌、酵母和其他众多微生物的电击转化，转化时，高压电脉冲作用于悬浮在小体积高阻介质中的样品。

本系统由一个脉冲发生器（pulse generator）模块、一个电击腔（shocking chamber）和一个装有电极的电击杯（cuvette）组成。

样本放置于电击杯的电极之间。

MicroPulser模块包含一个电容器，将电容器充电至高电压，然后模块将电容器中的电流放电到试管中的样品中。

MicroPulser的电容放电电路产生具有指数衰减波形的电脉冲，如下图。

当电容器放电至样品时，跨越电极的电压迅速上升至最大电压（or峰值电压，peak voltage；也称为初始电压，Vo），并随时间（t）减小，如下式：其中τ=R·C，为时间常数，是脉冲长度的简便表达式。

R为电路电阻，单位为ohms（欧姆）。

C为电容，单位为microfarad（微法拉）。

根据方程1，τ是电压下降至峰值电压1/e（~37%）的时间。

MicroPulser的内部电路被设计以使E.coli、酿酒酵母及其他许多微生物可以得到最佳电穿孔，最佳转化效率发生在大约5ms的时间常数内。

这些电穿孔条件是通过使用10微法拉电容器和将600欧姆电阻与样品池并联以及将30欧姆电阻与样品池串联来实现的。

除时间常数外，电场强度是另一个决定转化效率的重要参数。

电场强度E，是施加于电极间的电压，公式为：其中，V为施加的电压，d为电极间的距离，单位为cm。

电场强度和细胞的尺寸（size）决定了横贯每个细胞的电压降，正是电压降可能是电穿孔中电压效应的重要表现。

30欧姆串联电阻的目的是在发生电弧的情况下保护设备电路。

在正常操作条件下，当样本在高电阻介质中，电阻不会影响施加在样本上的电压。

但是，当样本的电阻较低时，电阻将极大地降低施加在样品上的电压。

一、酵母菌电转化方案:方案一。

1、负80度取出涂平板。

30度培养二天。

2、取平板单菌落接种到10ml YPD培养基的50ml的三角瓶中30度过夜摇培3、4度3000~4000rpm半分钟离心收获细胞。

4、用20ml TE/LioAC(0.1M/0.1M pH7.5)重悬细胞，并在30度摇床摇培45分钟5、加0。

5ml 1M DTT 再摇15分钟6、加20ml双蒸水4度3500rpm离心半分钟第一次沉淀用30ml冰冷的无菌水重悬第二次沉淀用20ml冰冷的1M 的山梨醇重悬第三次沉淀用1ml冰冷的1M 的山梨醇重悬8、每40ul一个转化体系分装9、转化条件1。

5KV 20uF 200Ω 40ul yeast /reaction 100ng DNA/reaction<5ul10、加150ul 1M山梨醇快速混合酵母和DNA，然后涂在选择培养基上。

方案二、1、负80度取出涂平板。

30度培养二天。

2、取平板单菌落接种到10ml YPSD（加SORBITOL）培养基的50ml的三角瓶中30度过夜摇培3、4度3000~4000rpm半分钟离心收获细胞。

4、加20ml双蒸水4度3500rpm离心半分钟5、第一次沉淀用30ml冰冷的无菌水重悬第二次沉淀用20ml冰冷的1M 的山梨醇重悬第三次沉淀用1ml冰冷的1M 的山梨醇重悬6、每40ul一个转化体系分装7、转化条件1。

5KV 20uF 200Ω 40ul yeast /reaction 100ng DNA/reaction<5ul8、加1000ul 1M山梨醇快速混合酵母和DNA，9、30度温浴1~2小时，不能摇动。

然后涂在选择培养基上。

二、怎么样选择电转仪（电转仪的功能与适用对象）美国著名BTX是专业的细胞融合、电穿孔仪的生产厂家。

自从1983年起，苛求的科研工作者就已经把BTX产品作为电融合、转基因等应用领域的首选仪器。

除了占世界领导地位的电融合、转基因系统外，BTX的产品还包括多达25种相关的配件，其中有多种专业的电极，给用户更多的选择。

一、酵母菌电转化方案:方案一。

1、负80度取出涂平板。

30度培养二天。

2、取平板单菌落接种到10ml YPD培养基的50ml的三角瓶中 30度过夜摇培3、4度3000~4000rpm半分钟离心收获细胞。

4、用20ml TE/LioAC(0.1M/0.1M pH7.5)重悬细胞，并在30度摇床摇培45分钟5、加0。

5ml 1M DTT 再摇15分钟6、加20ml双蒸水 4度3500rpm离心半分钟第一次沉淀用30ml冰冷的无菌水重悬第二次沉淀用 20ml冰冷的1M 的山梨醇重悬第三次沉淀用 1ml冰冷的1M 的山梨醇重悬8、每40ul一个转化体系分装9、转化条件1。

5KV 20uF 200Ω 40ul yeast /reaction 100ngDNA/reaction<5ul10、加150ul 1M山梨醇快速混合酵母和DNA，然后涂在选择培养基上。

方案二、1、负80度取出涂平板。

30度培养二天。

2、取平板单菌落接种到10ml YPSD（加SORBITOL）培养基的50ml的三角瓶中 30度过夜摇培3、4度3000~4000rpm半分钟离心收获细胞。

4、加20ml双蒸水 4度3500rpm离心半分钟5、第一次沉淀用30ml冰冷的无菌水重悬第二次沉淀用 20ml冰冷的1M 的山梨醇重悬第三次沉淀用 1ml冰冷的1M 的山梨醇重悬6、每40ul一个转化体系分装7、转化条件1。

5KV 20uF 200Ω 40ul yeast /reaction 100ngDNA/reaction<5ul8、加1000ul 1M山梨醇快速混合酵母和DNA，9、30度温浴1~2小时，不能摇动。

然后涂在选择培养基上。

二、怎么样选择电转仪（电转仪的功能与适用对象）美国著名BTX是专业的细胞融合、电穿孔仪的生产厂家。

自从1983年起，苛求的科研工作者就已经把BTX产品作为电融合、转基因等应用领域的首选仪器。

除了占世界领导地位的电融合、转基因系统外，BTX的产品还包括多达25种相关的配件，其中有多种专业的电极，给用户更多的选择。

Btx ecm2001电转染参数【导言】Btx ecm2001电转染系统是一种常用的实验仪器，广泛应用于生物医学研究领域。

在使用这一设备进行电转染实验时，合理的操作参数设置对实验结果的准确性和稳定性有着重要的影响。

本文将对Btxecm2001电转染系统的参数设置进行详细的分析和介绍，以帮助研究人员更好地使用这一设备开展电转染实验。

【主要内容】1. 电转染系统简介Btx ecm2001电转染系统是一种常用的电转染仪器，主要用于将DNA、RNA、蛋白质等生物大分子转移到细胞内，从而实现基因转染、蛋白质转染等实验目的。

该系统具有电极板、电源控制器、电解质缓冲液等组成部分，能够提供稳定的电场和温度环境。

2. 电转染参数设置在使用Btx ecm2001电转染系统进行实验时，需合理设置以下参数：（1）电压：电压的选择要根据具体实验需求而定，一般可在50-300V范围内进行调节。

较低的电压可以减少细胞膜的破坏，但转染效率较低；较高的电压能够提高转染效率，但易引起细胞膜的损伤。

因此需根据实验要求进行合理选择。

（2）时间：电转染时间也是影响转染效率的重要参数。

通常电转染时间在25-40分钟之间为宜，过长或过短的时间都可能影响转染效果。

（3）电极间距：电极间距的选择与所用细胞培养皿的尺寸有关。

通常情况下，电极板与培养皿的距离应控制在1-2mm范围内。

（4）电解质缓冲液：电解质缓冲液的配制和选择也对转染效果产生影响。

常用的电解质缓冲液有PBS、HEPES等，需根据实验需要选择合适的缓冲液。

3. 注意事项在进行电转染实验时，还需注意以下事项：（1）操作规范：操作时需严格按照设备说明书和实验流程进行，以确保实验的准确性和安全性。

（2）细胞预处理：在进行电转染实验前，需对细胞进行合适的预处理，包括细胞的培养、细胞数目的调整等。

（3）质控实验：在进行正式实验前，可进行一定数量的质控实验，以验证实验条件的合适性。

4. 实验结果分析在实验完成后，需对实验结果进行详细的分析和比对。

活体电穿孔法介绍活体电穿孔法（in vivo electroporation）是一种利用电场作用实现基因传递的技术方法。

它通过向生物体的细胞施加较高电场脉冲，使细胞膜发生破坏和通透性增加，从而使外源基因能够进入细胞内。

活体电穿孔法广泛应用于基因治疗、基因转导、细胞修复等领域，在生物医学研究和临床实践中具有重要的应用价值。

活体电穿孔法的基本原理是利用电场作用来改变细胞膜的通透性。

在传统的基因转导方法中，如病毒载体介导的转染，局限性较大，容易受到免疫反应和基因表达的不稳定性等问题。

而活体电穿孔法则可以针对不同细胞类型和组织进行精确控制，实现高效、可重复的外源基因转导。

电场脉冲的产生是活体电穿孔法的前提条件。

通常使用电脉冲发生器来产生足够高的电场脉冲。

电参数的选择是关键，包括脉冲的电压、持续时间、频率等。

这些参数的选择需要根据细胞类型、组织状态以及具体实验目的来确定。

电穿孔过程是指电场作用下，细胞膜发生通透性增加和破坏的过程。

电场脉冲会导致细胞膜上的孔洞形成，从而使外源基因能够进入细胞内。

这个过程主要涉及细胞膜的电极化和去电极化。

电极化是指细胞膜在电场作用下的电位变化，去电极化则是指细胞膜恢复到正常状态的过程。

基因传递与表达是活体电穿孔法的最终目的。

一旦细胞膜被电穿孔，外源基因就有机会进入细胞内，并在细胞内进行表达。

通常在电穿孔后立即给予外源基因的加入。

外源基因可以是DNA、RNA等，具体选择要根据研究需要来确定。

通过活体电穿孔法传递的基因可以实现细胞的基因修复、基因敲除、基因转录调控等。

活体电穿孔法的应用范围十分广泛。

在基因治疗方面，活体电穿孔法可以用于将修复基因传递到患者体内，治疗一些遗传性疾病。

在基因转导方面，活体电穿孔法可以用于将外源基因转导到特定细胞或组织中，实现特定功能的表达。

在细胞修复方面，活体电穿孔法可以用于修复受损细胞的基因，促进组织再生与修复。

总结来说，活体电穿孔法是一种基于电场作用的基因传递技术。

基因导入仪电穿孔仪安全操作及保养规程1. 前言基因导入仪电穿孔仪是一种用于细胞转染的设备，通过给予细胞电压，将要导入的基因或其他物质导入到细胞内，是分子生物学研究中不可或缺的工具之一。

然而，如何正确和安全地操作电穿孔仪，以及如何保养设备，是非常重要的问题。

以下是基因导入仪电穿孔仪的安全操作及保养规程。

2. 安全操作规程2.1 立场规范使用人员必须端正态度，认真工作，严格遵守操作规程，保证性能稳定，设备正常工作。

操作过程中，切勿手动给设备增加额外电源，对操作面板、开关按钮及插头的操作，需轻拿轻放。

设备进水或发生异常时，应立即停止开机、切断电源，报告管理员进行检查和维修。

2.2 有关电源的注意事项基因导入仪电穿孔仪应只能在配套电源下进行开机，禁止任何行为耗用或使用非公司配套电源。

使用设备时，应先确认电源插头与插座匹配，确认好不松动插好后接通电源。

操作过程中，建议使用三线插座加接地线，将设备接地。

禁止在设备内部随意接线、插线，禁止乱动插头，禁止连接使用已经损坏、老化的连接线、插头。

操作人员切勿随意移动电源，禁止在设备配线板及上方空间敷设电线，应保持配线整齐，避免扭结。

在操作时，设备前侧应留有足够的工作距离，操作过程中切勿投入危险区域，切勿迫近放置在培养皿中的细胞。

2.3 操作环境的要求在室内操作时，腰板之间应保持一定距离，操作过程中应避免口述、讲话，不得使用高频器、超音波打泡器及其它电动仪器等引起干扰的器材。

要选择放有平行电场的工作台，以保证正常操作，防止操作错误。

在操作过程中，应运用杀菌器、蒸汽消毒器等先进设备，确保培养器具和各种操作表面的清洁和消毒。

切勿将污染的培养器具、材料放入工作台中，以保证操作环境和细胞的纯度。

2.4 操作人员的要求合法持证上岗的操作人员，无论非常忙或有事时，都切勿在操作时急躁、浮躁。

切勿批量进行基因导入实验，以防细胞繁殖异常，影响实验结果。

为防止误操作，须事先确认自己的能力和经验能否满足实验要求。

免疫细胞转染原理免疫细胞转染是指将外源的DNA、RNA、蛋白质等分子转移到免疫细胞内，使其表达或产生特定的功能。

免疫细胞转染通常采用基因转染技术。

常见的方法有病毒载体转染、电穿孔、化学转染等。

病毒载体转染是最常用的免疫细胞转染方法之一。

常用的载体有腺病毒、逆转录病毒等。

将带有目标基因的病毒载体加入到免疫细胞培养液中，病毒侵入免疫细胞，并释放出目标基因。

然后目标基因会被免疫细胞的细胞核摄取并表达。

电穿孔是通过电脉冲破坏细胞膜，形成暂时的微孔，使外源基因能够进入免疫细胞内。

通常使用特定的设备和电参数来实现电穿孔。

化学转染则是通过化学试剂促进外源基因进入免疫细胞。

化学试剂可以改变细胞膜的渗透性，使外源基因能够穿越细胞膜进入细胞。

免疫细胞转染的原理是通过上述方法将目标基因转入免疫细胞内，从而改变免疫细胞的遗传信息，使其表达或产生特定的功能。

这对于研究免疫细胞的功能或治疗免疫相关疾病具有重要意义。

除了上述提到的转染方法，还有一些其他常用的免疫细胞转染方法：1. Liposome转染：使用具有正电荷的脂质体（liposome）来包裹目标基因，通过静电相互作用与免疫细胞膜结合，从而促进基因转入细胞内。

2. 基因枪转染：将高压气体通过准点射出的形式，直接将基因载体瞬间推进免疫细胞内，以实现基因转染。

3. 发射武器转染：使用强电场或激光脉冲等方式，将基因载体通过微小的发射枪射入免疫细胞。

这些方法均能够将外源基因引入免疫细胞内，在其中进行表达或产生特定的功能。

转染前需要选择适当的转染载体和方法，以确保高效率和低细胞毒性。

免疫细胞转染的应用广泛，如基因疗法、细胞治疗、疫苗研发等。

通过转染，可以改变免疫细胞的表型和功能，从而增强免疫应答或治疗免疫相关疾病。

细胞电融合仪ECM?2001安装/操作手册目录1．检查清点货物及安装2．技术规格3．操作细则4．产生杂交瘤的实验方法5．服务及保修6．附录：电穿孔技术在转基因及动物克隆中的应用第一节检查清点货物及安装1.拆封包装：ECM 2001细胞电融合仪采用纸箱包装，收到货物后，请检查包装完好程度，如果有任何损伤请速与我公司联系。

请小心地拆开包装，将仪器及附件取出，并依据合同清点货物内容及数量，如果有任何外观损伤或者货物与合同有差异，请速与我公司联系。

请保留包装箱，以便满足将来一旦要运送该仪器的需求。

2.电源：该仪器采用220V电源，请确认您的电源为稳定的220V。

如果电源不稳定的话，有可能对仪器造成严重损害！请确认您的电源严格接地，我们提供给您的电源线为三芯带地线电源。

请不要改动该三芯电源线结构，否则有可能对仪器造成严重伤害！3.安装：如果确认仪器包装无问题，且与合同相符，则可以进行安装。

请将仪器安装在一个干燥，水平，常温环境中。

尽量避免灰尘和化学药品对仪器的损害。

仪器与其他物品的距离不少于15厘米，以保证仪器冷却的需求。

将电源线等附件拆包装，待用，依据后续章节继续操作。

第二节技术规格1.外观尺寸：宽：17＂高：11＂长：＂2.重量：47磅3.电气规格：电源：220V，单相，7A耐熔保险交流：频率固定在1MHZ电压： 0 – 75 V （从零到峰值）脉冲时间：0 – 99 秒直流：高电压模式（HV）电压： 10 – 3000 V （峰值）脉冲时间：1 – 99 毫秒低电压模式（LV）电压： 10 – 500 V （峰值）脉冲时间：1 – 99 毫秒0.01–毫秒脉冲次数：1 – 994.前面板控制介绍：第三节操作细则1.注意事项：请严格按照下述操作细则对ECM 2001进行操作。

在没有连接打印机的情况下，严禁按“PRINT SETTINGS”按钮！否则仪器将会锁死。

如果打开仪器电源的时候，发现“MANUAL START”按钮处于亮灯状态，请马上关闭电源！2.程序或步骤（操作过程）高电压警告：为了实际操作和安全的原因，在手动或自动操作模式中，ECM 2001在施加交流电场或直流脉冲的过程中，请不要接触任何电缆或电极连接处，当需要连接或拆下电缆时，要检查系统不在操作中或处于备用状态。

BD Bioscience Discovery Labware BD Biocoat TM产品使用指南BD Biocoat TM Matrigel TM 基质BD Biocoat TM细胞相关检测系统目录1BD Biocoat TM Matrigel TM 基质1.1 Matrigel基底膜基质胶 (1)1.2 高浓度HC Matrigel基底膜基质胶 (3)1.3 人来源细胞外基质 (4)1.4 Ⅲ型人重组胶原蛋白 (5)1.5 高浓度层粘连蛋白/巢蛋白混合物 (6)1.6 人胚胎干细胞专用的Matrigel基质 (7)2BD Biocoat TM细胞检测相关系统2.1 Biocoat肿瘤侵袭系统 (8)2.2 Biocoat血管生成系统：内皮细胞侵袭 (10)2.3 Biocoat血管生成系统：内皮细胞迁移 (12)2.4 Biocoat血管生成系统：内皮血管形成 (14)Caco-2检测系统 (15)2.5 BiocoatHTS附录：细胞培养系统快速使用指南 (17)Biocoat TM Matrigel TM常见问题 (17)1BD Biocoat TM Matrigel TM 基质1.1Matrigel基底膜基质胶BD产品货号：354230 354234 356230 356231 356234 356235 356237储存和运输：-20度储存，干冰运输。

操作指南BD采用专利技术，从富含胞外基质蛋白的EHS小鼠肿瘤中分离出BD Matrigel 基底膜基质，其主要成分由层粘连蛋白，Ⅳ型胶原，巢蛋白，硫酸肝素糖蛋白等组成，还包含生长因子和基质金属蛋白酶等。

BD Matrigel基底膜基质在室温条件下，聚合形成具有生物学活性的三维基质，模拟体内细胞基底膜的结构、组成、物理特性和功能，有利于体外细胞的培养和分化，以及对细胞形态、生化功能、迁移、侵染和基因表达的研究。

BD Matrigel 基底膜基质形成的三维培养基质，可促进上皮细胞、肝细胞、Sertoli细胞、黑色素瘤细胞、血管内皮细胞、甲状腺细胞及毛囊细胞等的贴壁与分化。

多功能电穿孔系统1、工作条件：电源：100 – 120 VAC or 220 – 240 VAC, 50/60 Hz.功率：最大240W室温：0 - 35︒C,相对湿度：0 – 95%, (non-condensing)2、总则用于原核和真核细胞电转化3、详细技术指标及规格：3.1 系统配置：带有方波输出功能的主机、原核细胞组件及哺乳动物细胞组件，0.1/0.2/0.4cm电激转化杯。

该细胞转化仪用于原核细胞、真菌以及哺乳动物细胞的电击转化试验，包括外源DNA导入和细胞融合等。

技术要求和参数：*（1）系统输出波形：指数衰减和方波两种输出，适合不同应用需要。

（2）输出电压：10 – 3,000伏，最小调节量1伏。

*（3）电容容量：10 – 500伏：25 – 3275 μF以25 μF 递增，适合哺乳动物细胞需要。

500 – 3000伏：10, 25, 50 μF三种调节，适应原核细胞要求。

（4）电阻（并联）：50 – 1,000 Ω以50 Ω递增，及无限大设置*（5）样品电阻：在10 – 2,500 V时，最小20 Ω在2,500 – 3,000 V时，最小600 Ω（6）方波放电时间：10 – 500 V档位: 持续时间0.05 – 10 ms时，以0.05 ms递增，持续时间10 – 100 ms时，以1 ms 递增，可设1 – 10 次脉冲重复，0.1 – 10 sec 间隔500 – 3,000 V档位: 持续时间0.05 – 5 ms时，以0.05 ms递增，可设1 – 2 次脉冲重复，5 sec 最小间隔（7）主机：输出波形：指数衰减和方波两种输出波形输出电压; 200 – 3,000伏放电容量：10, 25, 50 μF，三档调节样品电阻：最小20 Ω，在10 – 2,500 V档位，最小600 Ω，在2,500 – 3,000 V档位，方波放电时间：持续时间0.05 – 5 ms时，以0.05 ms递增，可设1 – 2 次脉冲重复，5 sec 最小间隔*实验方法预存：24种程序预设，方便用户优化自己的实验条件用户自定义方法储存：最多144个程序方便用户储存自己的实验方法。