ECM及Matrigel使用手册
Transwell侵袭实验
Transwell侵袭实验1.实验前一天将ECM胶（8-12 mg/ml）（Sigma）放于4℃冰箱中融化过夜2.实验时将所用的枪头在冰上预冷半个小时，ECM胶用预冷的无血清1640按1：9稀释（稀释至1mg/ml），每孔中加入稀释好的ECM胶40μl，放入37℃培养箱中，孵育5h3.水化基底膜：吸出小室中残余液体，每孔加入70ul含无血清1640培养液，37℃，30min，吸去培养基4.对数生长期的细胞，胰酶消化细胞，0.1%血清1640悬浮，计数，稀释至密度为1 106/ml，每空中加200μl细胞悬液5.24孔板下室一般加入600µl含30%新生牛血清的1640培养基6.24h后，弃去孔中培液，PBS洗2遍，甲醇固定20分钟，0.1%结晶紫染色15-20 min，用清水洗3遍以上，用棉签轻轻擦掉上层未迁移细胞7.400倍显微镜下随即五个视野观察细胞，记数个人觉得做Transwell就像小马过河一样，要讲究个体化，不同的细胞侵袭能力不同，胶的稀释倍数、细胞上样量、上下室血清浓度差以及孵育时间也不同。
对于你的问题，我觉的不管是在温箱中放5个小时或者是在超静工作台中风干，其主要目的是为了让Matrigel从液体状变成胶冻状，在温箱中放置5个小时后，拿出来上室中肯定会有液体残留，此时一定要将残留的液体洗干净，然后在水化一下基底膜。
(1) 吸去培养液，用棉签轻轻擦除上室聚碳酸酯膜内侧面贴壁细胞，用0.01MPBS 漂洗两次，4%多聚甲醛固定10 分钟；(2) 吸去固定液，将膜风干，每孔加0.6ml 0.1%结晶紫染液，室温中放置20 分钟；(3) 轻轻甩去染色液，用蒸馏水洗涤各孔，将上室取出，从聚碳酸酯膜内侧面用吸水纸吸干水分，自然干燥；(4) 测定前，将各实验组上室置入新的24 孔板中，每孔加0.6ml 33％醋酸脱色，充分振荡，每组设定5 复孔；同时设置调零孔（33％醋酸）；比色：每孔取脱色液150μl 加入96 孔板中，在570nm 处测定OD 值。
GE E-CELL模块用户手册
规 范
3. 规范
3.1 进水要求
单位 ppm ppmCaCO 3 ppm ppm ppm ppm ppm 进水要求 * 使用 E-Calc 计算 < 1.0 ** < 1.0 @ 最小产水量 ~ 标准产水量 < 0.766 @ 最大产水流量 < 0.5 < 0.05 < 0.01 <1.0 未检出 <1.0 未检出 <5
7.
8.
停机程序 ……………………………………………………………………………………… 15 8.1 E-CELL™ 模块在自动操作模式下停机 …………………™ 模块长期停机 ……………………………………………………………… 15 16 16 16 16 17 17 19 20 20 22 22 22 23 25 25 26 27
极水流 GEWPT 淡水产水 整流器
一小部分淡水供给水流经每个E-CELL™ 系统E-CELL™ 模块极水室，并 且在离开系统后将直接排放的水 GE水处理及工艺过程处理 由E-CELL™系统生产的高纯水 直流电源
人身安全防护措施
2. 人身安全防护措施
在E-CELL™ 模块及本使用手册中可以看到以下象形图
TEA - MK-3Pharm
TEA - MK-3 Mini
** 阅读该部分操作说明书 - 浓水的流量取决于回收率与进水硬度。 注意：E-Cell™模块的进水必须是 RO 产水或同等水质的水。
3.2
产水要求
倘若保证进水满足进水水质要求及按照操作说明对系统进行操作， E-Cell™模块可以生产 16MOhm.cm 的产水 （MK-3, MK-3 Mini） 或者 10 MOhm.cm 的产水 （MK-3 Pharm） 。
细胞电融合仪ECM安装操作手册
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。
Med-E-Lert Automatic Pill Dispenser 用户手册说明书
ENGLISH Med-E-Lert™ Locking Automatic Pill Dispenser Easy Set-up GuideMODELS1.0PREMIUM / 1.0PREMIUM-C / 1.0PREMIUM-W01WHAT’S IN THE BOX?02INTENDED USE03SET-UP INSTRUCTIONS 04GENERAL INFORMATION 05QUICK Q & A06TESTING FUNCTIONALITY 07WARRANTY INFORMATION 08ATTENTION09REPLACEMENT PARTSClear and/or Solid White Lids designed to assist you with the organization of your medications, along with reminding you when to take them. This pill dispenser is designed to assist with medication adherence, offering you and your loved one additional peace of mind.01WHAT’S IN THE BOX?02INTENDED USE03SET-UP INSTRUCTIONSyou to lock and unlock the device, allowing access to the pill tray and controls.12/24-Hour FormatsThe Med-E-Lert™ defaults to a24-hour format, also commonlyreferred to as military time. The“AM” or “PM” symbol will appear to the right of the time when the clock is in 12-hour format.Press and hold buttons 2 and 3simultaneously to switch between formats.1. Press and hold down button 1 until the hour »Scroll down minutes quickly: press and hold down button2.1. Press and quickly release button 1 and four»Scroll up hours quickly: press and hold down button 3.2. Once the hour is selected, press button 1again to finalize and move to set the minutes.»Go down by one minute: press and quicklyrelease button 2.»Scroll down minutes quickly: press and holddown button 2.»Go up by one minute: press and quicklyrelease button 3.»Scroll up minutes quickly: press and holddown button 3.3. Once the hours and minutes have beenselected for scheduled event(s), press andquickly release button 1 to move the nextalarm.4. Repeat steps one and two through theremaining five alarms if necessary.»If you do not need additional alarms, pressand quickly release button 1 through eachguide you where to place yourmedication(s).(Tip: group your medications/supplements by the time of day that you take them; this will assist you when it comes time to place them into the pill tray.)1x/day ring2x/day ringLow Battery Alert and Standby Mode When the batteries are lower than 4.0V, both the red light and low battery icon that appears at the top right of the screen will begin to blink, alerting you to change the batteries.»(Note: this icon will only display once it istime to replace your batteries; all four AAbatteries must be replaced with new onesand oriented in the correct direction.)»If there is no operation performed for oneminute, the Med-E-Lert™ will go intostandby/sleep mode to save battery life.To wake up your unit before a scheduledevent, either pick it up and tilt it as if youwere dispensing medication or press andquickly release button 3 on the interface.04GENERAL INFORMATION05QUICK Q & A06TESTING FUNCTIONALITYThe purpose of the test mode is to determineif your pill dispenser is functioning properly by testing its buttons, voltage, icons/display, volume, sound quality, motion switch, light, circuit board, and tray alignment. (Note: if you have alarms set, you will not be able to access the test mode; before you can access this mode, you will need to remove all alarms before this becomes accessible, and then reprogram them after you are done.)Testing Buttons1.Press and hold down button 2 for severalseconds until you see the colon blink rapidlyand “C1 0” displays on the screen.2.Press button 1 three times. This will count tothree.3.Press button 2 three times. This will count tosix.4.Press button 3 three times. This will count tonine.Testing Battery Voltage1.Once you see “C2” on the screen, a secondset of numbers will indicate the voltage. Forexample, “C2 53” indicates 5.3v.Testing Screen and Icons1.Once you see “C3” on the screen, all iconswill light up simultaneously and after acouple seconds will move to the next testautomatically.Testing Audible Alarm, Visual Alarm, and Rotation1.Once you see “C4” on the screen, the tray willrotate one compartment to the left, triggering the first audible and visual alarms.2.Turn off the alarm by tilting the unit as if youwere dispensing medications into your hand.3.Turn it back upright and the audible and visualalarms will repeat one more time.Testing Circuit Board1. Once you see “C5” on the screen, the number one will appear on the display. Do not press anything during this time. Once done, it will automatically move onto testing the next internal component.Testing Motion Switch Alignment1. Once you see “C6” on the screen, the device will display random numbers on the screen.2. The pill tray will rotate 10 compartments, testing the motion switch and aligning gears for functionality.4. Once again, turn off the alarm by tilting the unit as if you were dispensing medications into your hand.07WARRANTY INFORMATIONThe Med-E-Lert™ comes with a one yearlimited warranty. Please note that the following circumstances may void your warranty; warranty is non-transferable; unauthorized resale of this product is prohibited and voids warranty.»Failure to insert and orient batteries correctlyand using all new, matching batteries mayvoid warranty.»Forcing rotation of the tray may harm the unitand void warranty.»Immersing unit in liquid or placing it in adishwasher may void warranty.»Using chemicals or solvents to clean the unitmay void warranty.»Any abuse of the unit, including dropdamage, may void the warranty.»Keeping the unit in or around a heat source/microwave may void warranty.»Keeping unit in direct sunlight or underultraviolet light may void warranty.»Placing liquid or powder substances intocompartments may void warranty.08ATTENTION»Disclaimer: this product is not intended for use in the diagnosis of disease or otherconditions, in the cure, mitigation, treatment,or prevention of disease.»After five minutes of nonuse, the Med-E-Lert™ will go into standby mode and shut down the display. Press button 3 or tip the unit to wake up the machine. To put the machine back into sleep mode to extend battery life, simply press button 2.»If the device is left upside down for one minute, the unit will chirp until the Med-E-Lert™ is turned back to its normal position.If it remains upside down and it is time for a medication event to occur, the pill tray will not rotate until it is returned to its normal state.»Using this product for anything other than its intended purpose may void warranty.09REPLACEMENT PARTS**********************+1 (801) 285-80119265 South Highland Dr., #900250, Sandy,UT 84093, United StatesENGLISH。
E-Prime的使用简明教程
E-Prime的使用简明教程一、简介E-Prime有许多功能，包括编写程式(E-Studio)、执行程式(E-Run)、整理data(E-DataAid)、修复data(E-Recovery)、合并data(E-Merge)。
一般来说，要编写程式我们会使用E-Studio。
在程式集中执行E-Studio后会出现选单，除了可以选第一项开启一个新的空白实验或是选第二项使用范例模式一步一步建立实验程式，也可以选第三项开启一个已存在的实验程式来进行编辑。
二、基本操作介面通常我们选择开启一个新的空白实验后，会出现如下图之画面。
1.工具箱(ToolBox) :在下图最左边的蓝色区域就是工具箱，工具箱里的物件（包括图片、声音、文字、反应回馈…等）都可以依照实验的设计和需求，自行加入到实验之中。
2.结构视窗(Structure) :结构视窗中会以树状图的形式，显示出使用者在实验结构中放入了哪些工具箱中的物件。
实验执行时，程式会依据此视窗中(由上而下)之顺序，先后呈现出使用者所放置的物件。
3.属性视窗(Properties) : 在结构视窗下方的属性视窗，可显示在结构视窗中被使用者所点选（用滑鼠左键在物件上点一下）之物件的属性。
我们可以藉由属性视窗来更改结构视窗中物件的名称、呈现时间长短及位置、资料收集方式…等基本属性。
4.工作区 :下图画面中右侧的区域为工作区。
当我们用滑鼠左键在已放置于结构视窗中之物件上点两下时，则此物件就会呈现在工作区之中，此时也可以对物件的属性进行编辑。
三、编辑实验程式举例说明假设我们现在的实验，想请受试者先看一个中文字中的部件（如：言），再看一个中文字（如：试），并请受试者判断先前看到的部件，是否有出现在后来看到的中文字中。
因此，本实验的呈现顺序为:指导语画面→练习开始画面→练习(5个trial)→练习结束画面→正式实验(10个trial)→实验结束画面练习trial为: 哔声→部件→遮蔽（mask）→中文字→回馈→全黑画面正式实验trial为:哔声→部件→遮蔽（mask）→中文字→全黑画面开始编辑1.时间轴:首先，在SessionProc图示上按滑鼠左键一下，会出现属性视窗。
基质胶说明书
Discovery Labware, Inc ., Two Oak Park, Bedford, MA 01730, Tel: 1.978.442.2200 (U.S.)CLSTechServ@ /lifesciencesFor Research Use Only. Not for use in diagnostic or therapeutic procedures.GUIDELINES FOR USEPRODUCT: Corning ® Matrigel ® Basement Membrane Matrix High Concentration, 10 ml vial CATALOG NUMBER: 354248BACKGROUND: Basement membranes are thin extracellular matrices underlying cells in vivo . CorningMatrigel Matrix High Concentration (HC) is a solubilized basement membranepreparation extracted from the Engelbreth-Holm-Swarm (EHS) mouse sarcoma, atumor rich in extracellular matrix proteins. Its major component is laminin, followedby collagen IV, heparan sulfate proteoglycans, entactin/nidogen.1,2 Corning MatrigelMatrix HC also contains TGF-beta, epidermal growth factor, insulin-like growthfactor, fibroblast growth factor, tissue plasminogen activator,3,4 and other growthfactors which occur naturally in the EHS tumor. Corning Matrigel Matrix HC iseffective for the attachment and differentiation of both normal and transformedanchorage dependent epithelioid and other cell types. These include neurons,5,6hepatocytes,7 Sertoli cells,8,9 chick lens,10 and vascular endothelial cells.11 CorningMatrigel Matrix HC will influence gene expression in adult rat hepatocytes 12,13 as wellas three dimensional culture in mouse 14-17 and human 18,19 mammary epithelial cells. Itis the basis for several types of tumor cell invasion assays,20,21 will support in vivoperipheral nerve regeneration,22-24 and provides the substrate necessary for the studyof angiogenesis both in vitro 25,26 and in vivo .27-29 Corning Matrigel Matrix HC alsosupports in vivo propagation of human tumors in immunosupressed mice.30-32 Forfurther information, go to our website at /lifesciences.SOURCE:Engelbreth-Holm-Swarm (EHS) Mouse Tumor FORMULATION:Dulbecco's Modified Eagle's Medium with 50 g/ml gentamycinCorning Matrigel Matrix HC is compatible with all culture media STABILITY:Stable for a minimum of three months from day of shipment when stored at -20 CKEEP FROZENRECONSTITUTIONAND USE: Color variations may occur in frozen or thawed vials of Corning Matrigel Matrix HC,ranging from straw yellow to dark red due to the interaction of carbon dioxide with thebicarbonate buffer and phenol red. Variation in color is normal, does not affect productefficacy, and will disappear upon equilibration with 5% CO 2.Once Corning Matrigel Matrix HC is thawed, swirl vial to be sure that material isevenly dispersed. Handle using sterile technique. Place thawed vial of Corning MatrigelMatrix HC in sterile area, spray top of vial with 70% ETOH and air dry. CorningMatrigel Matrix HC may be gently pipetted using a pre-cooled pipette to ensurehomogeneity.Corning Matrigel Matrix HC may be used as a thin gel layer (0.5mm), with cellsplated on top. Cells may also be cultured inside the Corning Matrigel Matrix HC,using a 1 mm layer. Extensive dilution will result in a thin, non-gelled protein layer.This may be useful for cell attachment, but may not be as effective in differentiationstudies. Corning Matrigel Matrix HC can be used to assess in vivo angiogenic activityof different compounds by subcutaneous injection into mice (Corning Matrigel PlugAssay).2,8,25 The high proteinDiscovery Labware, Inc ., Two Oak Park, Bedford, MA 01730, Tel: 1.978.442.2200 (U.S.)CLSTechServ@ /lifesciencesFor Research Use Only. Not for use in diagnostic or therapeutic procedures.concentration augments the growth of tumors and also allows the Corning ® Matrigel ®Plug to maintain its integrity after injection. This keeps the injected tumor and/or angiogenic compounds localized for in situ analysis and/or future excision.Dispense remaining material into appropriate aliquots, using pre-cooled tubes, andrefreeze immediately. Avoid multiple freeze thaws. DO NOT STORE IN FROST-FREE FREEZER.CAUTION :Corning Matrigel Matrix HC will gel rapidly at 22o C to 35o C. Thaw overnight at 4o C on ice (Matrigel may gel at slightly elevated temperatures in a refrigerator). Keep product on ice before use, and use pre-cooled pipettes, tips, and tubes when preparing Corning Matrigel Matrix HC for use. Gelled Corning Matrigel Matrix HC may be re-liquified if placed at 4°C on ice for 24-48 hours.INJECTION PROTOCOL:1. It is critical to keep the Corning Matrigel Matrix HC and the Corning Matrigel/Cell suspension as cold aspossible, without freezing, prior to injecting into the mice. It is very important to keep the Corning Matrigel and the Corning Matrigel/Cell suspension as asceptic as possible throughout the procedure.2. For each recipient mouse, mix cells (2 x 105 or greater) and Corning Matrigel Matrix HC together in a finalvolume of 0.5 ml on ice.3. The cells should be in as small a volume as possible. Typically, 250 l ice cold medium containing 2 x 106cells/ml is mixed with 250 l ice cold Corning Matrigel Matrix HC.4. Inject the cells subcutaneously in athymic mice using a 19G needle for tissue samples and a 23G needle forcultured cells. The injections should be done quickly to prevent the Matrigel from solidifying.5. Rotate the syringe when withdrawing to prevent leakage. The needles will need to be changed frequently dueto blockage.NOTE: For more details on this application go to /lifesciences to access CLS-DL-CC-036 (Technical Bulletin 455: Methods for Implantation of Corning Matrigel Matrix into Mice and Tissue Fixation).CELL RECOVERY:Dispase (Catalog No. 354235), Corning Cell Recovery Solution (Catalog No. 354253)Most efficient recovery of cells growing on Corning Matrigel Matrix HC is accomplished using Corning CellRecovery Solution that depolymerizes the Matrigel Matrix within 7 hours on ice or with Dispase, a metalloenzyme which gently releases the cells allowing for continuous culture.REFERENCES:1.Kleinman, H.K., et al., Isolation and characterization of type IV procollagen, laminin, and heparan sulfate proteoglycan from the EHS sarcoma, Biochemistry, 21:6188 (1982).2. Kleinman, H.K., et al., Basement membrane complexes with biological activity, Biochemistry, 25:312 (1986).3. Vukicevic, S., et al., Identification of multiple active growth factors in basement membrane Matrigel suggests caution in interpretation ofcellular activity related to extracellular activity related to extracellular matrix components, Experimental Cell Research, 202:1 (1992).4. McGuire, P.G. and Seeds, N.W., The interaction of plasminogen activator with a reconstituted basement membrane matrix and extracellularmacromolecules produced by cultured epithelial cells, J. Cell. Biochem., 40:215 (1989).Discovery Labware, Inc ., Two Oak Park, Bedford, MA 01730, Tel: 1.978.442.2200 (U.S.)CLSTechServ@ /lifesciencesFor Research Use Only. Not for use in diagnostic or therapeutic procedures.5.Biederer, T. and Scheiffele, P., Mixed-culture assays for analyzing neuronal synapse formation, Nature Protocols, 2(3):670 (2007). 6. Li, Y., et al., Essential Role of TRPC channels in the guidance of nerve growth cones by brain-derived neurotrophic factor, Nature, 434:894(2005).7. Bi, Y., et al., Use of cryopreserved human hepatocytes in sandwich culture to measure hepatobiliary transport, Drug Metabo. and Dispos.,34(9):1658 (2006).8. Hadley, M.A., et al., Extracellular matrix regulates sertoli cell differentiation, testicular cord formation, and germ cell development in vitro, J.Cell Biol., 101:1511 (1985).9. Yu, X., et al., Essential role of extracellular matrix (ECM) overlay in establishing the functional integrity of primary neonatal rat sertolicell/gonocyte co-cultures: An improved in vitro model for assessment of male reproductive toxicity, Toxilogical Sciences, 84(2):378 (2005).10. Ireland, M.E., Quantification and regulation of mRNAs encoding beaded filament proteins in the chick lens, 16(8):838 (1997).11. McGuire, P.G., and Orkin, R.W., A simple procedure to culture and passage endothelial cells from large vessels of small animals,Biotechniques, 5(6):456 (1987).12. Bissel, D.M., et al., Support of cultured hepatocytes by a laminin-rich gel. Evidence for a functionally significant subendothelial matrix innormal rat liver, J. Clinical Invest., 79:801 (1987).13. Page, J.L., et al., Gene expression profiling of extracellular matrix as an effector of human hepatocyte phenotype in primary cell culture,Toxilogical Sciences, 97(2):384 (2007).14 Li, M.L., et al., Influence of a reconstituted basement membrane and its components on casein gene expression and secretion in mousemammary epithelial cells, Proc. Nat. Acad. Sci. USA, 84:136 (1987).15 Barcellof, M.H., et al., Functional differentiation and aveolar morphogenesis of primary mammary cultures on reconstituted basementmembrane, Development, 105:223 (1989).16. Roskelley, C.D., et al., Extracellular matrix-dependent tissue-specific gene expression in mammary epithelial cells requires both physical andbiochemical signal transduction, Proc. Nat. Acad. Sci. USA, 91(26):12378 (1994).17. Xu, R., et al., Extracellular matrix-regulated gene expression requires cooperation of SWI/SNF and transcription factors, J. Biol. Chem.,282(20):14992 (2007).18. Debnath, J., et al., Morphogenesis and oncogenesis of MCF-10A mammary epithelial acini grown in three-dimensional basement membranecultures, Methods, 30(3):256 (2003).19. Muthuswamy, S.K., et al., ErbB2, but not ErbB1, reinitiates proliferation and induces luminal repopulation in epithelial acini, Nat. Cell Biol.,3(9):785 (2001).20. Terranova, V.P., et al., Use of a reconstituted basement membrane to measure cell invasiveness and select for highly invasive tumor cells, Proc.Nat. Acad. Sci. USA, 83:465 (1986).21. Albini, A., et al., A rapid in vitro assay for quantitating the invasive potential of tumor cells, Cancer Research, 47:3239 (1987).22. Madison, R., et al., Increased rate of peripheral nerve regeneration using bioresorbable nerve guides and laminin containing gel, Exp. Neurology, 88:767 (1985).23. Xu, X.M., et al., Axonal regeneration into Schwann cell-seeded guidance channels grafted into transected adult rat spinal cord, J. Comp. Neurol., 351(1):145 (1994).24. Fouad, K., et al., Combining schwann cell bridges and olfactory-ensheathing glia grafts with chondroitinase promotes locomotor recovery aftercomplete transection of the spinal cord, The Journal of Neuroscience, 25(5):1169 (2005).25. Kubota, Y., et al., Role of laminin and basement membrane in the morphological differentiation of human endothelial cells into capillary-likestructures, J. Cell Biol., 107:1589 (1988).26. Maeshima, Y., et al., Identification of the anti-angiogenic site within vascular basement membrane-derived Tumstatin, J. Biol. Chem.,276(18):15240 (2001).27. Passaniti, A., et al., A simple, quantitative method for assessing angiogenesis and anti-angiogenic agents using reconstituted basementmembrane, heparin, and fibroblast growth factor , Lab Invest., 67:519 (1992).28. Isaji, M., et al., Tranilast inhibits the proliferation, chemotaxis and tube formation of human microvascular endothelial cells in vitro andangiogenesis in vivo, British Journal of Pharmacology, 122:1061 (1997).29. Kisucka, J., et al., Platelets and platelet adhesion support angiogenesis while preventing excessive hemorrhage, Proc. Nat. Acad. Sci. USA,103(4):855 (2006).30. Albini, A., et al., Matrigel promotes retinoblastoma cell growth in vitro and in vivo, Int. J. Cancer, 52(2):234 (1992).31. Yue, W., et al., MCF-7 human breast carcinomas in nude mice as a model for evaluating aromatase inhibitors, J. Steroid Biochem. Molec. Biol.,44(4-6):671 (1993).32. Angelucci, A., et al., Suppression of EGF-R signaling reduces the incidence of prostate cancer metastasis in nude mice, Endocrine-RelatedCancer, 13(1):197 (2006).CALIFORNIA PROPOSITION 65 NOTICEWARNING: This product contains a chemical known to the state of California to cause cancer.Component: Chloroform。
细胞电融合仪 ECM2001 安装操作手册
细胞电融合仪ECM2001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：ECM2001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：17.5＂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：0–75V（从零到峰值）脉冲时间：0–99秒直流：高电压模式（HV）电压：10–3000V（峰值）脉冲时间：1–99毫秒低电压模式（LV）电压：10–500V（峰值）脉冲时间：1–99毫秒0.01–0.99毫秒脉冲次数：1–99第三节操作细则1.注意事项：请严格按照下述操作细则对ECM2001进行操作。
在没有连接打印机的情况下，严禁按“PRINTSETTINGS”按钮！否则仪器将会锁死。
如果打开仪器电源的时候，发现“MANUALSTART”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，ECM2001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。
夏米尔操作指南4
夏米尔操作指南4夏米尔ROBOFIL-XXX型操作指南状态信号行:GEO 路径转换激活时(MIR-ROT-INV)不亮，无功能激活时橙色 ALV 电极丝垂直校准时绿色ALP 电极丝工件校准时绿色GEN 脉冲电源接通时橙色CTC 电极丝短路时绿色ALM 数控在报警时红色UrG 紧急停止按钮按下时红色EXE 执行程序时黄色WIR 启动走丝时黄色显示当前使用的检查方式和时间MDI 键盘输入MEM 存储器JOG 手动Ready 准备就绪Start 执行Hold 中断用户参数SCF 缩放系数度 1 ROT 旋转角 0 MirX 对X轴镜像 0 MirY 对Y轴镜像 0 InvXY XY轴交换 0 TFE 考虑偏移 1 TER 考虑斜度角 1 ATH 自动重穿线ART 自动重启动SIM 空运行MLK 无移动模拟(程序校验)BLK 单段执行BLD 跳段有效，表示程序前有/的就不执行(如/M00) OSP 考虑可选停止制作:尹承效夏米尔ROBOFIL-XXX型操作指南BLK 打勾后，ISO程序单段执行OSP M01无效打勾后，M01有效(M01为暂停)BLD /开头的语句执行打勾后，/开头的语句跳过0、自动穿丝无效即断丝后不会自动穿丝ATH 1、断丝后回起割点自动穿丝再空走到断丝点切2、如果穿丝穿不进就直接加工下一个孔VSIM 空运行速度(6-900)1、机床坐标操做指令:SMA，X10 设制X轴机床坐标值为本10.00 SMA 设定机床坐标值X Y U V 同时为零MOV，X10 机床坐标系中的绝对移动将机床坐标X移到10处(注意无插补，各轴速度一样，路徑不是线性的) MOV XYUV都回零MOV，Z10 Z轴机床坐标移到10处绝对 Z轴不能同其他轴联动移动MVR 机床坐标的相对移动MVR，Z10 Z轴在现在的位置上向上移动10mm(不考虑转换、镜像、缩放，如果数控系统先前为绝对方式，移动后重新回到这一方式)制作:尹承效夏米尔ROBOFIL-XXX型操作指南CTR，X20 在此位置X轴向正方向加工20mm EDG，X-，X0.1252、工件坐标操作指令:SPA，X1 更新坐标SPA 同时更新XY的坐标为零MPA 绝对移动(不带值XY同时移动到零处) MPR 相对移动CPA 绝对加工CPR 相对加工3、预定义的移动GOH，Hh 按照工件高度H移动Z轴以定位喷嘴位 SEP，CPp 存储绝对坐标系中当前点的坐标p=点号，从0-5 GOP，p 移动到所定义的点位上，p=点号，从0-54、工艺和加工规准TEC<，表名> 激活含有准备使用的规准的工艺 WIR<，表名>激活准备使用的电极丝特性表HPA，h 修改当前高度h=新高度单位mm REX，Ee,(Hh)在所用工艺表中选择工艺规准Ee可选修改当高度(Hh) CLE(,c) 引入附加间隙(c=附加间隙，mm) CLE 设定附加间隙为零制作:尹承效夏米尔ROBOFIL-XXX型操作指南5、辅助功能AUX，m 辅助M功能(m=功能号)WTC 电极丝准备和切(拉)断WPR 断丝后的穿丝准备THR 自动穿丝(在WCT或WPR或人工丝准备之后) MDI模式常用指令:CT 相应的G\M代码SMA，XO，YO G74X0Y0 设定机床坐标的0，0点 SPA，XO，Y0 G92X0Y0 设定工件坐标的0，0点 MOV，X0，Y0 G75X0Y0 移动到机床坐标的0，0点 MVR，X2.0 X 方向相对移动2mm THD M60 穿丝WCT M50 剪丝SEP，CP1 G910 A1 设定当前点为1号点 GOP，1 G911 A1 从当前点移动到1号点EDG，Y，Y-0.125 Y方向对边，并把所对面设0CEN，R45 45度找孔中心 MID，R90 90度方向找中 CTR，X10 X方向相对切割10mm简单加工举例:TEC，LT25AREX，E2，H25制作:尹承效夏米尔ROBOFIL-XXX型操作指南CTR，X10程序执行:ZCL 加工计数器设回到零 SIM，I 激活(i=1) 不激活(i=0) EDG,s轴(，轴v) 找边CEN(,Xx)(,Yy)(,Rr) 找孔中心MID(，Xx)(,Yy)(,Rr) 在两个平行面之间找中GG00 快速移动G01 直线插补G02 顺时针圆弧插补G03 逆时针圆弧插补G04 暂停G09 准备停止一次移动到位 G10 指定偏移量/可选责程序跳步值 G11 自动加工顺序G17 编辑面选择G20 英制输入(英寸)G21 公制输入(mm)G22 存储行程极限功能ON(有效) G23 存储行程极限功能OFF G28 返回到第一参考点G29 从参考点返回到用户点 G30 返回到第二、第三、第四用户点 G32 存储当前点为用户第二、第三、第四参考点G33 设定用户点2，3，4G40 取消丝径补偿G41 电极丝左补偿G42 电极丝右补偿G46 最佳反向控制ONG47 最佳反向控制OFFG48 自动角部倒圆ONG49 自动角部倒圆OFFG50 取消斜度G51 左锥度G52 右锥度G53 在绝对坐标系中移动G61 恒定拐角半径制作:尹承效夏米尔ROBOFIL-XXX型操作指南G62 锥形拐角半径(最小) G63 锥形拐角半径(平均) G64 (最大) G65 宏调用(局部)G66 宏调用(全程)G67 取消宏调用(全程) G68 切割进给方式G69 准确停止方式G70 找边G71 找孔中心G72 找槽中心G73 电极丝校准和导向器设定循环 G74 设定测量点/机床坐标系设定 G75 在机床坐标系中定位 G76 定位-找边点类型1或2 G77 定位-测量点类型1、2或3 G78 定位-拐角点G79 计算工件倾斜角G86 取消支撑功能G87 “用支撑保留废芯”方式G88 “切除支撑”方式G90 绝对方式指令G91 相对G93 局部坐标系设定G94 恒速进给G95 伺服进给MM00 停止M01 可选停止(可用来建立支撑以保留废芯)M02 程序结束M06 穿丝水射流ON M07 上导电块退回ON M14 重穿丝块初始化 M15 锥度方式编成 M16 穿丝射水OFF M17 上导电块回退OFF M23 几何精度策略OFF M24 几何精度策略ON M27 保护策略OFF制作:尹承效夏米尔ROBOFIL-XXX型操作指南M28 一级粗加工策略ON M29 二级粗加工策略ON M30 程序结束和重绕 M31 预置计时器M32 检查水的离子度(电导率) M33 检查水的温度M34 工作液槽上水M35 放水M36 工作液槽位记忆+加载液位ON M37 加载液位OFF M40 空运行状态M42 走丝OFFM43 上、下冲液OFF M44 丝张力OFF M50 CUT 切丝M59 穿丝准备M60 穿丝M68 关闭循环泵M69 接通循环泵M70 轨迹返回功能M70 《执行有效》信号的管理 M71M73M74M80 加工状态M82 走丝开M83 下冲液开M87 存储功能1 ON M88 存储功能1 OFF M89 存储功能2 ON M90 存储功能2 OFF M91 遥控器报警有效 M92 遥控器报警无效 M93 遥控器报警接通ON M94 遥控器报警断开OFF M95 脉冲输出功能M96 完成加工M97 镜向反向加工M98 子程序调用M99 子程序结束制作:尹承效夏米尔ROBOFIL-XXX型操作指南穿丝射流水调整:EXE-service-calibration cyeles-wire calibration 激活卸下丝移动上加工头到相对于待加工工件所需的穿丝位置上接通穿丝射流水手动移动U/V轴以便使穿丝射流水对准下导向器的中心并在其周围分布均匀按memorize threading (记忆穿丝)ZUV 键，保存达到的位置存储UV位置;其值在穿丝操作中会自动用到。
modified matrigel plug assay 原理
modified matrigel plug assay 原理全文共四篇示例，供读者参考第一篇示例：Modified Matrigel plug assay是一种用于研究新药物对肿瘤生长和血管生成的影响的实验方法。
该方法是在原Matrigel plug assay 的基础上进行了改进和优化，能更准确地评估药物对肿瘤和肿瘤相关血管生成的抑制作用。
本文将从原Matrigel plug assay的原理入手，介绍Modified Matrigel plug assay的原理及其应用。
Matrigel plug assay，又称体内Matrigel植入实验，是一种利用Matrigel（一种基质蛋白）植入小鼠体内，观察其引起的血管生成和细胞浸润的实验方法。
Matrigel是来源于大鼠肿瘤细胞和实验室永生化细胞的培养基底物，富含细胞外基质蛋白，并且易于凝胶化。
在体内植入Matrigel后，它能够模拟肿瘤周边的细胞外基质环境，促进血管生成及肿瘤细胞的生长和浸润。
传统的Matrigel plug assay也存在一些局限性，如Matrigel固化时间不稳定、难以准确测量血管生成、难以控制植入物大小等。
为了克服这些局限性，Modified Matrigel plug assay被开发出来。
该方法将Matrigel与聚合物（如明胶、PEG）混合，形成一种更稳定的凝胶，并且可以通过调节加热时间和温度来控制凝胶化速度和密度。
还可以通过预先在Matrigel中注射肿瘤细胞或用抗血管生成药物处理Matrigel，以模拟不同的实验条件。
Modified Matrigel plug assay适用于评估抗肿瘤药物对血管生成的干扰作用，以及研究肿瘤微环境对肿瘤生长和转移的影响。
通过这种方法，研究人员可以更全面地了解药物对肿瘤相关血管生成的影响机制，为开发新的抗肿瘤药物提供重要参考。
该方法还可以用于评估患者肿瘤组织对不同药物的敏感性，为个体化治疗提供实验依据。
Kegel说明书
首先，让我们来测试你的敏感度模式一敏感度测试将帮助你规划练习项目。
使用探头，增加压力毫安，（+号按键，这是测试敏感度）如果压力低于50毫安时觉得痒就进入到模式二。
如果低于50毫安不能感觉到痒就可能已经存在神经损伤。
一天用模式一调至50毫安测试两次，连续测试四周，再进入模式二模式二高级敏感度测试。
如果在50毫安以下感到痒则进入凯格尔治疗项目，如果在50毫安以下不能感觉到痒则可能已经存在神经损伤。
一天用模式二50毫安测试两次，连续测试四周。
如果依然无法有任何痒的感觉，请咨询医生以获得建议。
建议采用什么锻炼项目？盆底肌锻炼模式三每天进行盆底肌锻炼一次或两次，开始增强你的盆底肌。
每天开启模式两次，连续六周，再进入下一模式模式四加强训练难度，使用探头获得最佳效果。
每天两次，如果条件允许，请坚持锻炼六周。
模式五每周保持采用模式三和四保持锻炼成效，每周只需要锻炼两次。
亲密测试模式六强化阴道壁和紧致度：每天使用探头一到两次。
如果成果满意，每周请坚持第五模式锻炼两次。
模式七性爱过程中让肌肉收紧更长时间：每天使用探头一到两次。
如果成果满意，每周请坚持用第五模式锻炼两次。
模式八增强敏感度：这是很有力的锻炼，请每天锻炼一次。
探头压力请勿超过50毫安或者让电击片超过15毫安。
将电击片设置在图标“后置四片”档位。
如果成果满意，每周请坚持用第五模式锻炼两次。
压力性尿失禁、遗尿、尿频模式十每天使用探头一到两次，连续使用六周，然后可以进入到下一模式。
模式十一每天使用探头两次，若条件允许，请连续使用六周，可以获得最佳锻炼效果。
采用模式十和模式十一各自锻炼连续六周后，如果效果满意，可以每周锻炼模式五两次以维持锻炼效果。
急迫性尿失禁、尿频或压力性尿失禁、遗尿模式十二每天使用皮肤探头一到两次，可以镇静膀胱。
图标保持在“4 back（4back elctrodes）”状态持续六周（如有必要，可延长时间），每周维持模式五锻炼模式两次以保持锻炼效果模式十三静坐、行走时与肌肉锻炼结合，请参考用户指南中的模式。
GrowlerWerks uKeg 128 Pressurized Growler 产品说明书
U K EG™Owner ’s Manual64 & 128Pressurized GrowlerW E L C O M E T O Y O U R U K E G2Hello from the founders of GrowlerWerks.We’d like to personally welcome you to your new uKeg. Thank you for making GrowlerWerks your choice inkeeping beverages fresh and carbonated for more than two weeks. The uKeg is a unique product that gives you the functionality of a full-bar system, letting you be your own bartender wherever you go. Vacuum insulated so your beverage stays cold all day, the uKeg is perfect for keeping craft beer fresh or holding your own cocktail creations and then taking it with you to share with friends.We hope you enjoy your uKeg for years to come and are here to support you along the way. The uKeg will require some maintenance over its lifetime, and some easy upkeep procedures will be required as components wear. The uKeg comes with a one-year warranty for manufacturing defects, but we will always support you with free email and phone support. Check out the how-to page on our website for a wealth of information on caring for your uKeg and to access our technical-support system.So please go forth and enjoy your uKeg. May your beer always be fresh, cold, and carbonated!Cheers,Brian, Evan, and ShawnIncluded with your uKeg • 2 CO • CO • Regulator cap • uKeg• Surgeon General’s warning (required in some states)• Owner’s manual • Felt protective bag • GrowlerWerks sticker • Spare cartridge sealInstall cartridge • Before heading out.• Ensure dial is in off Charge cap• Tighten sleeve quicklyFill uKeg• First, rinse withCap uKeg• Turn pressure up toPour and enjoy• Tilt glass, hold close to1.2.3. 4. 5.Do not use without CO2Do not store or transport uKegTap HandleTap BodyTap LockGooseneck Sight Guard Sight TubeUpper Fitting Pressure Gauge NutPressure Gauge Lower FittingPressure Dial Carry HandleVPR CapVariable Pressure Regulation CapCO 2*Cartridge Sleevether gases may be used:rgon can be used for non-carbonated beverages.O cream chargers won’t carbonate but can yield a smoother character.RISKS & WARNINGS se precaution when handling CO 2 cartridges and follow all instructions. Keep bottom of cartridge sleeve • R isk of cold burns – CO 2 gas and used cartridges can be very cold. Use caution when handling. Always • D o not expose to sun or heat.• D o not store above 49°C/120°F • K eep out of reach of children.Cap O-ring tight, smoothCartridge seal rounded, smooth, cleanis tight.to bottom out.counterclockwise.ridges point down.What to expect from your cap:• Cartridge will last for one growler fill, with some left over that can be used during cleaning (see page 7).Applying a food grade silicone grease to seals will extend their life.Pour a good beer• Open tap fully.• Let go of tap handle.it’s OK if it gets wet.that’s OK.Insulation• Stays cold 8 hoursor more.Empty bottle and rinseall parts.• Thoroughly rinse tap lock.Rinse cap and sleeve withhot water (< 120° F).6.2.Shake with tap open, facing downward.3.Allow to dry, and storewith cap off and tap open 7.Hand Wash OnlyFill with warm water under 120° F and 1 cleaning 4.Remove cap and rinse uKeg with hot water (< 120° F). • R epeat 3 times.Install cap, charge briefly, flow a little water through 1.5.* S ee page 14 for detailed uKeg diagrams.Stretch O-ring just enough for installation.Use a butter knifeor keys if you do not have a uKeg tool.Use 1.5mm allen wrench.stainless steel bottle in a thin layer and then covered with a protective lacquer finish.Your copper uKeg should not corrode or turn green, but it can be expected to darkenslightly over time.MAINTAINING YOUR UKEG Removing the Tap Handle910F O R C E C A R B O N A T I N GForce carbonating homebrew:1 F ill the uKeg to the fill line with your beer after fermentation.•Do not add priming sugar.•lf you bottle, add the priming sugar to the bottling bucket after filling the uKeg.•Remember to adjust your sugar addition for actual volume of beer you’ll be bottling.2 I nstall CO 2 into cap and install cap onto uKeg.3 Purge oxygen from uKegs:•Turn on cap to 1-2 psi then turn off again. •Loosen cap slightly to vent pressure. •Tighten and repeat once.•Tighten fully and turn up pressure.4 Turn up pressure to 15psi.5 Swirl gently then refrigerate.•Excessive shaking can allow beer into cap.6 S wirl frequently to accelerate carbonation.7 B eer will be fully carbonated in 2-3 days.7 R educe carbonation to desired level.•Indicated pressure will lower only after a little beeris poured.8 A fter first pour, uKeg may run out of pressure as all of the CO 2 has gone into the beer.•For the uKeg 64, you can purchase the 16gm sleeve and use a 16gm cartridge to avoid this.9 Open uKeg and install new CO 2 cartridge.10 Purge O 2.11 E njoy!Instructional video on our website: /howto11COCKTAIL RECIPESRed BrutIngredients:• 1 uKeg ™ 128• 1 Bottle – Wild Roots Raspberry Infused Vodka (or any vodka + raspberries)• 3 Liters – Torre Oria Brut (champagne, moscato, or prosecco)• 1 Lemon • 1 Box Raspberries• 1 Bundle Mint LeavesInstructions:Pour entire bottle of Wild Roots Raspberry Infused Vodka & 3 liters (four 750 ML bottles) Torre Oria Brut into uKeg 128.Charge the cap with CO 2 (using a 16-gram cartridge ensures you’ll have enough CO 2 to carbonate and dispense).Grab your flute.Drop 3 raspberries and a leaf of mint into flute (slap mint to charge flavor).Pour your beverage into flute, and garnish with lemon peel.Enjoy.WARNING: This combination is meant for those who enjoy the more tart side of life. The bite is as strong as its bark. To sweeten that, we suggest using a moscato, 5 tablespoons of honey, or both!Eastside MuleIngredients:• 1 uKeg ™ 64• 1 Bottle – Eastside Distilling Below Deck Ginger Rum • 4 Pack – 12 oz Reed’s Extra Ginger Beer• 1 Bottle Natural Lime Juice (found at any local grocery store)Instructions:Measure 20 ounces of Eastside Distilling Below Deck Ginger Rum, 40 ounces of Reed’s Extra Ginger Beer & 4 ounces of lime juice.Pour measurements into uKeg 64.Stir these ingredients for 30-40 seconds.Charge the cap with CO 2 (using an 8-gram cartridge ensures you’ll have enough CO 2 to carbonate and dispense).Place the cap on the uKeg and turn the pressure up to 15 psi, which will keep the correct level of carbonation in the ginger brew and ensure you taste that kick unique to a moscow mule.Grab a glass, drop in a few ice cubes, pour, sip, repeat.*You can adjust the dial to automatically maintain perfect carbonation.*Keep uKeg in refrigerator to ensure fresh pours to the last drop.TROUBLESHOOTING SOLUTIONPressure gauge only reads 0 psi or never changes.• Dial may be temporarily stuck. Tap on face of gauge while under pressure.• Beer may be dried in the gauge. Follow pressure gauge cleaning procedure.• Gauge may be damaged. Contact technical support for assistance.Pressure gauge stuck above 0 psi when off.• Beer may be dried in the gauge. Follow pressure gauge cleaning procedure.• Gauge may have been overpressurized or developed an internal defect. Contact technical support for assistance.• If damaged by a drop or impact, a new pressure gauge may be purchased on the website.Tap continues to drip without stopping when closed and locked.• A few drips after closing the tap are normal, but it should not drip over time.• Debris may be upsetting the internal tap seal. Follow tap cleaning procedure.• If uKeg continues to drip after cleaning, internal O-ring may have worn through use and be in need of replacement. Replacement seals can be purchased from the website.• If the above do not address the problem, please contact technical supportfor assistance.Beer pours foamy every time (more than ¼ cup of foam).• First pour may be foamy if the beer line is full of air after being filled or the tap assembly has been allowed to warm up. Second beer should pour fine, if that’s the case. Always fill uKeg with tap open and keep out of the sun.• Beer may be highly carbonated from bar or brewery. If beer in uKeg is pressurized at 5 psi but came from brewery beer lines pressurized at25 psi, beer will pour foamy until the pressure equalizes after a few days in the uKeg.• Be sure to open the tap handle fully and let go during the pour. Attempting to meter the flow during the pour can lead to foaming.• Beer line may be dirty or clogged. Clean the tap assembly.• If foaming only after the beer level drops below the upper fitting, there is an internal leak in the uKeg. Contact technical support for assistance./howtoSight tube assemblyTap HandleTap Handle Set ScrewRocker Nut Rocker Seal RockerLock Pin Tap BodyShuttleShuttle O-ring VPR Cap bottom assemblyPin SeatCap O-ringSize -224High Side Pin Pin Spring Piercer O-ringPiecerCartridge Seal Reatining NutTap AssemblyUpper Gooseneck SealLower Gooseneck Seal Sight Tube O-ring (x2)Lower Fitting O-ring Lower Fitting AssemblyRiser Tube O-ring (x2)Riser Tube Upper Fitting Sight Guard Riser Socket Sight Tube Gooseneck VPR Cap top assemblyCap Screw(x4)Cap BodyLow Side SpringSpring HatDial ScrewDialDrive ScrewCap GripDiaphragmDiaphragm O-ringSize -126Lower FittingPressure Gauge O-ring Pressure GaugePressure Gauge Retaining NutPressure gaugeassemblyWARRANTY & REGISTRATIONRegister your uKegRegistering your uKeg gains youaccess to customer service. It alsoensures that your one-yearwarranty is properly applied./registrationVisit for more helpful uKeg TMproducts.CO 2 cartridges Bar mats Maintenance kits Cleaning tablets Wooden carrying case Wooden tap handlesV e r s i o n 2.0。
ECM及Matrigel使用手册
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。
卡林燃料技术有限公司 90000型号微处理器操作多接触温度限制控制器说明书
Data sheet126 Bailey RoadNorth Haven, CT 06473Phone 203-680-9401Fax 203-680-9403Carlin Combustion Technology, Inc.T ech support ©Copyright 2016 — Carlin Combustion Technology, Inc.Specifications• Carlin’s Model 90000 microprocessor-operated, multiple-contact tem-perature limit controls are available in three configurations, described below. Each model provides two contacts — one for operating limit and one from high limit.• Refer to separate product listing sheets for pre-defined models, or re-quest a control to meet your specifications, within the available ranges listed below.Control modelControl power input Contacts Contact ratingMN90000B 071916Model 90000B, C and CE Microprocessor Temperature Controls — Data sheetPower, Off – (How to enter = No power is applied to the red-white wire)• All lights and screen will be off. Power, On – (How to enter = Power applied to red-white wire)• System will power on and enter NORMAL OPERATION MODE Self-test – When power is applied, the 90000 performs a self-test, checkingsensor(s) and microprocessor and verifying limit contacts are open. The power-up test lasts from 3 to 5 seconds. The 90000 continues diagnos-tic checking during the operating cycle as well. Any self-check failure causes a lockout (see below). Call for Heat – When the temperature at the operating sensor is at or belowsetpoint minus fixed differential, the control powers the limit relay (break-on-rise contact closes; make-on-rise contact open).Stand By – When operating sensor reaches setpoint temperature or above,temp control will turn off the relay (break-on rise contact opens; make-on-rise contact closes). Normal Operation (Mode) – (How to enter = Initial starting mode)• The EZ-Temp will display the User Set Temperature. Display – (How to enter = Pressing UP or DOWN key in NORMAL MODE)• I n DISPLAY MODE, the EZ-Temp will display current temperature read-ing for 5 seconds, then return to NORMAL OPERATION Setting – (How to enter = Hold up and down buttons for 3 seconds while in NORMAL OPERATION) • I n SETTING MODE, the user is able to adjust the operating limit (via setting the set temperature). Once in this mode, the display will begin flashing the current set temperature. The display will increase one degree per UP or DOWN button push or 10 degrees per second when the button is held. Diagnostic – (How to enter = Power up the system with the UP and DOWNbuttons depressed. Once buttons are released, the system will move into diagnostic mode) • D IAGNOSTIC MODE displays recorded error codes. When powered in this mode, the display will show the newest recorded error code. As the DOWN key is pressed, the system will display error codes fromnewest to the oldest. When there are no more codes to display, the EZ-Temp will show a “E _ _” to indicate end of error codes. If the user continues to push the down button, the system will go back to the newest error code and begin displaying the error codes again from newest to oldest. The only way to exit this mode is to power cycle the system. To clear error codes (while in diagnostic mode), push and hold reset button for 3 seconds. To clear error codes (while in diagnostic mode), push and hold RESET button for 3 seconds.High Limit Temp – Fixed temp where the control will shut down and lockout(model dependent).High Limit Test – (How to enter = In NORMAL OPERATION, hold RESET, UPand DOWN buttons for 3 seconds) • D uring this operation, EZ-Temp will display a flashing 180 (or your current high temp). Pressing the DOWN button adjusts the high limit temperature. The high temp will decrease one degree for each press of the DOWN button. If the DOWN button is not pressed for a period of 10 seconds, the unit will return to normal operation. When the high limit is reached, the unit goes into hard lockout and displays the error code. Holding the RESET for 3 seconds clears the lockout and returns the high limit to default (180°). If the lockout condition has driven the unit into latchup, the RESET must be held for 30 seconds. Soft Lockout – (How to Enter = When any temperature sensor goes out ofrange or the system has detected a high temp condition and the primary and safety relays are still operational) • W hen in this state, it will display the error code but will leave thelockout LED off. If the condition that caused the soft lockout clears, the system will return to normal operation. Lockout – (How to Enter = Numerous Error Codes)• T he EZ-Temp will enter this state when an unsafe condition has oc-curred and the user must intervene and put the system in a safe state. Lockout can be cleared by pressing the RESET button for 3 seconds. Latchup – (How to Enter = Occurs after 3 LOCKOUTs have been detected)• T he system will enter a safe mode and will not exit the mode without user intervention. You can exit latchup mode by depressing the RESET button for thirty seconds. By exiting this way, the system will reset the lockout count back to zero.Set Temp – The target temperature of the appliance.Operation ModesMN90000B 071916Model 90000B, C and CE Microprocessor Temperature Controls — Data sheetMounting – 90000 controls mount to any standard well. Mountthe 90000 directly to a well (new or existing) with hardware supplied in the separate well mounting kit. See below for dimensions. Well Kits – Wells for 90000 sensors are available in the sizes shownbelow. Well kits include sensor mounting hardware designed to hold sensor securely in position. Sensors – Sensors are available separately for 90000.ConfigurationsError CodesError CodeReason01Primary (K2) relay is welded on. Note: If the control is reset from error code 01, the red LED will flash every 5 seconds indicating the control must be replaced.02Primary & Safety relays (K1 and K2) on. The control cannot be reset from error code 02 and must be replaced.03Primary & Safety relays (K1 and K2) are off 05Simulated High Temp lockout06Temp is above set high temp but below thermistor high (250°F)16AC line frequency out of toleranceError CodeReason20Temp sensor 1 out of range failure 21Temp sensor 2 out of range failure 22High limit temp sensor out of range failure 23Temp sensor 1 pin is open 24Temp sensor 2 pin is open 25High limit temp sensor is open07, 10, 11, 12, 13, 14, 15Internal hardware safety failure – unit should be replaced. Contact customer service for additional informa-tion. 1-800-989-22753"43/8"43/4"Model 90000B, C and CE Microprocessor Temperature Controls — Data sheetMN90000B 071916。
matrigel成胶原理 解释说明以及概述
matrigel成胶原理解释说明以及概述1. 引言1.1 概述在细胞生物学和组织工程学领域，三维培养环境对于模拟体内条件以及深入研究细胞-基质相互作用至关重要。
而matrigel作为一种常用的基质支架材料，被广泛应用于细胞培养、肿瘤生物学研究、组织工程以及药物筛选等领域。
本文旨在探讨matrigel成胶的原理和机制，并分析其在三维结构形成中的重要影响因素。
1.2 文章结构文章将从以下几个方面进行阐述：首先，我们将介绍matrigel的组成和特性，以便读者对其有更全面的了解；其次，我们将详细探讨matrigel在细胞培养中的应用，并对其与细胞相互作用机制展开分析；接着，我们将解释说明matrigel 的成胶过程，并对形成三维结构的机制进行深入解析；最后，我们将着重探讨在matrigel成胶过程中温度、pH值调控以及不同细胞类型和密度对结果的影响；最后，我们将对matrigel的成胶机制和应用价值做总结，并展望其在研究领域未来的发展方向。
1.3 目的本文旨在全面解释和概述matrigel成胶的原理和机制，希望读者能够深入了解matrigel在细胞研究和组织工程中的重要应用，并为进一步探索matrigel的潜力和发展提供思路与启示。
2. matrigel成胶原理：2.1 matrigel的组成和特性：matrigel是一种生物基质，由培养小鼠肺上皮细胞所分泌的外基质蛋白构成。
它主要由胶原、纤维连接蛋白、腺苷酸酪氨酸激酶等多种蛋白质组成。
这些组分使得matrigel具有独特的生物活性和结构特点。
2.2 matrigel在细胞培养中的应用：matrigel广泛应用于三维细胞培养实验中，能够模拟体内细胞生长环境。
其天然的三维结构提供了支持和导向作用，有助于实现多种细胞类型在体外形成组织结构，并促进细胞增殖、迁移和分化等生物过程。
2.3 matrigel与细胞相互作用机制：matrigel通过与细胞表面的整合素分子相互作用，介导着细胞-ECM（外基质）之间的信号传递过程。
Transwell是检测细胞侵袭能力的检测方法
Transwell是检测细胞侵袭能力的检测方法我们所使用的是Chemicon公司的ECM554，基质胶已经包被好了，买来就可以用，很方便，而且我们算过，跟自己买胶来包被价格相差不大。
这种胶4度下很软，37度则变得比较坚硬，因此用前应先放在培养箱中10分钟作用，使胶完全凝固。
该试剂盒对穿过膜的细胞进行荧光染色，再用裂解液裂解细胞后检测荧光值。
我们在实际使用的时候对穿过膜的细胞采用1％结晶紫染色，镜下计数细胞数目。
结晶紫也可以用33%醋酸溶解，再用酶标仪570nm检测，同样可以反应穿过细胞数因为基质胶的厚度直接影响穿过膜的细胞数，因此建议有条件的话还是买这类已经铺好胶的试剂盒，比较可靠。
说明书该公司网上可以下载，对原理也有比较详细的阐述，可以去查一下。
我们做的时候，上室用0.5%BSA的培养液，下室用5%血清的培养液，下室血清的浓度可根据细胞类型的不同进行调整，如果细胞侵袭能力强，可适当降低血清浓度，否则可适当提高血清浓度。
需要注意的是：1。
上室内的细胞数目要适当，过多，穿过膜的细胞会过多过快，如果用计数法的话将难以计数；最少也要保证在收样的时候，上室内还要有细胞存在。
具体细胞密度需要自己摸索。
2。
细胞计数的准确性对结果影响很大，各组间最好不要分开计数。
尽量用同一密度的细胞悬液给予不同处理，保证各组细胞量一致3。
对细胞的处理不可影响细胞生存。
否则难以肯定是细胞量的改变还是侵袭能力改变侵袭小室其实不止是transwell的，其它像Boyden chamber、Thincert也挺好用的。
我用过Thincert，个人感觉不错，而且价格便宜，是grelner的，孔径8微米用于24孔板的一个才45块人民币，而且可以零买，不想transwell，总是一箱一箱的卖，实在是太贵了！侵袭实验要结合实验目的，不一定非要包被的。
下面奉上用Thincert做侵袭实验的步骤（摘自《A quantitative cell migration assay using ThinCert™ cell culture inserts》，大家如需要，可用google搜之）：1. Prepare cell cultures according to standard cell culture procedures2. Starve cells overnight in serum-free medium with 0.2 % BSA3. Harvest cells and wash them twice in PBS4. Resuspend cells in serum-free medium with 0.2 % BSA to an appropriate final cell concentration (e.g. 106/ml)5. Place 24 well ThinCertTM cell culture inserts in the wells of a CELLSTAR® 24 well cell culture plate6. Add 600 µl culture medium with or without chemoattractant to each well of the cell culture plate7. Add 200 µl cell suspension to each cell culture insert8. Incubate the cell culture plate for 3-24 h in a cell culture incubator at 37°C and 5 % CO29. Remove the cell culture medium from each well of the cell culture plate and replace it by 450 µl serum-free culture medium with 8 µM Calcein-AM10. Incubate for 45 min in a cell culture incubator at 37°C and 5 % CO211. Remove the culture medium from the cell culture inserts12. Transfer the cell culture inserts into a freshly prepared 24 well cell culture plate containing 500 µl prewarmed Trypsin-EDTA per well13. Incubate for 10 min in a cell culture incubator at 37°C and 5 % CO2, agitate the plate from time to time14. Discard the cell culture inserts and transfer 200 µl of the Trypsin-EDTA solution (now containing the migratory cells) from each well into a black flat bottom 96 well plate15. Read fluorescence in a fluorescence plate reader at an excitation wavelength of 485 nm and an emission wavelength of 520 nm从第9步开始计数侵袭细胞时，这份说明书用的是荧光标记，但这种方法成本高而且繁琐。
ECM说明书
设备状态监测系统ECM 使用说明书北京万博振通检测技术有限公司出品声明：本软件是我公司自主开发，享有知识产权。
未经授权不得擅自使用或拷贝。
2010-8-26目录1.软件安装 (3)2.软件启动与测点设置 (4)2.1登录 (4)2.2 主画面 (4)2.3设备、测点设置 (5)2.4基准谱绘制 (5)3.主菜单功能 (7)3.1系统设置 (7)3.2设置数采器 (7)3.3数据回收 (9)3.4趋势分析 (11)3.5专业分析 (11)3.6报告 (15)3.7视图 (15)3.8帮助 (15)4.数据库拷贝与软件更新 (15)5.设备故障诊断专家系统（FDES） (17)6.ECM版本历史 (27)1.软件安装用鼠标双击光盘中的ECM文件夹下的setup.exe文件，开始软件安装。
安装过程中出现窗口：按”NEXT”“YES”继续直到出现选择安装路径界面：安装默认路径是C:\Program Files\BVM\ECM。
用户也可按‘Browse’选择其它路径进行安装。
2.软件启动与测点设置2.1登录选择开始菜单>>所有程序>>设备状态监测系统>>设备状态监测系统，第一次使用者进入以管理员administrator进入软件（双击用户名输入框见下拉菜单）。
用户可以根据实际需要，添加或修改用户。
添加密码后再次进入软件将会出现登录窗口：2.2 主画面图1 主菜单2.3设备、测点设置1部分是设备树，分四个等级：“总厂”“分厂”“车间”“设备”。
选中任意文字再次单击可以更改名称，3部分和1部分是联动的，用来设置和显示被选中（总厂分厂车间或者设备）图片。
其中总厂级别是唯一的，在总厂级别上右键可以添加新分厂；在新分厂级别上右键可以添加新分厂、添加新车间、删除***（被选中的新分厂名称）；在新车间级别上右键可以添加新车间、添加新设备、删除***（被选中的新车间名称）；在新设备级别上右键可以添加新设备、删除***（被选中的新设备名称）2部分上边是添加和显示测点参数和报警值，下边显示的是测点的数据，当选中某一测点数据时4部分同步显示此数据的波形和频谱。
基质胶matrigel matrix操作注意事项
注意事项亲爱的用户：您好！鉴于基质胶的操作环境要求比较严格，为保证您的实验顺利，特此对以下几个方面进行温馨提示：1.收货时，首先确认还有干冰，matrigel matrix成固态，液面水平，如有异常请及时拍照取证并联系我们；2.整个matrigel matrix只能分装一次，在分装前，要先放在4℃冰箱（最好先把matrigelmatrix插在冰上，然后放入4℃冰箱）一段时间待之融化（切记不要放在冰箱门上或者是经常要敞开口取放试剂的冰箱），且分装用的枪头、EP管等都要提前-20℃预冷（因为基质胶在10℃以上即会发生成胶凝固导致基质胶报废），整个分装过程要在冰上进行，且以后每次实验时拿出本次用量的已经分装好的基质胶用，该次实验用不完的基质胶要扔掉，不可以留作下次使用；3.实验人员的手心不要接触基质胶，如：要手持装有matrigel matrix的EP管的上方，防止体温使基质胶成胶凝固；4.普通matrigel matrix的浓度是8-12mg/ml，高浓度的matrigel matrix的浓度是18-22mg/ml，具体浓度可通过货号查询；5.以24孔板为例，做肿瘤细胞侵袭实验时每孔加入matrigel matrix 20-30ug（一般配成200-300ug/ml的工作浓度，然后每孔加入100ul即可）。
此时，matrigel matrix是稀释到很低的倍数（有的实验需要低至30倍稀释，具体最适浓度需根据自己的实验进行优化）。
铺胶后置于37℃培养箱放置1小时，取出后如果上层有液态培养基，需小心弃去上层的液态培养基即可使用（此时matrigel matrix沉在底部形成一小薄层）；6.做内皮细胞的成管实验一般把高浓度的matrigel matrix（18-22mg/ml）稀释到10mg/ml，由于普通浓度的matrigel matrix（8-12mg/ml）的浓度接近10mg/ml，所以有时也可用作成管实验；7.应使用传代次数较少、状态良好的细胞进行实验；8.以上内容非常重要，请务必重视。
Matriarch 序列器操作指南说明书
RECORD A SEQUENCE:•Follow steps 1-7 under the keyboard illustration.PLAYBACK YOUR SEQUENCE:•Set the MODE switch to the SEQ position.•Press the PLAY button (located on the left-hand controller), then press and hold down any note to playback your sequence.•Press the HOLD button to allow the sequence to continue playing after your hand is lifted from the keyboard.•The Arp/Seq RATE knob adjusts the playback speed.NOTE: You may transpose your sequence by playing a new note.MATRIARCH SEQUENCERNOTE: This will arm Sequence 1.NOTE: During playback, this step will be silent.NOTE: Matriarch provides a total of 12 sequence locations, which are accessibleby using the four-position SEQUENCE knoband the OCT / BANK switch.ADD A REST 3. This is the second note of your sequence. ADD A NOTE 4. Finally, press the REST button, play a note, and then press the RATCHET button a few times. To listen to your recording, follow the steps under PLAYBACK YOUR SEQUENCE.ADD A REST + NOTE + RATCHET NOTE: Pressing the RATCHET button multiple times will add that number of Ratchets (up to 8) to a single step.Press the TIE button and play thesame note (from Step 4) again. ADD A TIE 5. ADD A TIE (LEGATO)Press and hold a new note. While holding that note, press another note.NOTE: During playback, the transition between these two notes will be smooth, without a break. This method of adding a tie between notes willonly work in 1-Voice monophonic mode. ARM THE SEQUENCER 1. To arm the Sequencer for recording, set the MODE switch to the REC position. PLAY A NOTE 2. This is the first note of your sequence. NOTE: With the VOICE MODE switch set to 2 or 4 (paraphonic mode), you can record up to four notes simultaneously on a single step.C#0D#0F#0G#0A#0C#1D#1F#1G#1C#2D#2F#2G#2A#2C#3D#3F#3G#3A#3A#1C0D0E0F0G0A0B0C1D1E1F1G1A1B1C2D2E2F2G2A2B2C3C4D3E3F3G3A3B36. 7.PATCH TOREAR PANEL PATCH EXPLORATIONNOTES:•MOD WHEEL adds audio rate filter FM.•Patch works well with paraphonic sequences.•GLIDE set at 8 o’clock position.PATCH EXPLORATIONNOTES:•Create and play a sequence.•MOD WHEEL set at 100%.•Play in stereo and loud!PATCH EXPLORATIONNOTES:•GLIDE set at 9 o’clock position.•MOD WHEEL set at 100%.•S/H controls synced Pitch / FM.2NOTES:•Set ARP or create and play a sequence. (Try a DMAJ 7 starting on D2).•Use OSCILLATOR 3 and NOISE knobs in mixer section for sudden passing winds and melodies.•Play in stereo.•Use ATTENUATOR 2 for clusters while turning OSCILLATOR 1 in mixer section.PATCH EXPLORATIONPATCH EXPLORATIONNOTES:•Try adjusting oscillator waveforms so that each oscillator has a different shape.•Adjust Arp/Seq RATE to your desired tempo.PATCH EXPLORATION3 NOTES:•MOD WHEEL up.•Set up an ARP and play. (Try A2/C2/D2/G3).•ATTENUATOR 3 brings volume of “secondsequence” up.•Raise OSCILLATOR 4 to bring in deep phasersync colors.•Play in stereo.PATCH EXPLORATIONNOTES:•Record a sequence with 2 notes per step.•MOD WHEEL up.PATCH EXPLORATIONNOTES:•MOD WHEEL controls Cutoff Modulation and OSC detune.•GLIDE set at 8.5 position.PATCH EXPLORATIONNOTES:•Set-up an ARP and listen to it sway and drip around.PATCH EXPLORATIONNOTES:•Round robin on with RESET. (See pg 68 ofuser’s manual).•Take your time dialing in OSC tuning to achieve harmonius ARP playback.PATCH EXPLORATIONNOTES:•Modulation RATE knob controls the modulation of the second LFO’s rate.•Adjust Arp/Seq RATE to your desired tempo.NOTES:•Tune OSCILLATORS 1-4 in unison.•Create a polyphonic sequence incorporating rests.•Play at slow tempo. Dial ATTENUATOR 1 to bring in “second sequence”. •Dial ATTENUATOR 3 to filter for desired filter animation.•Change SEQ direction for interesting variations.•Use ratchet and tie for variation.PATCH EXPLORATION123PATCH TOREAR PANEL PATCH EXPLORATIONNOTES:•MOD WHEEL at 100%.•GLIDE set at 8 o’clock position.312NOTES:•MOD WHEEL at 50%.•ATTENUATOR 1 and 3 control left and right voice mix blend.•When you allow last chord to decay, unplug VCA 1 IN for descending “Space Echos.”•Adjust PORT / GLIDE for more stereo drama.PATCH EXPLORATION。
Matrigel使用常见问题
Matrigel使用常见问题本期为大家带来Matrigel使用的常见问题，满满的干货，务必仔细阅读啦Matrigel使用常见问题-FAQ⚑ Matrigel是什么，里面包含哪些成分？Matrigel是来源于EHS小鼠肉瘤的基底膜基质，其中含有约60%层粘连蛋白、30% IV 胶原和8%的巢蛋白，还含有基底膜聚糖、TGF-ß、表皮生长因子、类胰岛素生长因子、组织纤溶酶原及其他生长因子。
⚑ Matrigel有哪些种类？康宁Matrigel主要有常规标准型（Standard）、高浓度（HC）、无酚红（Phenol red-free）、生长因子减量型（Growth factor reduced, GFR）和人胚胎干细胞专用（hESC-qualified）几个类别。
•标准型Matrigel：浓度范围大致为8-12mg/ml•高浓度Matrigel：浓度范围大致为18-22mg/ml•无酚红Matrigel：不含酚红，产品完全融化后颜色透明•生长因子减量型Matrigel：Matrigel里包含的生长因子经过减量处理•人胚胎干细胞专用Matrigel：经过对hESC培养的测试，可以用于人胚胎干细胞的培养⚑如何选择Matrigel？⚑ Matrigel到货需要注意什么？首先确认是否由干冰运输，到货时包装箱中干冰是否足量，查看Matrigel液面是否基本水平，以及瓶身侧壁、瓶口有无Matrigel融化的痕迹，以确保运输质量；若发现问题，请及时拍照记录，并联系经销商反馈。
⚑如何储存Matrigel？收到产品并确认到货情况无异常后，请将Matrigel储存于-20℃冰箱，长期保存可置于-80℃冰箱。
切勿将Matrigel储存于自动除霜的冰箱中。
在第一次融化后请分装保存，应避免Matrigel反复冻融。
⚑ Matrigel颜色为什么会有不一样？无酚红Matrigel没有颜色，完全融化后应是透明的；含酚红Matrigel颜色可以在明黄色至暗红色之间变化，这些颜色变化是正常现象。
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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。
同时，Matrigel还能影响乳腺上皮细胞的蛋白表达，支持外周神经的新生和牛输卵管上皮细胞的分化。
高浓度的Matrigel适用于研究体内血管生成和肿瘤细胞迁移及肿瘤模型的建立等。
产品特性Matrigel基质会有色差变化（淡黄色到深红色），是由于酚红和碳酸氢盐与CO2作用引起的，但是与5％CO2平衡后色差即会减少。
冻融后，轻轻摇晃试剂瓶使Matrigel分散均匀。
所有操作均需在无菌环境下进行，试剂瓶瓶盖可用70％乙醇擦拭，并自然干燥。
应使用预冷的移液器以保证Matrigel呈匀浆状。
细胞可在0.5mm厚度的Matrigel基质层表面生长，也可在1mm厚度的Matrigel三维基质内生长。
过度稀释的Matrigel会形成非胶质的蛋白层，可以用于细胞贴壁，但不能用于细胞的分化研究。
注意：可将冻融后的Matrigel分装在多个小管，所有分装均需用预冷的冻存管，迅速冷冻并保存，避免多次冻融。
Matrigel在22-35℃温度环境下快速成胶，因此溶解时在4℃冰上过夜冻融（4度时会随着温度的上升部分成胶）。
所有用品在使用前需置于冰浴，必须使用预冷的移液管、吸头及小管操作Matrigel。
成胶后的Matrigel可以在4℃24－48小时后重新呈液态。
推荐包被与成胶方法注意：为了保证Matrigel基质膜的成胶性能与稳定性，稀释浓度不应低于1：3，可用无血清培养基稀释，Matrigel成胶后立即使用。
薄胶成胶方法：1、冻融后，用预冷的移液枪头混匀Matrigel基质成匀浆状。
2、将需要使用的培养板置于冰上，加入浓度为50μL/cm2生长面积的Matrigel基质。
3、在37℃放置30分钟，即可使用。
厚胶成胶方法：1、冻融后，用预冷的移液枪头混匀Matrigel基质成匀浆状。
2、将需要使用的培养板置于冰浴，将培养的细胞与Matrigel基质混合，用移液枪头使其悬浮于基质中。
加入浓度为150－200μL/cm2生长面积的Matrigel基质。
3、在37℃放置30分钟，可成胶。
可以加入细胞培养的基质，也可使细胞直接生长在胶表面。
薄层包被方法：1、冻融后，用预冷的移液枪头混匀Matrigel基质成匀浆状。
2、根据使用需要，采用无血清培养基稀释Matrigel基质。
根据实验需要确定最佳包被浓度。
3、将稀释的Matrigel基质包被于所需的培养器皿中，包被量至少覆盖整个器皿的生长表面。
室温下孵育1小时。
4、去除未结合的Matrigel，用无血清培养基轻轻地冲洗。
用BD细胞回收剂（354253）或离散酶（354235）可降解Matrigel基质，冰浴7小时后回收得到细胞。
1.2高浓度HC Matrigel基底膜基质胶BD 产品货号：354248 354262储存和运输：-20度储存，干冰运输。
产品特性Matrigel基质会有色差变化（淡黄色到深红色），是由于酚红和碳酸氢盐与CO2作用引起的，但是与5％CO2平衡后色差即会减少。
冻融后，轻轻摇晃试剂瓶使Matrigel分散均匀。
所有操作均需在无菌环境下进行，试剂瓶瓶盖可用70％乙醇擦拭，并自然干燥。
应使用预冷的移液器以保证Matrigel呈匀浆状。
细胞可在0.5mm厚度的Matrigel基质层表面生长，也可在1mm厚度的Matrigel三维基质内生长。
过度稀释的Matrigel会形成非胶质的蛋白层，可以用于细胞贴壁，但不能用于细胞的分化研究。
注意：可将冻融后的Matrigel分装在多个小管，所有分装均需用预冷的冻存管，迅速冷冻并保存，避免多次冻融。
Matrigel在22-35℃温度环境下快速成胶，因此溶解时在4℃冰上过夜冻融（4度时会随着温度的上升部分成胶）。
所有用品在使用前需置于冰浴，必须使用预冷的移液管、吸头及小管操作Matrigel。
成胶后的Matrigel可以在4℃24－48小时后重新呈液态。
注射步骤：1、注射前，必须保证Matrigel和细胞悬浮液在不冷冻的情况下，尽可能地保持低温，同时保证每一步都无菌操作。
2、对于每只注射小鼠，冰上混合0.5 mL细胞（2×105或更多细胞）和Matrigel混合液。
3、细胞注射体积需尽可能小。
通常，用含有2×106/mL细胞的250μL预冷培养基，混合250μL冰浴的Matrigel。
4、小鼠皮下注射，19G针用于组织样品，23G针用于培养的细胞。
注射动作必须快速，以避免Matrigel凝固。
5、抽出注射器时旋转以防止泄漏。
针头需要经常更换以避免堵塞。
用BD细胞回收剂（354253）或离散酶（354235）可降解Matrigel基质，冰浴7小时后回收得到细胞。
1.3人来源细胞外基质BD产品货号：354237储存和运输：-20度储存，干冰运输。
产品特性人源细胞外基质是来源于人体胚胎、经过层析纯化而得的细胞外基质提取物，由层粘连蛋白，IV型胶原，硫酸乙酰肝素蛋白多糖（HSPG）等成分组成。
人细胞外基质对贴壁依赖性上皮细胞特别是人源性细胞，有促进其贴壁、延展、代谢和分化的作用。
配方：0.02M磷酸钠溶液，pH值7.4包被方法：1用无血清培养基吸收细胞外基质到一定的浓度。
最终浓度应足以满足覆盖培养器皿表面的量。
例如：最终包被浓度为5.0 μg/cm2，应将基质稀释到50 μg /mL，在35 mm的培养皿中加入1mL基质，在60mm培养皿中加入3 mL基质。
2根据培养皿面积加入合适的已稀释基质。
3室温孵育2小时。
4吸走多余的基质。
5仔细清洗培养器皿，避免刮擦底部表面。
6包被后的培养器皿，应储存于2-8℃的潮湿环境中，或在无菌环境中自然干燥。
1.4Ⅲ型人重组胶原蛋白BD产品货号：354255储存和运输：-20度储存，干冰运输。
操作指南Ⅲ型胶原在皮肤、心血管系统和成年人的正常生理功能中发挥重要作用。
Ⅲ型胶原蛋白在正常人心血管纤维形成中起重要作用，还可用作促进细胞贴壁和调节细胞行为。
使用注意事项：Ⅲ型人重组胶原蛋白，可用于体外实验的薄胶包被，但也能优化用作胶。
如果一次不能使用完，可分装后储存于2-8℃。
推荐包被与成胶方法薄胶包被：1、选择合适的包被浓度。
推荐包被浓度为：0.2-2 μg/cm2，视细胞种类决定。
加入的包被体积必须足以覆盖细胞生长表面。
如有必要，可将胶原蛋白用10 mM醋酸稀释。
2、当生长表面完全覆盖后，室温孵育2小时。
倾斜培养皿至45度，使多余的胶原蛋白都聚集在培养皿的最低点。
3、用移液器去除多余的胶原蛋白。
4、在层流超净台上空气自然干燥或用灭菌的柔和气流干燥。
5、器皿包被完成，可备用。
成胶方法：1、加入9份的人重组胶原蛋白和1份的 10倍中性缓冲液或无菌组织培养基。
2、将混合液加入需要的培养器皿中。
3、室温孵育60分钟。
4、细胞可接种于胶的表面。
如需将胶从培养皿中转移或用于更多操作，在成胶前混合细胞和胶原蛋白。
1.5高浓度层粘连蛋白/巢蛋白混合物BD产品货号：354259储存和运输：-20度储存，干冰运输。
操作指南层粘连蛋白/巢蛋白混合物是EHS小鼠肿瘤基质膜的主要成分。
纯化后，成等摩尔浓度分布。
二价阳离子有利于混合物的形成。
在2 mg/mL浓度时，层粘连蛋白/巢蛋白可以形成三维胶，可用于细胞生长和分化的研究。
三维胶更接近体内微环境。
细胞的分化受到与单一或混合细胞外基质蛋白之间的相互作用的影响，因此在层粘连蛋白/巢蛋白混合物上培养细胞，可以用以观察并揭示细胞分化和功能性的特性机理。
混合蛋白的成胶过程对温度敏感。
此外，高浓度的层粘连蛋白/巢蛋白（2-6 mg/mL）也用于人下颌腺泡细胞分化和内皮细胞的血管形成研究。
注意事项：1、所有操作的工作温度不能高于4℃，蛋白需冰上操作保存，用冰块冷却的溶液或细胞悬浮液稀释。
2、由于混合蛋白溶液粘性较高，必须使用预冷的注射器型移液管，如Gilson M系列；避免使用气压式移液管或吸头。
推荐包被方法铺胶步骤：细胞在胶表面培养1、冰上溶解混合蛋白，所有操作均在冰浴上进行。
2、稀释到需要的浓度：稳定成胶浓度至少为3.5mg/mL。
3、用预冷的等压盐溶液或0.1mM钙离子培养基稀释4、将稀释的蛋白溶液加入到培养器皿，1-2小时，37℃成胶。
铺胶步骤：细胞在胶内部培养1、冰上溶解混合蛋白，所有操作均在冰浴上进行。
2、根据加样体积计算并稀释混合蛋白，成胶浓度至少为3.5mg/mL，稀释过程需在预冷的培养器皿或小管中进行。
3、加入预冷的细胞悬液，用等压盐溶液或培养基调节溶液浓度到第二步所需的浓度。
4、用Gilson M 系列移液管混匀混合蛋白和细胞悬液。
5、将混合后的细胞蛋白悬液加入到预冷的培养板中，37℃孵育1-2小时成胶。