AAV-LC3操作手册
VAV-Compact模块式控制设备用户手册说明书
VAV-CompactVAV-CompactContentsModbus General Notes 2 Modbus Register Overview 3 Modbus Register Description 4Volumetric flow compact control deviceModbus General NotesGeneral information Date25.03.2019Product Name VAV-CompactActuator type..MV-D3-MOD, LHV24A-D3-MODProtocol Modbus RTU over RS-485Modbus RTU Transmission formats1-8-N-2, 1-8-N-1, 1-8-E-1, 1-8-O-1 (Default: 1-8-N-2)Baud rates9‘600, 19‘200, 38‘400, 76‘800, 115’200 Bd(Default: 38‘400 Bd)Address 1...247 (Default: 1)Number of nodes Max. 32 (without repeater)Terminating resistor120 �Parameterisation Tool ZTH EUQuick adressing Actuators support quick addressing via the …Address“ and …Adaption“ buttons.For detailed, information please see product datasheet (chapter Service).Register implementation All data is arranged in a table and addressed by 1..n (Register No.) or 0..n-1 (Address). No distinctionis made between data types (Discrete Inputs, Coils, Input Registers and Holding Registers). As aconsequence, all data can be accessed with the two commands for Holding Register. The commandsfor Discrete Inputs and Input Registers can be used as an alternative.Commands Standard commands:Read Holding Registers [3]Write Single Register [6]Optional commands:Read Discrete Inputs [2]Read Input Registers [4]Write Multiple Registers [16]Command …Read Discrete Inputs“The command reads one or more bits and can alternatively be used for Register No. 105(Malfunction and Service information).Example The start address to be used is 1664 → 104 (Register Address) * 16 (Bit) = 1664 Interpret values in the registers All values in the register are unsigned integer datatypes.Example Read (Function 03, 1 Register) Value Register No. 12 = 0001‘1010‘1100‘10002 = 6‘85610Actual Value = Value * Scaling factor * Unit = 6‘856 * 0.01 * m3/h = 68.56 m3/h 32-Bit values in two registers Values that exceed 65,535 are stored in two consecutive Registers and have to be interpreted as…little endian“ / LSW (Least Significant Word) firstExample Register No. 10 (AbsFlow LowWord) = 14,55110 = 0011‘1000‘1101‘01112Register No. 11 (AbsFlow HighWord) = 1910 = 0000‘0000‘0001‘00112AbsFlow HighWord AbsFlow LowWord1914,5510000‘0000‘0001‘001120011‘1000‘1101‘01112AbsFlow = 0000‘0000‘0001‘0011‘0011‘1000‘1101‘01112 = 1,259,73510 = 1259.735 l/hMath formula:AbsFlow = ( AbsFlow HighWord * 65,536 ) + AbsFlow LowWordAbsFlow = ( 19 * 65,536 ) + 14,551 = 1,259,735 = 1259.735 l/hDeactivated registers If a register is not supported by a device or by a device setting it is indicatedwith 65‘535 (1111‘1111‘1111‘11112).!All writeable registers on registers >100 are persistent and are not supposed to be written on aregular base.Operation No.Address Register Access 10Setpoint [%]R / W21Override control R / W32Command R / W43Actuator type R54Relative position [%]R65Absolute position [°] [mm]R76Relative volumetric flow [%]R87Absolute volumetric flow [m3/h]R98Sensor value 1 [mV] [–]R109––1110Absolute volumetric flow in unit selected LowWordR1211HighWord1312Setpoint analog [%]RService No.Address Register Access 10099Bus termination R101100Series number 1st partR 102101Series number 2nd part103102Series number 4th part104103Firmware version R105104Malfunction and service information R106105Min [%]R / W107106Max [%]R / W108107Sensor type 1R / W109108Bus fail position R / W110109Communication Watchdog R / W111110Nominal volumetric flow [m3/h]R112111––113112Nominal volumetric flow in unit selected LowWordR114113HighWord115114––116115––117116Control Mode R / W 118117Unit Selection Flow R / W 119118Setpoint source R / WNo.Address DescriptionComment RangeEnumerationUnit Scaling Access10SetpointSetpoint for actuator between Min (Register No. 106) and Max (No. 107)0…10‘000Default: 0%0.01R / W21Override ControlOverride setpoint with defined values 0: None1: Open2: Close3: Min4: Mid5: MaxDefault: None(0)––R / W32CommandInitiation of actuator functions for service and testAfter command is sent, register returns to None(0)With Reset(4) all Malfunction and Service Information (Register No. 105)Information can be reset.0: None1: Adaption2: Test3: Sync4: ResetDefault: None(0)––R / W43Actuator Type0: Actuator not connected1: Air / Water2:VAV / EPIV3: Fire4: Energy Valve5: 6way EPIV––R 54Relative Position0...10‘000%0.01R65Absolute PositionThe unit depends on the device:[°] for actuators with rotary movement[mm] for actuators with linear movement 0...max angle / stroke°mm11R76Relative volumetric flowRelative volumetric flow of Vnom0...10‘000%0.01R87Absolut volumetric flow0...Vnom m3/hPa1R98Sensor 1 ValueCurrent value of sensor 1, depending on the setting of the Sensor 1 Type(Register No. 108)[mV] if Sensor 1 Type (Register No. 108) is Active(1)[0 / 1] if Sensor 1 Type (Register No. 108) is Switch(4)0...65‘535mV0 / 11R109–––––1110Absolute volumetric flowAbsolute flow in unit selected (Register No. 118) LowWordLower 16 bit of 32 bit value–UnitSel0.001R1211Absolute volumetric flowAbsolute flow in unit selected (Register No. 118) HighWordUpper 16 bit of 32 bit value1312Setpoint AnalogShows the setoint in % if actuator is control by analog signal0...10‘000%0.01RNo.Address DescriptionComment RangeEnumerationUnit Scaling Access10099Bus TerminationIndicates if bus termination (120 Ω) is enabledBus termination can be set with the configuration tools 0: inactive1: activeDefault: inactive(0)––R / W101100Series Number 1st partEach device has an unambiguous series number, which is either impressedon or glued to the housingThe series number consists of 4 segments, although only parts 1, 2 and 4are displayed on ModusExample: 00839-31324-064-0081st part: 008392nd part: 313244th part: 008–––R102101Series Number 2nd part–––R 103102Series Number 4th part–––R 104103Firmware VersionFirmware version of communication moduleExample: 302, Version 3.02–––R105104Malfunction and Service InformationValue is bit-coded. More than one bit can be set to 1All bits not montioned in the enumeration are not used for this actuator rangeMechanical travel increased: The actuator has been moved outside theadapted working rangeActuator cannot move: Mechanical overload e.g. blocked actuator, etc.Internal activity: Actuator performs a test run, adaption, etc.Gear disengaged: The gear disengaged button is pressedBus Watchdog triggered: Timeout for the Bus Watchdog expired Bit1: Mech travel increasedBit2: Actuator cannot moveBit8: Internal activityBit9: Gear disengagedBit10: Bus Watchdog triggered––R106105VminMin has to be ≤ Max 0...VmaxDefault: 0%0.01R / W107106VmaxMax has to be ≥ Min and > 20%Vmin...10‘000Default: 10‘000%0.01R / W108107Sensor 1 TypeIf Setpoint Source (Register 119) is analog (Hybrid mode) the sensor type 1can be set to Active(1) in order to see the Setpoint Analog in mV 0: None1: Active / Hybrid2: –3: –4: SwitchDefault: None(0)––R / W109108Bus Fail PositionModbus communication is not monitored as standard. In the event of abreakdown in communication, the actuator retains the current setpointThe bus implementation tracks the Modbus communication. If neitherthe Setpoint (Register No. 1) nor the Override Control (Register No. 2) isrenewed before the Timeout for Bus Watchdog (Register No. 110) expires,the actuator controls to the Bus Fail PositionTriggered bus watchdog is indicated in the Malfunction and ServiceInformation (Register No. 105)0: None / Last setpoint1: Fast close2: Fast open3: Mid position(parameterized)Default: None(0)––R / W110109Timeout for Bus Watchdog in sTime until Bus Fail will be detected. If Bus Watchdog = 0 then deactivatedIf Bus Fail Position (Register No. 110) different from 0, the Bus Fail Positionbecomes active after the Timeout for Bus Watchdog is expired 0...3‘600Default: 0If Bus Fail Position (RegisterNo. 110) not None(0), thenDefault: 120s1R / WNo.Address DescriptionComment RangeEnumerationUnit Scaling Access111110Nominal volumetric flow–m3/h1R 112111–––––113112Nominal volumetric flowVnom in unit selected (Register 118) LowWordLower 16 bit of 32 bit value–UnitSel0.001R114113Nominal volumetric flowVnom in unit selected (Register 118) HighWordUpper 16 bit of 32 bit value115114–––––116115–––––117116Control Mode0: Position control1: Flow control––R / W118117Unit Selection Flow0: m3/s1: m3/h2: l/s3: l/min4: l/h5: gpm6: cfmDefault: m3/h(1)––R / W119118Setpoint SourceAnalog: Setpoint from analog signal 0 ... 10 V on wire 3Bus: Setpoint from Modbus (Register 1)0: Analog1: BusDefault: Bus(1)––R / W。
SYW95A-V3门禁管理系统进阶操作手册
21 控制群组为 On
22 控制群组为 Off
23 开门逾时
24 门区强行进入
25 反胁迫
侦测功能说明 读卡辨识 读卡辨识 读卡辨识 读卡辨识 读卡辨识 读卡辨识 读卡辨识 读卡辨识及按键输入侦测 读卡辨识 读卡辨识 按键输入侦测 数字输入点侦测 数字输入点侦测 数字输入点侦测 数字输入点侦测 数字输入点侦测 数字输入点侦测 控制群组之动作时区侦测 控制群组之动作时区侦测 数字输入点侦测 数字输入点侦测 按键输入侦测
SYW95A-V3 门禁管理系统 进阶操作手册
Version 2.0
© 2005 by SYRIS Technology Corp.
1
1.关于流程控制 (Flow Control).................................................................................................................3 1.1 控制器需侦测之事件对照表(Event) ............................................................................................4 1.2 控制器可响应之动作对照表(Action) ...........................................................................................5 1.3 设定流程控制 ................................................................................................................................6 1.4 系统预设流程控制对照表 (Default Flow Control) .....................................................................8
MSI-SR-LC31安全门控制器说明书
Technical dataBasic dataSeries MSI-SR-LC31Application Base device for E-Stop and safety door applicationsFunctionsFunctions Monitoring of position switchesMonitoring of solenoid switchesMonitoring of optoelectronic protective devicesMonitoring of proximity switchesMonitoring of E-Stop circuitsRestart AutomaticCharacteristic parametersSIL3,IEC 61508SILCL3,IEC/EN 62061Performance Level (PL)e,EN ISO 13849-1PFH D0.00000003 per hourPFH D3E-08 per hourMission time T M20 years,EN ISO 13849-1Category4,EN ISO 13849Electrical dataSupply circuitNominal voltage U N24 V AC/DCNominal frequency50 ... 60 HzRated control supply voltage U S at AC 60 Hz20.4 VRated control supply voltage U S at AC 50 Hz26.4 VMax. rated control supply voltage at AC 50 Hz26.4 VMin. rated control supply voltage U S at AC 50 Hz20.4 VMin. rated control supply voltage U S at DC20.4 VMax. rated control supply voltage at DC26.4 VMin. rated control supply voltage at DC20.4 VRated power AC 2.9 V·ARated power DC 1.6 WGalvanic isolation between supply and control circuit Yes (when U N≥ AC 42-48 V, AC 115-230 V, AC 230 V)Number of outputs, safety-oriented, undelayed, contact-based 3 Piece(s)Number of outputs, signaling function, undelayed, contact-based 1 Piece(s)Release current paths NOSignaling current paths NCContact material Ag alloy, gold-platedUsage category AC-15 (NO contact)Ue 230V, Ie 3AUsage category DC-13 (NO contact)Ue 24V, Ie 3AShort circuit protection (NO contact)gG class safety fuse 6A, melting integral Nominal switching voltage, release current paths AC230 VNominal switching voltage, signaling current paths AC230 VMax. thermal continuous current I th, release current paths8 AMax. thermal continuous current I th, signaling current paths 5 AMax. total current I² of all current paths25 A²Mechanical life time100,000,000 switching cyclesControl circuitEvaluation of the inputs Two-channelNominal output voltage DC24 VInput current at the control inputs (safety circuit/reset circuit)25 mAMax. peak current at the control inputs (safety circuit/reset circuit)100 mAMax. cable resistance, per channel≤ (5 + (1.176 x U B/ U N- 1) x 100) ΩMinimum switch-on time100 msResponse time (automatic start t A2)350 msResponse time (manual start t A1)350 msTest pulse time permitted t TP 1 msRelease time t R10 msRecovery time t W750 msConnectionNumber of connections 1 Piece(s)Connection 1Type of connection TerminalFunction Voltage supplySignal INSignal OUTType of terminal Screw terminalNo. of pins16 -pinCable propertiesConnection cross sections 2 x 0.2 to 1.0 mm², wire1 x 0.25 to 2.5 mm², wire with wire-end sleeve1 x 0.2 to 2.5 mm², wire1 x 0.2 to 2.5 mm², wire2 x 0.25 to 1.0 mm², wire with wire-end sleeve2 x 0.2 to 1.0 mm², wireMechanical dataDimension (W x H x L)22.5 mm x96.5 mm x114 mmNet weight210 gHousing color GrayType of fastening Snap-on mountingCertifications c UL USTÜV RheinlandClassification*********27371819*********27371819 ETIM 5.0EC001449Dimensioned drawingsAll dimensions in millimetersElectrical connectionConnection 1Type of connection TerminalConnection 1Function Voltage supplySignal INSignal OUT Type of terminal Screw terminal No. of pins16 -pinTerminal Assignment13Release current path 1 (NO contact)14Release current path 1 (NO contact)23Release current path 2 (NO contact)24Release current path 2 (NO contact)33Release current path 3 (NO contact)34Release current path 3 (NO contact)41Signaling current path (NC contact)42Signaling current path (NC contact)A1+24VA2GNDS11Control circuit 1S12Control circuit 1S21Control circuit 2S22Control circuit 2S34Control circuit of reset buttonS52Control circuit 2Circuit diagramsNotesObserve intended use!•The product may only be put into operation by competent persons.•Only use the product in accordance with its intended use.。
艾克莱·高压探头和测量仪器说明书
Item shipped may vary from model p ictured here.Model 1600A High Voltage ProbeMAXIMUM INPUT: 40kV DC or 28kV AC, 50/60Hz.INPUT RESISTANCE: 1000M W .DIVISION RATIO: 1000:1 (into 10M W ).For use with: DMMsModel 1651 50 Ampere Shunt:E xternal 0.001W ± 1%, 4-terminal shunt; extends current mea s ur i ng capability of Keithley DMMs to 50A.CABLE LENGTH: 1.4m (56 in).For use with: DMMsItem shipped may vary from model p ictured here.Model 1681 Clip-On Test Lead Set: Two 1.2m (48 in) leads terminated with banana plugs and spring action clip-on probes.For use with: DMMsItem shipped may vary from model p ictured here.Model 1751 Safety Test Leads: 91.4cm (36 in) test lead set supplied with each Model 175A and 197A. Finger guards and shroud e d banana plugs help minimize the chance of making contact with live circuitry.For use with: All DMMs, Series 2400 SourceMeter ®SMU InstrumentsModel 1752 Premium Safety Test Lead Kit:Includes two banana leads with safety shrouds on both ends, two fully insulated alligator clips, two fully insulated needle probes, and two spring-action clip-on probes with safety blocks, in a carrying case.For use with: All DMMs, Series 2400SourceMeter SMU InstrumentsA C C E S S O R I E SItem shipped may vary from model p ictured here.Model 1754 Universal Test Lead Kit: 10-piece test lead kit with interchangeable plug-in acces-sories. Kit includes: 1 set of test leads, 91.4cm (36 in), 1 red and black; 2 spade lugs, 2 standard banana plugs, 2 hooks, and 2 min i a t ure alligator clips with boots.For use with: All DMMs, Series 2400SourceMeter SMU InstrumentsModel 2187-4 Low Thermal Test Lead Kit: Includes an input cable with banana termina-tions, banana extensions, sprung-hook clips, alligator clips, needle probes, and spade lugs to connect the Model 2182A to virtually any DUT. The kit is also used to connect the Model 2182A directly to the Model 622X guard to improve response time for pulsed measure-ments or to reduce errors when measuring high impedance devices.For use with: 2182A, 6220, 6221, 6220/2182A,6221/2182AModel 2600-BAN: 1m (3.3 ft) banana test leads/adapter cable for a single Series 2600B SourceMeter channel (two needed for use with Model 2602B and 2612B). Provides safety banana connections to Hi, Sense Hi, Lo, Sense Lo, and guard.For use with: 2601B, 2602B, 2611B, 2612BModel 2260-008: Test lead set with lugs, 10AWG, 122m (48 in), red/blue wire pair with #10 terminal lugs on each end.For use with: Series 2260BModel 2260B-EXTERM: Extended Terminal Test Leads, 0.7m (28 in.), and terminal box to bring outputs to the front of the instrument or another location. Magnetic base attaches to side of the instrument.For use with: Series 2260BModel 3706-BAN: 1.4m (4.6 ft) banana test leads/adapter cable for Model 3706A mainframe with high performance multimeter option. Provides direct connection to high performance multimeter utilizing rear panel backplane con-nector. Safety banana connections to HI, LO, Sense HI, Sense LO, and AMP.For use with: Series 3700A mainframesModel 3706-TLK Test Lead Kit: Includes 3706-BAN banana test leads/adapter cable and plug-in test lead accessories. Test lead accessories include: 4 spring hook probes, 2 needle test probes, 4 safety plug adapters, and carrying case.For use with: Series 3700A mainframesModel 5804 Test Lead Set: Designed to be used with Keithley instruments that measure 4-terminal resistance. Contains: 2 test leads, 0.9m (36 in), red and black; 2 test clips, red and black; 2 plunger clip adapters, red and black; 2 alligator clips with boots (accept standard test probe tip); 2 alligator clips with boots (barrels accept standard banana plugs).For use with: Series 2400 SourceMeter ® SMUInstruments, 2750, DMMsItem shipped may vary from model p ictured here.T e s t l e a d s a n d p r o b e sModel 5805 Kelvin Probes: 2 spring-loaded test probes, red and black, 0.9m (36 in) with banana plug termination. De s igned to be used with Keithley in s tru m ents that measure 4-terminal re s is t ance and in-circuit current. 1⁄8-in spacing between pin tips. A package of 8 re p lace m ent contacts (P/N CS-551) is available.Model 5805-12: Similar to 5805 but 3.6m (12 ft) in length.For use with: Series 2400 SourceMeter SMUInstruments, 2750, DMMsModel 5806 Kelvin Clip Lead Set: 2 clip test leads, red and black, 0.9m (36 in) with banana plug ter m i n a t ion. Designed to be used with Keithley instruments that meas u re 4-terminal re s is t ance. Maximum jaw opening ½ in. A package of 8 r eplace m ent elastic bands (P/N 5806-306B) is a vailable.For use with: Series 2400 SourceMeter SMUInstruments, 2750, DMMsModel 5807-7 Helical Spring Point Test Leads: Two excellent Kelvin test probes for Keithley in s tru m ents that measure 4-terminal re s is t ance. The probes have point e d tips that rotate when pressure is exerted on the handle. Good contact is assured, and the handle can be held at any angle to the mea s ure m ent surface. The lead wires ter m i n ate in banana plugs. Lead wire length: 2m (7 ft).For use with: Series 2400 SourceMeter SMUInstruments, 2750, DMMsModel 5808 Low Cost, Single Pin, Kelvin Probes: Two 1.17m (46 in) long, single fixed pin probes with source and sense leads per probe and color coded standard safety banana plugs. Designed for applications requiring Kelvin resistance measurements with the convenience of a single point of circuit contact. The Kelvin connection at the base of each probe pin results in very low resistance errors, eliminating all wire and probe body resistance. For the highest pre-cision sensing applications, use the Model 5805, 5806, or 5809 for complete control over sensing point placement.For use with: 2700, 2701, 2750, Series 2400,DMMsModel 5809 Low Cost, Kelvin Clip Lead Set: Two 0.94m (37 in) long probes with alligator style clips, each with opposing blade source and sense connections to color coded standard safety banana plugs. Designed for applications requiring Kelvin connection to wires, axial lead-ed components, and other terminals able to be clipped onto. The miniature blade design accom-modates narrow available width applications as well as general lighter duty applications. The maximum jaw opening is 9.14mm (0.36 in) and the blade width is 2.92mmm (0.115 in).For use with: 2700, 2701, 2750, Series 2400,DMMsModel 6517-RH Humidity Probe: The 6517-RH comes with a 3m (10 ft) extension cable and is designed to be used with the 6517B Electrom-eter. The 6517-RH measures the relative humidity of the testing environment.OPERATING TEMPERATURE RANGE: –10°C to +60°C OPERATING HUMIDITY RANGE: 0 to 100% RH MEASURING RANGE: 10 to 90% RHTEMPERATURE DEPENDENCE: <±2% from –10°C to +60°CFor use with: 6517A and 6517B ElectrometersA C C E S S O R I E SModel 6517-TP Thermocouple Bead Probe: Designed to be used with the 6517B Electro m -eter, the 6517-TP measures the temperature of the testing en v i r on m ent. 6517-TP is 36 in. long and has a temperature calibration (range) of 0°C to 1250°C. Type K thermocouple.For use with: 6517A and 6517B Electrometersand 2110 DMMModel 7401: Type K ther m o c ou p le wire kit includes 30.5m (100 ft) of type K (chromel-alu-mel) ther m o c ou p le wire.For use with: 2001-TCSCAN, 2110, 3720, 3721,3724, 7057A, 7700, 7706, 7708Model 8605 High Per f or m ance Modular Test Leads: Highest quality lead with sili c one insula-tion, and removable test probes. Rated to 1500V. UL listed.LENGTH: 1m (3.3 ft).For use with: All DMMs, Series 2400 SourceMeter ® SMU InstrumentsItem shipped may vary from model p ictured here.Model 8606 High Per f or m ance Modular Probe Kit: For use with Model 8605 High Per-for m ance Modular Test Leads. Contains 2 each spade lugs, alligator clips, and re t ract a ble hooks.For use with: All DMMs; Series 2400 and Series 2600B SourceMeter SMU Instruments;Series 3700AModel 8608 High Per f or m ance Clip Lead Set: High quality clip lead set with removable test clips. NRTL listed. Rated to 1000V CAT 3 and 20 amps.LENGTH: 1.2m (3.9 ft).For use with: 2450, Series 2400 SourceMeterSMU Instruments, Series 3700A DMMsModel 8681 Low Cost RTD: This inexpensive RTD has small size (0.09 in × 0.09 in) and fast thermal response time. 100W , 0.385%/°C ele-ment. Meets or exceeds DIN 43760-1980-A, IEC 751:1983-A, BS1904: 1984-A, and JIS C1604-1981-0.2. Can be used with Model 8680 RTD Input Probe Adapter. Measures from –200° to +600°C.LENGTH: 0.9m (3 ft).ACCURACY: ±(0.15°C + 0.002 × |t|)/°C, where |t| is absolute value of temperature in °C.For use with: 2001, 2002, 2010 DMMs (with 8680), 7700, 7702, 7706, 7708Item shipped may vary from model p ictured here.Model CA-109A Test Lead Set: For output con-nec t ions (two red, two black).For use with: 2000-SCAN, 2001-SCAN, 2001-TCSCANT e s t l e a d s a n d p r o b e s。
艾礼安ABIV3.0红外栅栏说明书
ABI AIR系列主动红外电子光栅产品说明书V2020.10.16一、概述红外光栅，又叫电子光栅或红外栅栏，是主动红外探测器的一种。
采用变频互射技术，一旦有人员或物体挡住了光栅主机和光栅从机之间的红外光束时（出厂默认为遮挡相邻两束及以上光束），立即输出报警信号。
二、产品特性封闭式外接线结构设计（已申请专利），创新性地吸取“瓶盖”倒置设计灵感，在保留外接线的同时，将光栅的防水性能提升到了新的高度；采用数字变频技术与32位高速MCU控制，整体稳定性更好，可靠性更高；采用高档铝合金外壳，防拆、防剪、防移动功能设计；支持异步、同步两种工作模式(出厂默认为异步工作模式)；采用变频互射技术，两段频率可选(出厂默认为A频工作模式)，彻底解决相邻红外光栅间相互干扰，避免了太阳光、手电筒、车灯的的直射问题。
支持双束识别功能，能有效地防止小动物、飞鸟等引起误报；灵敏度高、防范警戒距离三档可调；抗干扰能力强：可在风、霜、雨、雪、雾、潮、太阳光斜照的室外恶劣环境下使用；独立防拆开关，设计更合理可靠，确保光栅不受拆卸破坏；支持蜂鸣器校准提示；已通过了3C、CE、RoHS认证；三、技术参数警戒距离：同步工作0-100米，异步工作10-80米光束：2/4/6/8/10/12光束电源电压：DC 9~30V消耗电流：50-120mA环境温度：-30~70℃报警输出：常开、常闭可选(出厂默认常闭)，接点容量30V 2A 探测方式：遮挡相邻两光束（可设置单光束报警）报警持续时间：≥1S（可设置即时报警）独立防拆输出：当外壳移开时，常闭输出反应时间：≤40ms光轴调整角度：水平180°、垂直不可调其它附加功能：LED指示、蜂鸣提示材质：铝材、PC工程塑料四、产品结构五、安装及使用说明1、安装前，如需对光栅的报警输出状态或报警模式等进行设置，请先确保光栅断电，打开主机管塞（左右轻轻摇动管塞后更易打开），拉出主机主板，进行设置（请勿过份用力，以防电路板折断)。
LC-B3标准说明书
目录一、概述 (2)二、主要特点及技术参数 (2)三、接线端子说明 (4)四、面板操作说明 (5)1、操作面板说明 (6)2、控制面板功能 (7)3、规范参数的设定及变更 (7)4、多规范调用的设定 (7)5、内存参数清除 (7)五、控制箱的内部设定 (8)1、电源设定 (8)2、内部拔位开关的设定及说明 (8)六、互锁装置 (9)1、内部电路图 (9)2、两台控制器互锁装置接线图 (9)3、两台以上控制器互锁的示意图 (10)七、安装尺寸 (10)八、电气原理图 (11)九、控制器接线图 (12)一、概述LC-B3是在SKM1基础上研发的新型电阻焊控制器,具有直观易操作等特点。
主要用于单相交流，次级整流点（凸）焊机及对焊机的控制，每个循环可以完成从加压，缓升，焊接1，冷却1，焊接2，维持，休止程序。
二、主要特点及技术参数1、具有电网电压波动补偿，减少电网不稳定对焊接质量影响。
2、具有次级恒流控制功能，避免工件表在面状态对焊接质量的影响。
3、具有电流缓升功能，以达到各种的焊接工艺的要求。
4、具有锻压功能。
5、可存贮7套焊接规范。
6、可以利用控制面板直接对各焊接规范参数设置，操作方便，调整简单。
7、两台机可直接进行通电互锁，或与本公司生产分时控制器相连，可与多台设备进行互锁。
8、具有故障自诊断功能，并能显示，方便检修。
10、外形尺寸：（W）345×（D）110×（H）32211、重量：6公斤12、设备环境：a、电压和频率1）、焊接电源供给：单相AC380V /220V±10%，50/60H Z2)、电磁阀电压：AC220V（可选24V，由供方设定）3）、电源消耗：少于15W（待机状态）b、外环境1）、环境温度：0-400C2）、相对湿度：小于或等于90%没有露水c、冷却条件1）、控制器：自然冷却2）、可控硅：水冷入水口温度：≤300C入水量：≥ 3L/m电阻率：≤5000Ω/cm三、接线端子说明四、面板操作说明1、操作面板说明2、控制面板功能●工作开关：该开关为本控制器的电源开关，使用时置于“ON”，系统上电。
岛津LCAVP液相色谱仪键盘操作PPT教案学习
光谱扫描操作
9、注入样品 10、当被扫描的色谱峰开始出峰时，关闭泵 11、按SHIFT，3(SCAN)键，开始扫描至结束
输出光谱图 12、确认数据处理机信号线连接于检测器的RECORDER
端口 13、设定数据处理机衰减（ATTEN）为3或4 14、设定绘图速度，[PLOT SPEED] 15、输出扫描光谱图，[SPC PLOT] , ENTER
第54页/共34页
输液泵时间程序设定
1、在初始屏幕下，按 EDIT 键，显示
USED LEFT 0 320
2、按 Enter 键，显示
TIME FUNC VALUE 0.00 ～ 999.99 min
3、输入时间，如2（分钟）。按 2 ，Enter 4、重复按 func 键至显示所要设定的命令 5、输入命令参数值 6、按 CE 键，退出时间程序编辑状态
设定双波长检测时的 的输出模式 开/关氘灯 显示氘灯累积使用时间
设定系统控制器地址
第2109页/共34页
检测器参数设定
LOCAL 0: Remote 1: Local
WAVE CHECK Enter to Start
RT RNG 10 Input 2 ~ 10000
RT THR 0.0001 Input 0.0001 ~ 1
SPD－10AVP
S.V. zero
Z.Shift
shift
prog.run remote pol power
run
mark VP
del
dual
CE
7
8
edit
back
4
5
func
1
2
0
.
第165页/共34页
迪亚尔brand电子厨Sink 50N 32使用指南与维护指南说明书
Instructions for use and maintenance of gas, gas-electric and electric cookers. Series:32-Thank you for choosing one of our quality products, capable of giving you the very best service. To make full use of its perfor-mance features, read the parts of this manual which refer to your appliance carefully. The Manufacturer declines all responsibility for injury or damage caused by poor installation or improper use of the appliance.-To ensure its appliances are always at the state of the art, and/or to allow constant improvement in quality, the manufacturer reserves the right to make modifications without notice, although without creating difficulties for users.-When ordering spare parts, inform your dealer of the model number and serial number punched on your appliance’s nameplate, visible inside the warming compartment or on the back of the cooker.-APPLIANCE COMPLYING WITH THE FOLLOWING DIRECTIVES:- EEC 90/396- EEC 73/23 and 93/88-EEC 89/336 (radio-frequency inter-ference)- EEC 89/109 (contact with foods)FOREWORD-Refer only to the headings and sections covering accessories actually installed on your cooker.Technical data andspecifications.............................33Installation..........................34 - 36Ventilation......................................34Positioning.....................................34Gas connection .............................34Adapting to different typesof gas ............................................35Replacing the injectors ..................35Regulating the air ..........................35Minimum setting ............................35Electrical connection .....................36Safety device (36)For the user....................... 37 - 40Ventilation......................................37Igniting the burners....................... 37Igniting the gas oven .................... 37Igniting the gas grill .......................37Safety device.................................37Using the gas hob .........................37Using the electric hot-plates ..........37Using the gas oven........................38Using the gas or electric grill ........38Using the conventionalelectric oven ..................................38Using the conventionalmultifunction electric oven .............39Oven with thermostat ....................39Warming compartment ..................39Advice and precautions.. (39)Figures .............................. 41 - 43ELECTRIC HOTPLATESø 1451,0 kW - Normal hotplate1,5 kW - Rapid hotplateø 1801,5 kW - Normal hotplate2,0 kW - Rapid hotplate ELECTRIC OVEN POWEROven: (1,1kW bottom0,7kW top element)2,2 kW Grill:2,0 kW Cat.: see nameplate on cover; Class 1 or 2.1 Type “X” cookersEQUIPMENTAll models are equipped with safety device for oven and grill burners.Depending on the models, cooker may also have:-Oven thermostat (or tap)-Electric oven lighting-Grill burner or element-One or more electric hotplatesFor the LAYOUT OF HOB BURNERS see the models illustrated in figure 1 at the back of this manual.For the ELECTRIC WIRING DIAGRAM see figure 2 at the back of this manual.The electrical power is stated on the name-plate visible inside the warming compartment (if present) or on the back of the cooker.A copy of the nameplate is glued to the coverof this manual (for gas or gas-electric prod-ucts only).33INSTALLATIONThe appliance must be installed by qualified staff working in accordance with the regula-tions in force.Before installing, ensure that the appliance is correctly preset for the local distribution conditions (gas type and pressure).The presettings of this appliance are indi-cated on the nameplate shown on the cover. This appliance is not connected to a flue gas extractor device. It must be installed and connected in accordance with the regulations in force.This appliance may only be installed and may only operate in rooms permanently ventilated in accordance with national regulations in force.VENTILATIONThe rooms in which gas appliances are in-stalled must be well ventilated in order to al-low correct gas combustion and ventilation. The air flow necessary for combustion is at least 2 m3/h for each kW of rated power. POSITIONINGRemove the packaging accessories, includ-ing the films covering the chrome-plated and stainless steel parts, from the cooker. Position the cooker in a dry, convenient and draft-free place. Keep at an appropriate dis-tance from walls which may be damaged by heat (wood, linoleum, paper, etc.).The cooker may be free-standing (class 1) or between two units (in class 2 st 2-1) the sides of which must withstand a tempera-ture of 100°C and which must not be higher than the working table.CONNECTING TO THE GAS SUPPLY Before connecting the cooker, check that it is preset for the gas to be used. Otherwise, make the conversion as described in the section headed “Adapting to different gas types”. The connection is on the right; if the pipe has to pass behind the cooker, it must be kept low down where the temperature isabout 50°C.-Rigid connection (see fig. 3 A + B)The connection to the mains gas supplymay be made using a rigid metal pipe orwith a metal hose. Remove the hose con-nector (if already fitted) and screw the rigidunion onto the threaded connection of thegas train (see fig. 3A). The union for rigidconnection may already be fitted on the gastrain, or may be amongst the cooker ac-cessories. Otherwise, it can be obtainedfrom your dealer.If national regulations permit, a metal hosecomplying with the national standards canbe screwed directly onto the threaded con-nection of the gas train, fitting a seal (seefig. 3B). However, users are strongly rec-ommended always to fit the rigid union.-Connection using a rubber hose (see fig.3C). (For butane/propane gas only).Connect a rubber hose carrying the con-formity mark currently in force to the hoseconnector. The hose must be replaced atthe date indicated at the latest, and mustbe secured at both ends using stand-ard hose clamps. It must be absolutelyaccessible to allow its condition to bechecked along its entire length.CAUTION:-Use of the hose connector is only per-mitted for free-standing installation. Ifthe appliance is installed between twoclass 2 st. 2-1 unions, the rigid union isthe only form of connection permitted.IMPORTANT:-After installation, check that the connec-tions are airtight.-For operation with butane/propane, checkthat the gas pressure is as indicated on thenameplate.-Use only standard rubber hoses. For LPG,use a hose which complies with the nationalregulations in force.-Avoid sharp bends in the pipe and keep itwell way from hot surfaces.References to the regulations covering the gas connection to the appliance: ISO 7-1.34ADAPTING TO DIFFERENT TYPES OF GASIf the cooker is not already preset to operate with the type of gas available, it must be con-verted. Proceed as follows:-Replace the injectors (see table on page 33);-regulate the primary air flow;-regulate the minimum settings.N.B.: every time you change the type of gas, indicate the new type of gas on the serial number label.REPLACING THE HOB BURNER INJEC-TORS (fig. 4)-Remove the lid of the cooker by lifting it off its supports;-remove the grids, burner caps and burn-ers, lifting them off;-unscrew the 2 screws (above) or nuts (be-low) at the back which secure the work top, and pull it out forward.-remove the mixer pipes and replace the injectors using a 7 mm socket wrench. REPLACING THE OVEN BURNER INJEC-TOR (Fig. 5)-Loosen the screw which secures the bot-tom of the oven;-remove the oven bottom (pulling it forward); -remove the oven burner, after taking out the screw which secures it;-replace the injector using a 7 mm socket wrench.REPLACING THE GRILL BURNER INJEC-TOR (Fig. 6)-Remove the burner after taking out the two screws which secure it;-replace the injector using a 7 mm socket wrench.IMPORTANT:-Never over-tighten the injectors;-after replacing, check that all the injectors are airtight.Burner G20 20mbar G30 28-30mbarG31 37mbar Auxiliary3 4Semi-rapid3 3Rapid4 6Oven- -Grill3 8REGULATING THE BURNER AIRRefer to the table below (indicative values) for regulation of the gap H in mm (fig. 4 for the hob, fig. 6 for the grill).Check operation of the burner:-Ignite the burner at maximum flame;-the tongue of the flame must be clear andwith no yellow tip, and must adhere closelyto the burner. If too much air is supplied,the flame detaches from the burner andmay be dangerous. If the air supply is in-sufficient, the flame has a yellow tip andsoot may form.SETTING HOB BURNER MINIMUM LEV-ELSIf the cooker is to work on bottled gas (bu-tane/propane), the tap by-pass must be screwed right down.The cooker may be equipped with type A taps, with by-pass inside (accessed by in-serting a small screwdriver into the rod) or type B taps, with by-pass on the outside on the right (accessed directly). See figure 7.If the cooker is to work on natural gas, pro-ceed as follows for both types of tap:-Ignite the burner at maximum flame;-pull off the knob, without using a leveragainst the control panel, which might bedamaged;-access the by-pass with a small screwdriverand back off by about 3 turns (turning thescrewdriver anti-clockwise);-turn the tap rod anti-clockwise again until itstops: the burner will be at maximum flame;-screw the by-pass slowly back in, withoutpushing the screw-driver, until the flame hasapparently shrunk to 1/4 of the maximumsize, checking that it is sufficiently stable35even in quite strong draughts.SETTING OVEN BURNER MINIMUM LEV-ELSIf the cooker is to work on bottled gas (bu-tane/propane), the thermostat by-pass must be screwed right down.If the cooker is to work on natural gas, pro-ceed as follows:-Remove the oven bottom (loosen the screw to remove the bottom);-ignite the oven burner, turning the knob pointer to the maximum setting;-shut the oven door;-access the thermostat or tap by-pass (see fig. 8);-back off the thermostat by-pass by about 3 turns;-after 5 or 6 minutes, turn the knob pointer to the minimum setting;-slowly re-tighten the by-pass, watching the flame decrease in size through the window in the closed oven door until the tongue of the flame is about 4 mm long. Never keep the flame too low. It must be stable even when the oven door is opened or closed quickly;-turn off the burner and replace the oven bottom.CONNECTING TO THE ELECTRICAL MAINSBefore making the connection, check that: -the mains voltage is as indicated on the nameplate;-the earth connection is in good working or-der.If the socket is not easily accessible, the in-stallation engineer must provide a switch with a contact breaking gap of 3 mm or more.If the appliance power lead is not fitted with a plug, use an approved standard type, re-membering that:-the green-yellow wire must be used for the earth connection;-the blue wire is the neutral;-the brown wire is live;-the lead must never touch hot surfaces overabout 75 degrees C;-replacement leads must be of type H05RR-F or H05V2V2-F of suitable size (see dia-grams in fig. 2).-if the appliance is supplied without lead,using type H05RR-F or H05V2V2-F cableof suitable size (see diagrams in fig. 2).IMPORTANT: the manufacturer declines all liability for damage due to failure to comply with the regulations and standards in force.Check that the appliance is correctly con-nected to the earth (see diagrams in fig. 2 at the back of the manual).THE SAFETY DEVICEThe correct gap between the end of the ther-mocouple sensor and the burner is shown in figures 5 and 6.To check that the valve is working properly, proceed as follows:-ignite the burner and leave it to work forabout 3 minutes;-turn off the burner by returning the knob tooff position (-turn the knob pointer to the "on" position;-release the knob in this position and movea burning match towards the burner; ITMUST NOT IGNITE.Time needed to excite the magnet during ignition: 10 seconds approx.Automatic tripping time, after flame has been turned off: not more than 60 seconds for oven and grill burners.IMPORTANT-Before doing any work inside the cooker, dis-connect the mains plug and shut the gas tap.-Never use matches to check the gas cir-cuit for leaks. If a specific control device isnot available, foam or very soapy water canbe used.-When re-closing the hob, check that theelectrical wires of the spark plugs (ifpresent) are not close to the injectors, sothat they cannot run across them.36HOW TO USE THE COOKER VENTILATIONAll gas cooking appliances produce heat and moisture in the rooms where they are in-stalled. Take care to ensure that the kitchen is well ventilated; keep the ventilation open-ings unobstructed or install an extractor hood with fan.In case of intensive or prolonged use, addi-tional ventilation may be required; open a window, or increase the extractor fan power. IGNITING THE HOB BURNERS-Press the knob and turn it anti-clockwise-at the same time, move a burning match towards the burner head;-to reduce the flame, turn the knob further in the same direction until its pointer issition).IGNITING THE OVEN BURNER-Open the oven door;-press the knob and turn it anti-clockwise to the maximum flame position;-move a burning match towards the hole in the centre of the oven bottom and press the knob right down (see fig. 9);-check that the burner has ignited, looking through the hole in the centre of the bot-tom, keeping the knob pressed all the time; -after about 10 seconds, release the knob and check that the burner remains on. Oth-erwise, repeat the operation.IGNITING THE GRILL BURNER (GAS GRILLS)-Fit the control knob guard as shown in fig. 12;-press the oven knob and turn it to the right until it reaches the stop;-move a burning match towards the perfo-rated burner pipe and press the knob right down (see fig. 10);-check that the burner has ignited, keepingthe knob pressed down;-after about 10 seconds, release the knoband check that the burner remains on. Oth-erwise, repeat the operation.IMPORTANT-Difficulty in igniting burners is normal if thecooker has been out of use for some time.The air accumulated in the pipes will beexpelled in a few seconds;-Never allow too much unburnt gas to flowfrom the burners. If ignition is not achievedwithin a relatively short time, repeat the pro-cedure after returning the knob to the offposition (-time, a smell may be noticed and smokemay come out of the oven. This is becauseof the surface treatment and oily residueson the burners.SAFETY DEVICEBurners equipped with this device have the advantage that they are protected if they accidentally go out. If this occurs, the supply of gas to the burner concerned is automati-cally cut off, preventing the hazards deriving from a leak of unburnt gas. The gas supply must be cut off within no more than 60 sec-onds for the oven and grill burners.HOW TO USE THE HOB BURNERSUse pans of diameter suitable for the burner type. The flames must not project beyond the base of the pan. Recommended sizes: -for auxiliary burners = pans of at least 8 cm -for semi-rapid burners = pans of at least 14 cm -for rapid burners = pans of at least 22 cm.N.B.: Never keep the knob at settings be-off position .FOR COOKERS EQUIPPED WITH ELEC-TRIC HOTPLATESThe different heat settings are obtained as follows:37- 1 = minimum setting for all hotplates;- 6 = maximum setting for normal and rapid hotplates (with red disc);- 0 = off.Pans must never be smaller in diameter than the hotplates and their bottoms must be as flat as possible (see fig. 11). IMPORTANT:-Never leave hotplates on without pans, except when first used; leave for about 10 minutes to dry oil or moisture residues;-if the hotplate is to be out of use for a long time, apply a little grease to its painted surface; -do not allow spills to burn onto the hotplate, requiring the use of abrasive cleaners. HOW TO USE THE GAS OVEN-After igniting the burner, leave the oven to heat up for about 10 minutes;-place the food for cooking in an ordinary oven dish and place it on the chrome-plated shelf; -place the food in the oven, using the shelf on the third pair of runners whenever possi-ble, and turn the knob pointer to the desired setting;-cooking can be observed through the window in the door with the oven light on. This will avoid opening and closing the door frequently, unless oil or fat has to be added to the dish. IMPORTANT: never place foods directly on the drip tray for cooking; it is there only to collect any drips of fat during grilling.N.B.: For cookers without thermostat:-270 degrees C-150 degrees C-All other temperatures between 150 and 270 degrees C are obtained approximately by positioning the knob between the maxi-mum and minimum settings.Never leave the knob in positions between theHOW TO USE THE GAS OR ELECTRIC GRILL-fit the knob guard (see fig. 12);-ignite the burner and wait a few minutes toallow it to warm up, or switch on the heat-ing element;-place the foods on the chrome-plated shelf;-insert on the highest runner;-insert the drip tray on the bottom runner;-gently close the oven door, resting it againstthe knob guard;-after a few minutes, turn the food to exposethe other side to the infrared radiation (thecooking time depends on the type of foodand personal taste).N.B.: the first time the grill is used smoke will come out of the oven. Before inserting foods for cooking, wait until any oil residues on the burner have completely burnt away.The grill must only be used at its full rated heat.IMPORTANT: accessible parts may be hot when the grill is in use! Keep children well away.The grill element in the top of the oven is switched on by turning the thermostat knob clockwise to the grill symbol on the control panel.The red light will come on to show the ele-ment is in operation.The table below will serve as a guide; bear-ing in mind that cooking times and tempera-tures may vary depending on the type and amount of foods cooked and personal taste.Food to be Time (minutes)grilled1st side2nd sideThin pieces of meat 6 4Fairly thick pieces ofmeat 8 5Thin fish or fishwithout scale10 8Fairly thick fish1512Sausages1210Toasted sandwiches 5 2Small poultry2015CONVENTIONAL ELECTRIC OVEN-The oven shelf is designed to take normaloven dishes for cooking sweets or roasts,3839-The oven shelf is designed to take normal oven dishes for cooking sweets or roasts,or is used without a pan for cooking foods under the grill.-The drip tray is only there to collect any juice from foods and must never be used as a cooking surface.OVENS WITH THERMOSTATIf cooking temperatures are not as set, call in an engineer to check the thermostat.WARMING COMPARTMENTTo open the warming compartment, open the flap door with one hand (see figure 13).To close the warming compartment, simply press the flap door back into place.GENERAL PRECAUTIONS-Always disconnect the power supply before any work inside the oven or where live parts may be accessed.-Never use the warming compartment for storing inflammable liquids or items which do not withstand heat, such as wood, pa-per, aerosol cans, matches, etc.-Make frequent checks on the rubber con-nection hose, ensuring that it is well away from hot surfaces, that there are no sharp bends or kinks, and that it is in good condi-tion. The hose must be replaced at the lat-est at the indicated date and must be se-cured at both ends using a standard hose clamp.-If taps become stiff to operate over time,contact the After-Sales service.-Wash enamelled or chrome-plated parts with soapy lukewarm water or non-abrasive detergents. A metal brush may be used to remove deposits from hob burners and flame caps. Dry thoroughly.-Never use abrasives to clean enamelled or chrome-plated parts.-Do not use too much water when washing the hob. Take care that no water or other substances enter the burner housing holes,as this may be dangerous.or is used without a pan for cooking foods under the grill.-The drip tray is only there to collect any juice from foods and must never be used as a cooking surface.There is a single control knob for the oven or grill.Starting from the 0 (off) position, the knob can be turned clockwise to the following settings:--The spark plugs for electric ignition mustbe kept clean and dry; always check after use, particularly if there have been drips or overflows from pans.-Never close glass lids until the hob burn-ers or hotplates have cooled completely; it might shatter or crack.-Never knock enamelled parts or ignition spark plugs (where present).-The main or wall gas tap should be turnedoff when the cooker is not in use.-Never lift the cooker by taking hold of the oven door handle.No liability is accepted for injury or damage caused by poor installation or improper useof the cooker.In case of malfunctions, particularly gas leaks or short-circuits, contact your en-gineer without delay.404142587634mm 2 - 4HBA43121011ED. 09.02.1999334813B1/44。
- 1、下载文档前请自行甄别文档内容的完整性，平台不提供额外的编辑、内容补充、找答案等附加服务。
- 2、"仅部分预览"的文档,不可在线预览部分如存在完整性等问题,可反馈申请退款(可完整预览的文档不适用该条件!)。
- 3、如文档侵犯您的权益，请联系客服反馈,我们会尽快为您处理(人工客服工作时间：9:00-18:30)。
体内自噬研究工具-AAV-mRFP-GFP-LC3关于自噬：自噬（autophagy）自噬是细胞在自噬相关基因(autophagy related gene，Atg)的调控下利用溶酶体降解自身受损的细胞器和大分子物质的过程。
自噬是生物机体的一种细胞内部压力协调与代谢机制，所以，自噬是发生在细胞内部的，以细胞为发生单位的一种表现。
关于自噬的作用讨论详见《汉恒生物-自噬（Autophagy）及其研究方法概述》技术文件。
Zhifen Yang&Daniel J.Klionsky,Nature Cell Biology12,814–822 (2010)细胞自噬自噬发生在细胞内部的一种机制，而千千万万的细胞形成了组织，器官，系统，最终协调形成了复杂的生命综合体。
正是由于自噬是细胞内部的一种压力应对机制，研究自噬时，除了LC3IIB的Western blot检测和电镜检测，我们经常还需要将细胞从实验动物中分离，并应用汉恒LC3双标腺病毒（Ad-mRFP-GFP-LC3：汉恒LC3双标，自噬研究金指标）进行体外细胞自噬流实时检测。
汉恒LC3双标-细胞自噬流检测体系：mRFP-GFP-LC3融合蛋白的病毒产品。
mRFP用于标记及追踪LC3，GFP的减弱可指示溶酶体与自噬小体的融合形成自噬溶酶体，即由于GFP荧光蛋白对酸性敏感，当自噬体与溶酶体融合后GFP荧光发生淬灭,此时只能检测到红色荧光。
通过显微镜成像后，计算红黄光点的数量以衡量自噬流的强弱；比较红黄点的数量，以评价自噬流的平衡状态。
体内自噬研究汉恒通过将体内基因转染利器-腺相关病毒红绿荧光merge后通过merge后出现的黄色斑点即只是自噬体.红色的斑点指示自噬溶酶体,通过不同颜色斑点的计数可以清晰的看出自噬流的强弱。
如下图:细胞转染mRFP-GFP-LC3病毒后给予氨基酸剥夺处理2小时后出现明显增强的自噬以及自噬流。
自噬是细胞内的一种“自食（Self-eating）”的现象，凋亡是“自杀（Self-killing）”的现象，二者共用相同的刺激因素和调节蛋白，但是诱发阈值和门槛不同，如何转换和协调目前还不清楚。
自噬是指膜（目前来源还有争议，大部分表现为双层膜，有时多层或单层）包裹部分胞质和细胞内需降解的细胞器、蛋白质等形成自噬体，最后与溶酶体融合形成自噬溶酶体，降解其所包裹的内容物，以实现细胞稳态和细胞器的更新。
目前文献对自噬过程进行观察和检测常用的策略和手段有：通过western blot检测LC3的剪切；通过电镜观测自噬体的形成；在荧光显微镜下采用GFP（-RFP）-LC3等融合蛋白来示踪自噬体形成以及降解。
近几年对自噬流的研究日趋增多，针对于此我们汉恒生物科技（上海）有限公司自主研发了用于实时监测自噬(流)的mRFP-GFP-LC3腺病毒，mRFP用于标记及追踪LC3，GFP的减弱可指示溶酶体与自噬小体的融合形成自噬溶酶体，即由于GFP荧光蛋白对酸性敏感，当自噬体与溶酶体融合后GFP荧光发生淬灭,此时只能检测到红色荧光。
这种串联的荧光蛋白表达载体系统直观清晰的指示了细胞自噬流的水平，是我们自噬研究尤其是自噬流研究不可或缺的利器。
一、腺相关病毒属微小病毒科(parvovirus)，为无包膜的单链线状DNA病毒。
AA V 的基因组约4700bp，包括上下游两个开放读码框架(ORF)，位于分别由145个核苷酸组成的2个反向末端重复序列(ITR)之间。
AAV基因组中有3个启动子(P5、P19和P40)和2个开放阅读读框(ORF)，rep和cap，如图1所示，它们在AA V病毒整合、复制和装配中其重要作用。
图1.AA V基因组结构二、腺相关病毒的优点1.安全性高：迄今从未发现野生型AA V对人体致病，重组AA V基因组序列上去除了大部分的野生型AA V基因组元件，进一步保证了安全性；进行操作，而且进行动物实验时造成的免疫反应小；3.宿主范围广：能感染分裂细胞和非分裂细胞；4.表达稳定：能介导基因的长期稳定表达；5.物理性质稳定：在60℃不能被灭活，能抗氯仿；三、CRISPR/Cas9基因敲除原理CRISPR(clustered,regularly interspaced,short palindromic repeats)是一种来自细菌降解入侵的病毒DNA或其他外源DNA的免疫机制。
II型CRISPR/Cas系统在发挥功能时仅需要一种蛋白即Cas9核酸酶参与。
Cas9核酸酶复合体由Cas9蛋白和两个非编码的RNA-crRNA前体pre-crRNA和tracrRNA组成。
在酿脓链球菌(Streptococcus pyoge nes)中,pre-crRNA和tracrRNA均由CRISPR位点上转录而来,然后pre-crRNA通过其含有的重复序列和tracrRNA形成RNA异二聚体,并结合到Cas9蛋白上,Cas9蛋白再对pre-crRNA进行剪切以获得成熟的crRNA,成熟的crRNA上长度为20个碱基的向导序列通过与目的DNA的序列互补来引导整个Cas9复合体去切割目的DNA。
除了成熟的crRNA上的向导序列外,在Cas9识别的DNA位点的3’端,还必须有一个PAM(protospacer adjacent motif)区域。
而Cas9识别位点的特异性就由20个碱基的向导序列和3个碱基的PAM序列共同决定。
一旦基因组序列中出现由于gRNA引导的Cas9切割DNA断口，细胞内的非同源末端连接修复机制和同源重组修复机制就会启动，而非同源末端连接修复机制会造成基因组序列的突变和移码，从而有一定概率出现目的基因的序列突变和蛋白翻译提前终止。
图2.Crispr/Cas9系统示意图四、pHBAAV-SaCas9-gRNA基因元件示意图SaCas9（staphylococcus aureus cas9）来自金黄色葡萄球菌，基因长度由经典的spCas9（Streptococcus pyogenes）的4400bp缩短为3300bp，基因大小大幅减少了25%。
SaCas9可以被装载进入腺相关病毒（AAV）而不影响AAV的包装和活力，因此提供了一个非常方便有效的在体基因操作工具。
图3.pHBAAV-SaCas9-gRNA示意图五、sgRNA设计及pHBAAV-SaCas9-gRNA载体构建1.以Human TRF3基因为例进行说明CDS区（灰色标记为第一外显子，黄色标记为第二外显子）：ATGGCCTCTGCGCCCTGGCCGGAGCGGGTTCCGAGGCTGCTCGCTCCGCGCTTACCCTCTTACCCGCCCCCACCCCCAACAGTGGGAT TACGGTCCATGGAGCAGGAGGAGACCTACCTGGAGCTCTACCTGGACCAGTGCGCCGCTCAGGATGGCCTTGCCCCACCCAGGTCTC CCCTGTTCAGCCCAGTTGTACCTTATGATATGTACATACTGAATGCATCCAATCCGGATACTGCATTTAATTCGAACCCTGAAGTCAAAG AAACATCTGGTGATTTCTCATCTGTGGATCTTAGCTTCCTACCAGATGAAGTTACCCAGGAAAATAAAGACCAGCCTGTCATTAGCAAA CACGAAACTGAAGAAAATTCTGAAAGCCAAAGTCCACAAAGTAGGTTGCCATCACCCAGCGAACAGGACGTTGGGCTGGGCTTAAA CAGCAGCAGTTTGTCAAATTCCCATTCACAGCTGCACCCTGGTGATACTGACTCAGTCCAGCCCTCTCCTGAGAAACCAAACTCCGAC TCCTTGTCTCTGGCATCCATAACTCCCATGACACCAATGACCCCTATTTCAGAATGTTGTGGAATTGTACCTCAACTACAGAATATAGTT TCCACTGTAAACCTGGCCTGTAAGTTGGATCTGAAGAAAATAGCTTTGCATGCAAAAAATGCAGAATATAACCCAAAGAGGTTTGCTG CTGTCATAATGAGGATCCGAGAGCCCAGGACAACAGCCCTTATATTTAGCTCTGGGAAGATGGTCTGCACGGGAGCCAAAAGTGAAG AGCAGTCTCGACTTGCAGCAAGAAAATATGCTCGTGTGGTGCAGAAGCTTGGGTTCCCTGCCAGATTCCTCGATTTTAAAATTCAGAA CATGGTTGGAAGCTGTGATGTGAGATTTCCCATCAGGCTGGAAGGTTTGGTGCTAACCCATCAGCAGTTCAGTAGTTACGAGCCTGAA CTGTTTCCTGGTCTTATTTATAGAATGGTAAAACCACGAATTGTGTTGCTTATCTTTGTATCTGGAAAAGTTGTGTTGACAGGTGCCAAA GAACGTTCTGAGATCTATGAAGCATTTGAAAACATCTATCCTATTCTAAAAGGTTTTAAAAAAGCCTGA2.SaCas9-gRNA设计原则：2.1SaCas9识别的PAM区为NNGRRT（NN GAAT，NNGAGT，NNGGAT，NNGGGT），NNGRRN也可以，对于sgRNA的长度，一般应为21或者22nt；2.2sgRNA序列的碱基组成，要避免序列中含“TTTT”终止信号，其中GC%含量最佳为40%~60%；2.3如果sgRNA是U6或T7启动子驱动表达的，要考虑sgRNA的5'碱基为G或GG，以提高其转录效率；2.4对于sgRNA靶向基因的结合位置，如需造成基因移码突变，需尽量靠近基因编码区的ATG下游，最好位于第一或第二外显子；2.5检查sgRNA靶向结合位点基因组序列是否存在SNPs；2.6对于Cas9单切口酶，设计paired-gRNA时需考虑成对sgRNA的间距；2.7全基因脱靶效应分析，需考虑脱靶位点最大允许几个错配碱基数，建议最少5个碱基。
重点考察种子序列和非种子序列碱基错配数，以及脱靶位点是否位于基因编码区等，另外还可考察是否存在碱基插入或缺失的脱靶位点。
3：SaCas9-gRNA序列设计：基于以上原则：最终确定的gRNA打靶序列为：SaCas9-gRNA1T:5’CACC G GACCCCTATTTCAGAATGTTG3’（正义链）SaCas9-gRNA1B:5’AAAC CAACATTCTGAAATAGGGGTCC3’SaCas9-gRNA2T:5’CACC GGTCATTGGTGTCATGGGAGTT3’（反义链）SaCas9-gRNA2B:5’3’SaCas9-gRNA3T:5’CACC GGCTGGACTGAGTCAGTATCAC3’（反义链）SaCas9-gRNA3B:5’4：pHBAAV-SaCas9-gRNA载体构建4.1将合成的配对gRNA序列退火成双链；4.2将PHBAAV-SaCas9-gRNA载体采用BsaI酶进行酶切和胶回收；4.3将退火的sgRNA与酶切好的PHBAAV-SaCas9-gRNA载体用T4连接酶连接；4.4测序确认sgRNA序列插入正确。