Zeiss LSM 510 META, Operating Manual

Page 1: ...LSM 510 and LSM 510 META Laser Scanning Microscopes Operating Manual Release 3 2...

Page 2: ...plication of this document and the utilization and communication of its contents are not permitted Violations will entail an obligation to pay compensation All rights reserved in the event of granting...

Page 3: ...will help you to find out which settings and methods are capable of improving or accelerating a procedure The HOT SURFACE symbol warns against hazards for the user that might arise when touching the l...

Page 4: ...977 Configuration 3 000000 1219 978 Configuration 4 000000 1219 979 Configuration 5 000000 1219 980 Configuration 9 000000 1219 981 Configuration 11 000000 1219 982 Configuration 12 000000 1219 983 C...

Page 5: ...em table 453031 0000 000 Small system table 453032 0000 000 System baseplate 000000 1171 342 Active anti vibration table 30 x 30 000000 1177 841 Active anti vibration table 1200 mm x 1400 mm 000000 11...

Page 6: ...checked above was installed and handed to the customer in functional condition on by Phone Fax The customer has been instructed on how to operate and maintain the equipment Place date Carl Zeiss Jena...

Page 7: ...In the Operation section you will find the most important steps and procedures of the LSM menu structure The step by step description how to get an image will be shown by typical application examples...

Page 8: ...INTRODUCTION LSM 510 Carl Zeiss LSM 510 META VIII B 45 0008 e 10 02...

Page 9: ...Y CONTENTS Page 1 NOTES ON DEVICE SAFETY 1 3 1 1 General 1 3 1 2 Regulations 1 3 1 3 Notes on Setting up the Microscope System 1 4 1 4 Notes on Handling the Computer and Data Media 1 5 1 5 Notes on Ca...

Page 10: ...NOTES ON DEVICE SAFETY LSM 510 Carl Zeiss Contents LSM 510 META 1 2 B 45 0008 e 10 02...

Page 11: ...se operating instructions and all device publications belonging to the system conscientiously before operating the LSM 510 You can obtain additional information on the hardware configuration delivered...

Page 12: ...efore connecting the mains cables please check whether your mains voltage corresponds to the voltage specified on the rating plate of the laser module For reasons of laser safety the TV port on the mi...

Page 13: ...dling diskettes avoid data losses by protecting them against extreme temperatures moisture and magnetic fields The data on a diskette is stored in the form of magnetic signals To some extent monitors...

Page 14: ...ncy or by persons authorized and trained for this purpose by the manufacturer Damaged units or parts may only be repaired or maintained by the responsible service agency Care operations that may be ca...

Page 15: ...aff The dangerous laser radiation area is limited to the beam path and to a distance of up to around 10 cm from the specimen Nevertheless you should observe the following warnings If necessary insofar...

Page 16: ...all of the labels shown below are provided on your instrument and contact Carl Zeiss Germany or one of the service agencies if you should discover that any of the labels should be missing You will rec...

Page 17: ...A NOTES ON DEVICE SAFETY LSM 510 Warning and Information Labels Carl Zeiss B 45 0008 e 10 02 1 9 Fig 1 2 Warning and information labels on the Axioplan 2 imaging MOT microscope with LSM 510 META scann...

Page 18: ...NOTES ON DEVICE SAFETY LSM 510 Carl Zeiss Warning and Information Labels LSM 510 META 1 10 B 45 0008 e 10 02 Fig 1 3 Warning and information labels on laser components page 1...

Page 19: ...ing and Information Labels Carl Zeiss B 45 0008 e 10 02 1 11 Fig 1 3 Warning and information labels on laser components page 2 COHEREnT EnTERPRISE E4 J3 7 SE AR CH PE AK J6 E4 E5 wa ter OU T max im um...

Page 20: ...NOTES ON DEVICE SAFETY LSM 510 Carl Zeiss LSM 510 META 1 12 B 45 0008 e 10 02...

Page 21: ...irements 2 11 2 6 Vibrations 2 12 2 7 Laser Specifications 2 12 2 7 1 Coherent Enterprise 653 II 352 364 nm 80 mW Laser Class 3 B 2 12 2 7 2 Point Source i flex 2000 405 nm 25 mW Laser Class 3 B 2 12...

Page 22: ...LSM 510 SETUP REQUIREMENTS LSM 510 Carl Zeiss Contents LSM 510 META 2 2 B 45 0008 e 10 02...

Page 23: ...croscope large system table 300 250 cm The system rack contains the laser module Helium Neon laser 543 nm 633 nm and Argon ion laser the power supply for the Argon ion laser for HBO lamp and halogen l...

Page 24: ...M with Ar UV Laser We recommend placing the cooling unit of the Ar laser UV in a separate room to prevent heat accumulation and vibration Length of the water hose 400 cm One microscope System Rack I 5...

Page 25: ...43 nm 633 nm and Argon ion laser and the Argon UV laser module The system rack II contains the power supplies for lasers for HBO and halogen lamps the electronic control unit ECU and the MCU28 unit if...

Page 26: ...d Stand Upright Stand also possible Fig 2 4 2 1 3 2 Spectra Physics MaiTai Direct coupling with Upright Stand Upright Stand also possible Fig 2 5 System Rack 500 2000 4000 1000 1400 Large NLO system t...

Page 27: ...tand also possible Fig 2 6 2 1 3 4 Coherent Chameleon Direct coupling with Upright Stand Inverted Stand also possible Fig 2 7 System Rack 500 2500 3500 1000 1400 Large NLO system table 1800 Power Plug...

Page 28: ...VA per phase Power plug CEE red 230 V 16 A 3 phases N PE phases 1 and 2 connected CEE yellow 115 V 32 A 3 phases N PE phases 1 and 2 connected Argon UV laser Line Voltage 208 240 V AC 10 50 60 Hz 208...

Page 29: ...ser Module VIS Fig 2 8 2 2 1 Phase 1 LSM feeds the following units Laser Module HeNe 2 via Power 1 5 socket adapter Computer monitor Microscope MCU28 Scanning Module via Power 2 HAL lamp HBO lamp 2 2...

Page 30: ...er or Chameleon 120 140 75 200 Active anti vibration table NLO For Mira or Tsunami Laser 180 140 75 400 Scanning Module LSM 510 25 20 25 15 Scanning Module LSM 510 META 28 27 30 5 13 Microscope 50 35...

Page 31: ...60 LSM 190 85 120 350 Monitor computer 120 80 90 80 UV laser unit 125 55 50 100 UV cooling unit 120 60 90 50 META scan head 52 47 47 13 META upgrade kit 64 5 60 5 42 5 20 2 5 Environmental Requirement...

Page 32: ...Specifications 2 7 1 Coherent Enterprise 653 II 352 364 nm 80 mW Laser Class 3 B Line voltage 208 240 V Line frequency 50 60 Hz Max current 1 phase at 208 V 34 Amps 230 V 31 Amps 240 V 29 Amps Power c...

Page 33: ...N 7812 458 477 488 514 nm 30 mW Laser Class 3 B Line voltage 100 240 V with factory setting Line frequency 50 60 Hz Max current 1 phases at 25 A Power consumption 2000 VA Cooling fan on top of laser h...

Page 34: ...objectives and accessories can be accommodated Z motor DC servomotor opto electronically coded Least Z interval 50 nm Axioplan 2 imaging MOT Axiovert 200 M BP or SP 100 nm Axioskop 2 FS MOT HRZ 200 G...

Page 35: ...ive Zoom 1 40 continuous control Channels a Up to 4 channels simultaneously or b 3 traditional confocal channels and 1 META channel 4 confocal reflection fluorescence channels PMT or 3 PMT and 1 META...

Page 36: ...405 nm 25 mW Ar laser 458 477 488 514 nm 30 mW ArKr laser 488 568 nm 30 mW Fuses and automatic circuit breakers for 230 V G type fuse 5 20 mm slow blow 3 15 A H 250 V acc to IEC 127 2 circuit breakers...

Page 37: ...LSM 510 LSM 510 SETUP REQUIREMENTS LSM 510 META System Overview LSM 510 META Carl Zeiss B 45 0008 e 10 02 2 17 2 12 System Overview LSM 510 META...

Page 38: ...LSM 510 SETUP REQUIREMENTS LSM 510 Carl Zeiss System Overview LSM 510 META LSM 510 META 2 18 B 45 0008 e 10 02...

Page 39: ...LSM 510 LSM 510 SETUP REQUIREMENTS LSM 510 META System Overview LSM 510 META NLO Carl Zeiss B 45 0008 e 10 02 2 19 2 13 System Overview LSM 510 META NLO...

Page 40: ...LSM 510 SETUP REQUIREMENTS LSM 510 Carl Zeiss System Overview LSM 510 META NLO LSM 510 META 2 20 B 45 0008 e 10 02...

Page 41: ...ODUCTION TO Laser Scanning Microscopy 3 3 3 1 Principle of Laser Scanning Microscopy 3 3 3 2 Three Dimensional Presentations of LSM Image Stacks 3 4 3 3 Optical Diagram of the LSM 510 Schematic 3 6 3...

Page 42: ...INTRODUCTION TO LASER SCANNING MICROSCOPY LSM 510 Carl Zeiss Contents LSM 510 META 3 2 B 45 0008 e 10 02...

Page 43: ...nce of fluorescent specimens would disturb the in focus image of object point of interest resulting in a blurred image of poor contrast The problem therefore is how to capture only the light coming im...

Page 44: ...ods to extract spatial information from stacks of slice images Gallery The simplest presentation of 3D information is a gallery showing the individual slice images sections of a stack arranged side by...

Page 45: ...ually perpendicular sectional images Oblique sections A section through the stack is made along an oblique plane defined by the selection of five coordinates i e X Y Z angle of rotation and angle of t...

Page 46: ...m Splitter NDF Neutral Density Filter VP Variable Pinhole PMT Photomultiplier T PMT Transmission Photomultiplier The diagram above is a schematic representation of the LSM system Laser light is focuse...

Page 47: ...ion channels usable for reflection or fluorescence and an additional transmitted light channel are ideal for the investigation of multiple fluorescence specimens Separately in each of the four channel...

Page 48: ...ed objective nosepiece 5 H DIC is firmly fixed to the stand where no operating elements can be found for the nosepiece Operation will be performed via LSM 5 software control The Restriction of the nos...

Page 49: ...utter preventing laser light from reaching the stand when the support is moved to the back A complementary shutter built in the stand prevents laser light from reaching the eyepieces during the scanni...

Page 50: ...itting fast change over between methods as well as automatic operation Parameters once set or complex examination sequences once established can be saved and reproduced therefore complete application...

Page 51: ...4 2 1 Log on to WINDOWS NT 4 8 4 2 2 Switching on the Enterprise UV Laser 4 10 4 2 3 Starting the LSM 5 Program 4 10 4 3 Quick Start in the Expert Mode 4 13 4 3 1 Start the Expert Mode 4 13 4 3 2 Set...

Page 52: ...QUICKSTART LSM 510 Carl Zeiss Contents LSM 510 META 4 2 B 45 0008 e 10 02...

Page 53: ...graphic 32 bit Microsoft WINDOWS NT 4 0 operating system and WINDOWS 2000 respectively Portions Copyright 1996 Microsoft Corporation All rights reserved The installation of the software for the LSM 51...

Page 54: ...el Limited function range within a window List box or selection box Selection of one of the displayed options at a click of the mouse Open the box by clicking on the arrow button Input box Input of te...

Page 55: ...tes deactivates setting options Button Selection performance of a function via mouse click 4 1 3 Convention for the Text in this Manual All the originally used terms of the software interface e g name...

Page 56: ...ter krn Pinhole setting files AIM PH pos Log files AIM log The following files generated during the system integration should also be included in a backup procedure Parameter file for pinhole setting...

Page 57: ...started If REMOTE CONTROL switch is not used turn the system on with the I button on the laser module in addition the jumper plug supplied must be connected to the POWER REMOTE CONTROL terminal Turn t...

Page 58: ...being loaded The Begin Logon window appears on the screen 4 2 1 Log on to WINDOWS NT Press the three keys Ctrl Alt and Del at the same time The Logon Information window appears on the screen permitti...

Page 59: ...er entries confirm by clicking the OK button or Enter The WINDOWS NT operating system desktop appears on the screen showing a number of icons Fig 4 5 WINDOWS NT operating system desktop Change Filters...

Page 60: ...e the entire program package image recording and analysis is available while only a part of the software functions image analysis only of already stored images and no hardware functions are available...

Page 61: ...ou to process and analyze previously acquired images with the LSM 5 software In this mode control of the hardware laser module is not possible off line mode Please note that the Scan New Images button...

Page 62: ...itialization is performed and can be monitored in the Initialization window and interrupted with a click on the Cancel button if required Depending on the selected option Scan New Images or Use existi...

Page 63: ...and channel assignment tracks multi tracks switching on off of laser lines and intensity excitation image creation determine the scan method line frame and scan parameters image size scan speed pixel...

Page 64: ...Convallaria specimen You can view the specimen in either fluorescence reflected light or transmitted light To view the specimen in transmitted light set the Reflector Turret position to None and acti...

Page 65: ...ht panel by clicking on the Transmitted Light button Select the On button and control the intensity by the slider Select an objective with low magnification by clicking in the Objective panel of the A...

Page 66: ...ng the appropriate wavelength to excite the dyes used to label your specimen In the case of the argon and UV laser Enterprise click on the Standby button first Warming Up appears in the line Status Af...

Page 67: ...configurations Tracks are available in the Track Configurations window which can be used for fast and easy image acquisition The list of configurations will appear by clicking on the button Choose the...

Page 68: ...screen The microscope must be in the LSM mode press LSM button and move the relevant slider s on the microscope stand to the LSM position On the right hand side of the Scan Control window various but...

Page 69: ...2 labels FITC Rhod with defined channels is easier to view in split screen where each channel is arranged side by side You can toggle between xy and Split xy in the Image Display window The scanned im...

Page 70: ...still allow the Detector Gain setting and to provide sufficient image information 1 Airy unit is a decent value to obtain a confocal XY image use the 1 button If required adjust the pinhole again see...

Page 71: ...ld improve signal noise dramatically however the image acquisition rate will be slower When the image optimization has been finished click on the Single button to generate a single image of the specim...

Page 72: ...of the Image Display window The Save Image and Parameter As window appears on the screen Proceed as follows to store images in an existing database To select the required database click on its name b...

Page 73: ...s you like Before clicking on the Create button in the Create New Database window set the location in which the database will be created by selecting the drive in the Create in list box and double cli...

Page 74: ...ivate Laser off or Exit in the Shutdown tab The lasers will then automatically be switched off when you exit the LSM program 4 4 1 Exiting the LSM Program Close all open windows of the LSM program by...

Page 75: ...0008 e 10 02 4 25 4 4 2 Shut Down the WINDOWS Operating System Move the cursor to the bottom margin of the screen This opens the taskbar containing the Start button Click on the Start button of the ta...

Page 76: ...fter WINDOWS NT has been run down the Shutdown Computer window appears which tells you that you can now turn off your computer 4 4 3 Turning Power Off Please bear in mind that a cooling phase of at le...