Zeiss Lattice Lightsheet 7 Instructions Manual
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Zeiss Lattice Lightsheet 7 Instructions Manual

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April 2021

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  • Page 1 April 2021...
  • Page 2 Violations will entail an obligation to pay compensation. All rights reserved in the event of granting of patents or registration as a utility patent. Issued by: Carl Zeiss Microscopy GmbH Carl-Zeiss-Promenade 10 07745 Jena, Germany microscopy@zeiss.com www.zeiss.com/microscopy...

  • Page 3: Table Of Contents

    CONTENTS Lattice Lightsheet 7 ZEISS CONTENTS Page SAFETY INFORMATION .................. 5 General ..........................5 Explanation of Symbols ......................6 Information Concerning the Use of the Device ..............7 Regulations ........................... 8 Power Requirements ......................9 Environmental Requirements ....................14 Notes on Setting up the System ..................15 Warning and Information Labels ..................
  • Page 4 Components of Lattice Lightsheet 7 module ................ 64 2.10 System Overview ......................... 65 OPERATING INSTRUCTIONS ................. 66 Switching on and off the Lattice Lightsheet 7 System ............67 3.1.1 Starting and Stopping the ZEN Software ................68 3.1.2 Presetting the System ......................70 3.1.2.1...

  • Page 5: Safety Information

    ZEISS 1 SAFETY INFORMATION General Lattice Lightsheet 7 is a stand-alone light sheet system in an inverted configuration. It consists of the Lattice Lightsheet system proper, laser cassette, incubation components, camera with liquid cooling system and computers. Lattice Lightsheet 7 systems are designed for research only and not for medical diagnostic, therapeutic or treatment purposes.

  • Page 6: Explanation Of Symbols

    WINDOWS operating system and its graphical user interface. The respective manuals are supplied together with the programs. The Lattice Lightsheet 7 system are laser hazard class 2 devices. The systems are equipped with safety interlocks that comply with laser hazard class 3B. WHO recommendations concerning health and industrial protection when handling laser devices must be observed.

  • Page 7: Information Concerning The Use Of The Device

    Information Concerning the Use of the Device The Lattice Lightsheet 7 systems meet the EMC requirements for EN 55011 Class A (intended use in industrial environment). If the system is operated in a residential area or in a small trade area other devices may be influenced by conducted or radiated disturbance.

  • Page 8: Regulations

    2012/19/EC (WEEE), if applicable for the product. ZEISS has installed a process for taking back and recycling the instruments within the member states of the European Union, which takes care of the appropriate utilization according to the said EU guidelines.

  • Page 9: Power Requirements

    The following components must be connected to the power supply unit (PSU): - laser module (LM) - Lattice Lightsheet 7 module - 6-point power strip (multipoint connector 1 or system components power strip) The storage and data analysis PC and respective monitor must be separately connected to the AC network by further power cables.
  • Page 10 700 W max. 700 W max. The Lattice Lightsheet 7 system must be connected to the AC network via the Power Supply Unit (PSU) by a country-specific mains power cable (Fig. 1). The 6 point power strip must be separately connected to the AC network by further power cables.
  • Page 11 SAFETY INFORMATION Lattice Lightsheet 7 Power Requirements ZEISS Fig. 1 Schematic drawing of power connections 04/2021 V_01 000000-2324-960...
  • Page 12 5 TRIGGER STAGE 14 TEMP (Supply line) 6 TRIGGER CAM-1 15 TEMP (CAN Bus) 7 TRIGGER OUT 16 MICO-F-1 8 TRIGGER CAM-2 17 POWER CONNECTOR 9 LVDS Fig. 2 Connectors and fuses of the Lattice Lightsheet 7 000000-2324-960 04/2021 V_01...
  • Page 13 Fig. 3 Component rack with power supply unit (PSU), laser module (LM), and motion controller (MC) for Lattice Lightsheet 7 (rear panels) The PSU and the LM are pre-mounted, the MC will be mounted on-site. The mains plug may only be inserted into a socket with a protective contact. The protective effect must not be overridden by an extension cable without a protective conductor.

  • Page 14: Environmental Requirements

    For the same reason, keep the cardboard packaging of other major components, such as the Lattice Lightsheet 7 module and user PC sealed during acclimation. For operation the system must be placed in a closed room.

  • Page 15: Notes On Setting Up The System

    The Lattice Lightsheet 7 systems are delivered in several crates. The Lattice Lightsheet 7 system must be set up so as to ensure that the minimum clearance between the wall and the rear of the system is no less than 0.5 m. This clearance is needed for adjustment and maintenance operations.
  • Page 16 By establishing a corresponding workplace environment, please ensure that the formation of electrostatic charges is avoided. To avoid vibrations during operation, the Lattice Lightsheet 7 systems should only be operated in conjunction with the system table (vibration damping). The Lattice Lightsheet 7 should be placed in a room suitable and dedicated for microscopes.

  • Page 17: Warning And Information Labels

    ZEISS Warning and Information Labels The warning and information labels attached on the Lattice Lightsheet 7 system (Fig. 4 and Fig. 5) must be observed. Check whether all of the labels shown below are provided on your instrument, and contact ZEISS Germany or one of the service agencies if you should discover that any of the labels should be missing.
  • Page 18 SAFETY INFORMATION ZEISS Warning and Information Labels Lattice Lightsheet 7 Laser ON LED US version only Fig. 5 Warning and information labels of a Lattice Lightsheet 7 system (front view top, rear view bottom) 000000-2324-960 04/2021 V_01...

  • Page 19: Notes On Handling The Laser Components And Illumination Systems

    Caution: Use of controls or adjustments or performance of procedures other than those specified herein may result in hazardous radiation exposure. The following laser types are currently intended for use in the Lattice Lightsheet 7 system. The use of any other lasers as the ones listed below is not authorized.

  • Page 20: Laser Lifetimes

    4 mW 1.11 Notes on Handling the Computer, Data Media and Data Transfer The computer used as standard in your Lattice Lightsheet 7 system is a high-end computer with WINDOWS operating system. As standard, your computer has one hard disk drive and one CD-ROM drive.

  • Page 21: Notes On Care, Maintenance And Service

    The ZEN software is released with a specific windows version and patch and this must not be changed unless approved by Carl Zeiss Microscopy. Please contact your ZEISS service representative to check the safety of the device. Please keep the service logs of your device.

  • Page 22: Replacing Fuses And Electrical Connections

    SAFETY INFORMATION ZEISS Notes on Care, Maintenance and Service Lattice Lightsheet 7 1.12.2 Replacing Fuses and Electrical Connections Please use only original parts for replacing fuses or power cables. In the event of an error in the mains power supply of individual components or the whole system you can change the mains fuses 10 A.

  • Page 23: Maintaining The Liquid Cooling System (Lcs-Bu) Of The Camera

    1.12.5 Count on Service in the True Sense of the Word Because the ZEISS microscope system is one of your most important tools, we make sure it is always ready to perform. What’s more, we’ll see to it that you are employing all the options that get the best from your microscope.

  • Page 24: User Interface - Mounting And Dismounting Components

    All interface ports which can potentially emit laser radiation are equipped with special TORX safety screws which must not be loosened by the user. The following interfaces are no user interfaces when used with a Lattice Lightsheet 7. - pco.edge sCMOS cameras and their adapters...

  • Page 25: Changing The Meniscus Lens (Relay Optics)

    SAFETY INFORMATION Lattice Lightsheet 7 User Interface - Mounting and Dismounting Components ZEISS 1.13.1 Changing the Meniscus Lens (Relay Optics) The meniscus lens (relay optics) is mounted on a metal support. Meniscus lens plus support must be exchanged as a whole.
  • Page 26 SAFETY INFORMATION ZEISS User Interface - Mounting and Dismounting Components Lattice Lightsheet 7 3. Remove the incubator lid (Fig. 9/3), see also section 1.13.3. 4. Remove the sample mounting frame (Fig. 9/4), see also section 0. 5. Remove the meniscus lens holder (Fig. 9/5).
  • Page 27 SAFETY INFORMATION Lattice Lightsheet 7 User Interface - Mounting and Dismounting Components ZEISS 8. Release the Luer-Lock couplings (Fig. 11/9 and /10). 9. Remove the old meniscus lens. 10. Place the new meniscus lens in the meniscus lens holder. 11. Connect the Luer-Lock couplings.

  • Page 28: Removing The Immersion Supply System

    SAFETY INFORMATION ZEISS User Interface - Mounting and Dismounting Components Lattice Lightsheet 7 1.13.2 Removing the Immersion Supply System 1.13.2.1 Removing the Immersion Container There is an immersion container for water supply. In order to refill or empty a container press the front door (Fig.

  • Page 29: Immersion Priming

    SAFETY INFORMATION Lattice Lightsheet 7 User Interface - Mounting and Dismounting Components ZEISS 1.13.2.2 Immersion Priming The immersion supply system is depicted in Fig. 14. It is a water immersion system, so only water should be used as immersion. Do not use oil- or glycerol-based immersion media, as these more viscous solution will clog the tubing system (Fig.
  • Page 30 ZEISS User Interface - Mounting and Dismounting Components Lattice Lightsheet 7 The first immersion before mounting the sample must be supplied manually. During this priming process air- bubbles will be removed from the immersion system. Proceed as follows (Fig. 14): 1.

  • Page 31: Mounting And Dismounting The Incubator Lid

    SAFETY INFORMATION Lattice Lightsheet 7 User Interface - Mounting and Dismounting Components ZEISS 1.13.3 Mounting and Dismounting the Incubator Lid The incubation system is an integral part of the system. The lid is attached to the stage by magnets. Before mounting a sample, the incubator lid must be removed.

  • Page 32: Exchanging The Sample Mounting Frames

    SAFETY INFORMATION ZEISS User Interface - Mounting and Dismounting Components Lattice Lightsheet 7 1.13.4 Exchanging the Sample Mounting Frames The stage proper with the incubation unit can receive sample mounting frames with different formats. Dismounting: 1. Remove the incubator lid.

  • Page 33: Supplementing Liquid To The Liquid Cooling System

    Read the operating manual of the liquid cooling system carefully. Note that the liquid cooling system is connected via its ground plug to the laser module of the Lattice Lightsheet 7 by a red cable to equalize potential differences. 04/2021 V_01...

  • Page 34: Incubation System

    ZEISS User Interface - Mounting and Dismounting Components Lattice Lightsheet 7 1.13.6 Incubation System The Incubation of the Lattice Lightsheet 7 consists of 2 systems: The Heating System comprising: · Temperature Controller to monitor temperature · Heated lid · Heated plate The Gas Incubation System comprising: ·...
  • Page 35 Connect channel 1 / heated lid, channel 2 / heated tray, and channel 3 / heated base (Fig. 19/9, 10, 11) with the gas supply plug line plug (Fig. 2/12) of the Lattice Lightsheet 7. There is no need to connect the spare (heated lens) plug (Fig. 19/12).
  • Page 36 SAFETY INFORMATION ZEISS User Interface - Mounting and Dismounting Components Lattice Lightsheet 7 Heated Lid The heated lid (Fig. 20) is fixed to the heated incubation chamber via magnets. It consists of the following components: · Metallic frame · Electronic contacts ·...

  • Page 37: Gas Incubation System

    It is intended to be used with stage top incubator of the Lattice Lightsheet 7. The gas incubation system mixes the required gas ratios in the output and actively humidifies this mixed gas output before the gas enters the incubation chamber. The system can control the concentration and humidity.
  • Page 38 SAFETY INFORMATION ZEISS User Interface - Mounting and Dismounting Components Lattice Lightsheet 7 Hydraulic Connections The gases from the bottle or laboratory gas line (for Air/O also a compressor can be used) need to be connected to the Gas Mixer with the OD 6 mm polyurethane tubing of the respective color. This tubing is delivered with the system.
  • Page 39 SAFETY INFORMATION Lattice Lightsheet 7 User Interface - Mounting and Dismounting Components ZEISS Humidifying Column Tubing connector red (output) 5 Red label: Water level min. 50 ml Tubing connector blue (input) 6 Red label: Water level max. 130 ml Electric plug...
  • Page 40 SAFETY INFORMATION ZEISS User Interface - Mounting and Dismounting Components Lattice Lightsheet 7 Hydraulical connections 1. Connect the blue marked tubing side from the Heated Tubing Package with the blue marked Tubing connector (input) (Fig. 23/2). The other white marked end of this tubing is connected to the white labeled gas mixer output 1 (Fig.
  • Page 41 SAFETY INFORMATION Lattice Lightsheet 7 User Interface - Mounting and Dismounting Components ZEISS Heated Tubing Package The Heated Tubing Package (Fig. 24) connects the gas output of the Gas Mixer to the Humidifying Column and to the Heated Chamber on the microscope stage. The gas tubing carrying humidified gas is heated up to avoid condensation.

  • Page 42: Operation

    Lattice Lightsheet 7 system (Fig. 2/11). The CAN connector TEMP socket of the Lattice Lightsheet 7 (Fig. 2/14) remains free. 3. Plug the electric cable from the Lattice Lightsheet 7 system (Fig. 21/8) to electric socket of the chamber frame docking block (Fig. 21/2).
  • Page 43 11. Connect channels 1-3 of the Temperature Controller (Fig. 25/9a, b, c) to the TEMP supply line socket of Lattice Lightsheet 7 (Fig. 25/9e). Do not connect channel 4. In addition, connect the electrical plug at the front of the Lattice Lightsheet 7 to the electrical socket of the stage top incubator (Fig. 25/2a).
  • Page 44 13. Connect the CAN bus of the Gas Mixer to the Temperature Controller (Fig. 25/8c) and the Lattice Lightsheet 7 (Fig. 25/8b). Fig. 25 Electric and gas circuits of the heating and gas incubation systems 1 Lattice Lightsheet 7 2 Heated chamber Gas connection tube Electrical connection cable...
  • Page 45 SAFETY INFORMATION Lattice Lightsheet 7 User Interface - Mounting and Dismounting Components ZEISS 7 Gas supply source supply source Air supply source supply source 8 Gas Mixer Supply line CAN bus CAN bus 9 Temperature controller Supply line Supply line...

  • Page 46: Sw Control Of Incubation

    SAFETY INFORMATION ZEISS User Interface - Mounting and Dismounting Components Lattice Lightsheet 7 1.13.6.4 SW Control of Incubation The Heating System and the Gas Mixer System are controlled via the ZEN SW in the Incubation tab (Fig. 26). Alternatively, ibidi’s Incu Control SW can be used.

  • Page 47: Temperature Adjustment Set

    SAFETY INFORMATION Lattice Lightsheet 7 User Interface - Mounting and Dismounting Components ZEISS 1.13.6.5 Temperature adjustment set The temperature for Lid, Base and Tray may need to be set at higher values than the temperature to be used in the experiment. Required setting values can be tested with the Temperature adjustment set.

  • Page 48: Stage Control

    User Interface - Mounting and Dismounting Components Lattice Lightsheet 7 1.13.7 Stage Control 1.13.7.1 Stage Motion Controller The stage of the Lattice Lightsheet 7 is controlled by the Motion controller (Fig. 30). It can move on 5 axes: · X · Y · Z ·...
  • Page 49 SAFETY INFORMATION Lattice Lightsheet 7 User Interface - Mounting and Dismounting Components ZEISS For further information please consult the "Five Axes Microscope Stage S119458000-XX User Guide" from Nanomotion Ltd. Debug Connector 9 Input / Output channel 1 analog input/output pins, for factory use...

  • Page 50: Ergo Pointer

    SAFETY INFORMATION ZEISS User Interface - Mounting and Dismounting Components Lattice Lightsheet 7 1.13.7.2 Ergo pointer The Ergo pointer can be used to control the stage remotely (Fig. 31). It can move the stage in X (Fig. 31/6), Y (Fig. 31/7) and Z (Fig. 31/2). It is connected via its USB port (Fig.

  • Page 51: Detection Module (Pco.edge)

    A pco.edge 4.2 CLHS sCMOS camera is attached to this camera port (Fig. 32). A cooling liquid defined as a hazardous substance is used in the Lattice Lightsheet 7 to cool the detection modules (depending on configuration). The supplied safety data sheet with notes on hazards and safety measures must be observed when handling the cooling liquid.

  • Page 52: Cable Connections For The Detection Module

    SAFETY INFORMATION ZEISS User Interface - Mounting and Dismounting Components Lattice Lightsheet 7 1.13.8.1 Cable Connections for the Detection Module The detection module pco.edge 4.2 CLHS is equipped with a liquid cooling system (LCS). Please read the safety notes in chapter 1.13.5, as well as the separate safety data sheet for the liquid cooling system. The LCS is connected to the system's 6-point power strip.
  • Page 53 · The trigger cable is always connected to the detection module with [1/IN] and on the rear side of the main system module Lattice Lightsheet 7 to the Camera 1/IN socket (Fig. 32/1). · The detection module is switched on with the toggle switch (Fig. 32/10) on the module itself.

  • Page 54: Setup Requirements

    Lattice Lightsheet 7 2 SETUP REQUIREMENTS Space Requirements Lattice Lightsheet 7 systems on small system table Fig. 34 Lattice Lightsheet 7systems with system table and PC table (dim. in mm) The laser module and the power supply unit are housed in the component rack (Fig. 38). The stage controller as well as gas, humidity and temperature controller need extra space to be set up near the system.
  • Page 55 Lattice Lightsheet 7 system footprint on small system table (dim. in mm) The laser module and power supply unit for Lattice Lightsheet 7 can be housed inside the components rack, which should be positioned beside the microscope stand on the floor.

  • Page 56: Power Requirements

    The following components must be connected to the power supply unit (PSU): - laser module (LM) - Lattice Lightsheet 7 module - 6-point power strip (multipoint connector 1 or system components power strip) The storage and data analysis PC and respective monitor must be separately connected to the AC network by further power cables.
  • Page 57 700 W max. 700 W max. The Lattice Lightsheet 7 system must be connected to the AC network via the Power Supply Unit (PSU) by a country-specific mains power cable (Fig. 36). The 6 point power strip must be separately connected to the AC network by further power cables.
  • Page 58 SETUP REQUIREMENTS ZEISS Power Requirements Lattice Lightsheet 7 Fig. 36 Schematic drawing of power connections 000000-2324-960 04/2021 V_01...
  • Page 59 12 GAS (Supply line) 5 TRIGGER CAM-1 13 TEMP (CAN Bus) 6 TRIGGER OUT 14 TEMP (Supply line) 7 TRIGGER CAM-2 15 MICO-IF-1 8 LVDS 16 POWER CONNECTOR Fig. 37 Connectors and fuses of the Lattice Lightsheet 7 04/2021 V_01 000000-2324-960...
  • Page 60 Fig. 38 Component rack with power supply unit (PSU), laser module (LM), and motion controller (MC) for Lattice Lightsheet 7 (rear panels) The PSU and the LM are pre-mounted, the MC will be mounted on-site. The mains plug may only be inserted into a socket with a protective contact. The protective effect must not be overridden by an extension cable without a protective conductor.

  • Page 61: Physical Dimensions Of Items

    Physical Dimensions of Items ZEISS Physical Dimensions of Items Item Dimension L x W x H [cm] Weight [kg] Lattice Lightsheet 7 Module 60 x 42,5 x 38 48.0 Stage 40 x 34 x 9.5 Stage Motion Controller (MC) 36 x 32.5 x 9.5...

  • Page 62: Physical Dimensions Of Item Packages

    Lattice Lightsheet 7 Physical Dimensions of Item Packages Item Dimension L x W x H [cm] Weight [kg] Lattice Lightsheet 7 Meniscus Lens 25.8 x 25.8 x 16 Lattice Lightsheet 7 Cables 37 x 30 x 35 Stage Sample Holder Frame Inserts...

  • Page 63: Environmental Requirements

    9. Heat emission 0.8 kW Vibrations According to ISO 10811 operation of the Lattice Lightsheet 7 system is recommended for vibration class C (VC-C, 12.5 µm/s for 8-80 Hz (RMS)). Laser for Lattice Lightsheet 7 Single mode polarization preserving fiber Laser beam attenuation for all lasers by modulation ·...

  • Page 64: Components Of Lattice Lightsheet 7 Module

    3 Incubator 4 Slide-in module for immersion 5 Immersion container 6 Syringe 7 Meniscus lens holder 8 Front door 9 LLS 7 module 10 Camera 1 module Fig. 39 Components of Lattice Lightsheet 7 (LLS 7) system 000000-2324-960 04/2021 V_01...

  • Page 65: System Overview

    SETUP REQUIREMENTS Lattice Lightsheet 7 System Overview ZEISS 2.10 System Overview Fig. 40 System overview of the Lattice Lightsheet 7 system 04/2021 V_01 000000-2324-960...

  • Page 66: Operating Instructions

    3 OPERATING INSTRUCTIONS This instruction describes the basic operation of the Lattice Lightsheet 7 systems with the ZEN software. The purpose of this document is to guide the user to get started with the system as quick as possible to obtain some first images from his samples.

  • Page 67: Switching On And Off The Lattice Lightsheet 7 System

    Switching on and off the Lattice Lightsheet 7 System ZEISS 3.1 Switching on and off the Lattice Lightsheet 7 System The Lattice Lightsheet 7 system is switched on by the switches of the PSU (Fig. 41) Switching on the System 1. Switch on the system Power (Fig. 39/1).

  • Page 68: Starting And Stopping The Zen Software

    Starting and Stopping the ZEN Software Starting the ZEN Software · Double click the ZEN icon on your WINDOWS desktop to start the ZEISS ZEN software. The ZEN Starting Application window (Fig. 42) followed by ZEN Login window are displayed (Fig. 43).
  • Page 69 OPERATING INSTRUCTIONS Lattice Lightsheet 7 Switching on and off the Lattice Lightsheet 7 System ZEISS During the initialization phase the Stage/Focus not calibrated window (Fig. 46) might appear depending on the settings of the microscope. You can either skip the calibration by pressing the Skip calibration button or perform the calibration by pressing the Calibrate Now button.

  • Page 70: Presetting The System

    OPERATING INSTRUCTIONS ZEISS Switching on and off the Lattice Lightsheet 7 System Lattice Lightsheet 7 3.1.2 Presetting the System 3.1.2.1 Switching to Operation Mode (Obligatory) After initialization, the ZEN Lattice Lightsheet GUI in startup mode is displayed. The Imaging Setup tab (Fig.

  • Page 71: Calibration Of The Stage

    OPERATING INSTRUCTIONS Lattice Lightsheet 7 Switching on and off the Lattice Lightsheet 7 System ZEISS 3.1.2.3 Calibration of the Stage In order to calibrate the stage, press the Calibration Home XYZ button of the Stage LLS tab in the right tool area (Fig.

  • Page 72: Environmental Control (Optional)

    OPERATING INSTRUCTIONS ZEISS Switching on and off the Lattice Lightsheet 7 System Lattice Lightsheet 7 3.1.2.5 Environmental Control (Optional) Set the required temperature in the Temperature expansion field of the Incubation tab (Fig. 51) of the right tool area. You can select different temperatures for Lid, Tray and Base.

  • Page 73: System Operation

    OPERATING INSTRUCTIONS Lattice Lightsheet 7 System Operation ZEISS System Operation 3.2.1 Control Tabs Imaging Setup Track Selection Track Addition LED illumination (Transmitted Light) Stage with sample (fixed) Laser illumination (Epifluorescence) Illumination objective lens (fixed) Detection objective lens (fixed) Filter wheel Camera pco.edge 4.2 CLHS (fixed)
  • Page 74 OPERATING INSTRUCTIONS ZEISS System Operation Lattice Lightsheet 7 Acquisition Mode Scan definition Scan display Range in µm covered in y Number of slices per stack Step size (Interval) Light sheet selection Dither on/off switch Camera region of interest (ROI) in µm and pixel (px) Fig.
  • Page 75 OPERATING INSTRUCTIONS Lattice Lightsheet 7 System Operation ZEISS Auto Immersion Time interval setting Immersion supply mode Fig. 55 Auto Immersion tab Lattice Lightsheet / Aberration Control Movement of sheet along optical axis of illumination objective lens axis Movement of sheet perpendicular to illumination...

  • Page 76: Sample Positioning And Finding

    OPERATING INSTRUCTIONS ZEISS System Operation Lattice Lightsheet 7 3.2.2 Sample Positioning and Finding 1. Switch on the white LED lamp and set it to 20- 50%. (Do not use side illumination as you will lose contrast (Fig. 58). 2. Use None position in the filter wheel.

  • Page 77: Lightsheet Alignment And Stage Levelling

    OPERATING INSTRUCTIONS Lattice Lightsheet 7 System Operation ZEISS 3.2.3 Lightsheet Alignment and Stage Levelling Only necessary, if new sample/sample carrier has been inserted. 1. Switch TL LED off. 2. Switch on one laser light, e.g. 488 nm. 3. Put filter to “None” position.

  • Page 78: Lightsheet Focusing

    OPERATING INSTRUCTIONS ZEISS System Operation Lattice Lightsheet 7 3.2.4 Lightsheet Focusing 1. Switch of TL LED. 2. Choose appropriate laser line e.g. 488 and set to approximately 2% Fig. 62 Lightsheet tilt correction: uncorrected (upper panel) and X-tilt 3. Choose 1000 x 300 px corrected (lower panel) 4.
  • Page 79 OPERATING INSTRUCTIONS Lattice Lightsheet 7 System Operation ZEISS 13. Zoom on some structures in the sample and use aberration control (= Alvarez plates) to maximize sharpness & contrast. 14. Optionally save these settings for later use. Once the system is aligned only the stage is moved! Lightsheet and detection objective are fixed –...

  • Page 80: Sample Imaging

    OPERATING INSTRUCTIONS ZEISS System Operation Lattice Lightsheet 7 3.2.5 Sample Imaging Setup volume scan in Acquisition Mode (Fig. 67). Either use Center mode or First/Last Set the beam path as required. mode. Center Mode: 1. Press Center to center the stage at the current position.

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