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Etaluma LS850 Operator's Manual

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LS850 Microscope Manual 2025
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  • Page 1 For assistance, please email support@etaluma.com Etaluma,Inc. Carlsbad, California www.etaluma.com LS850 Microscope Manual 2025...

  • Page 2: Table Of Contents

    CONTENTS CONTENTS Contents 1 Introduction 1.1 Included Items ..........1.2 Accessories: .
  • Page 3 11 Appendix B: Setting Up Time-Lapse In Windows 10 11.1 Power Settings ..........11.2 Connectivity Settings .

  • Page 4: Introduction

    1 Introduction 1.1 Included Items Items included with each LS850 Microscope: • Auraphase transmitted illumination • LED fixture with cable • Optional ambient light guard • Fixture mounting knob Figure 1: Items Included with LS850 LS850 Microscope Manual 2025...

  • Page 5: Accessories

    1.2 Accessories: • USB A & USB C communication cables • Dichroic cleaning swabs • Black microplate lid for blocking room light during fluorescence imaging • Foam plug for incubator port • Lens paper • USB memory stick with LumaViewPro application and this manual •...

  • Page 6: Labware Holders

    1.3 Labware Holders Primary Labware Holder Figure 3: Primary Holder Optional Labware Holders • Four microscope slides in parallel plastic • Four microscope slides in parallel metal with magnetic clamps Figure 4: Optional Holders LS850 Microscope Manual 2025...

  • Page 7: Setting Up The Microscope

    2 Setting Up the Microscope 2.1 Connecting the Cables Connect the USB type B end to the LS850 at the rear left corner of the microscope. Wait to connect the type A or C USB cable end from the Microscope to the computer until LumaViewPro is installed.

  • Page 8: Auraphase Transmitted Illumination Accessory

    2.2 AuraPhase Transmitted Illumination Accessory Remove the AuraPhase from its shipping box. Attach it by placing the holes on the bottom of the bracket over the pegs on the upper surface. Tighten the large thumb screw beneath the deck cup. Connect the free end of the AuraPhase Accessory power cable to the round socket nearby.

  • Page 9: Setting Up Lumaviewpro

    3.1 Introduction The LS850 is controlled by the LumaViewPro software program. The latest LumaViewPro version is downloadable from Etaluma’s website and should be installed prior to connecting your computer for the first time. LumaViewPro can be run on Windows10/11, Mac, and Linux. Desktop computers and laptops can be used, but the best visualization correlates with monitor resolution equal to the sensor resolution (up to 2100 x 2100 pixels).

  • Page 10: Connecting The Ls850 Microscope

    3.3 Connecting the LS850 Microscope Insert the USB-A end(some versions may be C) of the supplied USB cable into a USB3 port on your computer and the other USB-B end into the port labelled “Computer” on the microscope. It is also important to connect the Microscope directly to the computer USB port and not use a USB hub.

  • Page 11: Troubleshooting

    4 Troubleshooting 4.1 Microscope Not Connected This communication error will occur under the following circumstances: 1. No power connected, check AC source and cables. 2. USB cable disconnected, check connections on PC and microscope. 3. USB cable connected to USB2 port on the PC, switch to USB3 port. Figure 7: Communication Error Screen LS850 Microscope Manual 2025...
  • Page 12 (a) USB2 Connection (b) USB3 Connection Figure 8: USB Connection Scenarios The Device Manager entry for the microscope plugged into a USB2 port is shown in the first dropdown while the second shows the proper connection to a USB3 port. LS850 Microscope Manual 2025...

  • Page 13: Menus Inaccessible

    4.2 Menus Inaccessible Certain computers and their monitors may need the Display Scale changed to a lower percent- age. The default (Recommended) Scale can cause some menus to have their lower portions hidden and inaccessible. Changing this to a lower percentage will allow the full menu to be visible.
  • Page 14 Figure 11: Settings>System>Display>Scale LS850 Microscope Manual 2025...

  • Page 15: Getting Started With Lumaviewpro

    5 Getting Started With LumaViewPro 5.1 Launch Launch the LumaViewPro software from the desktop icon (or other chosen location) and allow the microscope to be discovered and initialize. Launching LumaViewPro opens the application as seen below: Figure 12: Launch Screen LS850 Microscope Manual 2025...
  • Page 16 The side menus are accessed by clicking the blue arrows on the right and left edge of the application. Figure 13: Left: Motion Control Menu Right: Channel Control Menu LS850 Microscope Manual 2025...

  • Page 17: Microscope Settings

    5.2 Microscope Settings Figure 15: Objective Settings Figure 14: Microscope Settings Click the Objective field to select the desired Click on the left arrow to expand the motion control objective from the dropdown. menus and select the Microscope Settings tab. Microscope Settings configures your hardware with camera, objective, and image settings.
  • Page 18 Enable Full Pixel Depth saves all ac- Video Recording Format controls video quired images as 12-bit tiffs (default is output for manual and protocol video 8-bit). capture. If mp4 is selected, videos will be saved as a playable mp4 video in Live Image Format allows simple tiff real-time FPS.
  • Page 19 Live Image Histogram Equalization en- hances the contrast of the live image, ex- panding the histogram to be able to bet- ter visualize small differences in contrast; dark pixels become darker, bright pixels become brighter. The underlying image data is not changed, only the displayed images are modified.

  • Page 20: Channel Control

    5.3 Channel Control Expanding the right side menu exposes the Channel Control. There are three transmitted channels- Brightfield, Phase Contrast, and Extended Phase. Brightfield illumination is from a central LED with an appropriate lens for uniform illumination. Phase Contrast is a small ring of LEDs that match the IPC objectives we supply from Evident.

  • Page 21: Xy Stage Control

    5.4 XY Stage Control The location of the stage will be indicated by a red crosshair in the Stage Map field. Clicking within the Stage Map will move to that location. Arrow keys allow coarse and fine XY move- ments according to the objective installed. Setting and returning to XY positions is controlled here.

  • Page 22: Objective Control

    5.5 Objective Control Adjust Focus The Focus control allows manual adjustment of the focus position using coarse and fine arrows according to the objective installed. You may drag the objective icon up or down for very large movements. Clicking within the objective icon will adjust focus based on the position relative to the center of mass of the icon.
  • Page 23 Turret Selection If configured as an 850T, the Turret position can be selected. The focus will retract before changing positions and will need to be brought back into focus through the “Go to Set Z” button or manual adjustment. When using only two objectives, they should be installed in opposite locations.

  • Page 24: Getting An Image

    6 Getting an Image 6.1 Initial Set Up Place the sample in the nest and ensure it is seated flat. Click the Stage Map to move to the location of your sample or first well of your microplate. Enable an illumination channel and adjust the focus to find the image.

  • Page 25: Protocol

    6.3 Protocol To configure an automated microscopy run, including time-lapse, the Protocol feature is uti- lized. Time lapse Protocols are created when the New button is clicked. The following settings are captured upon creation: • Channels which have their Acquire option checked will be used with their corresponding Image/Video, Illumina- tion, Gain, and Exposure settings as well as their, False color, Saved Focus, Auto Gain/Exp, and Autofocus en-...
  • Page 26 Tiling Adjacent images can be acquired in patterns of 2x2 up to 10x10. Tiling dimension is captured at Protocol creation and cannot be Inserted or Changed after creation. Tiling names are auto- generated and are required for Stitching in Post Processing. Tiling with smaller frame sizes will improve the alignment of the abutted tile images.

  • Page 27: Configuring A Protocol Quick Start

    6.4 Configuring a Protocol Quick Start • Place sample into plate nest • Move to an example FOV by clicking on the plate map under Protocol or XY Stage Control • Adjust image parameters to center the pixel histogram (get a greyscale background) •...

  • Page 28: Creating And Editing Custom Labware

    7 Creating and Editing Custom Labware 7.1 Labware File Location The labware.json file-Labware definitions are stored in the labware.json file within the data folder. Figure 27: JSON File In the Data Folder The labware.json file can be opened with Notepad or equivalent text editor. 7.2 Adding a labware entry Each labware entry is defined by the information contained between the two “},”...
  • Page 29 Figure 28: Editing the JSON File In a Text Editor LS850 Microscope Manual 2025...

  • Page 30: Xy Stage Control

    7.3 XY Stage Control The XY Stage Control in LumaViewPro will display the X and Y coordinates and these values can be used for measuring the offset and spacing mentioned below. X is across the plate rows and Y is the up and down the columns. Diameter - This parameter is in millimeters.
  • Page 31 Figure 30: Diameter Adjustment LS850 Microscope Manual 2025...
  • Page 32 Figure 31: Spacing Adjustment LS850 Microscope Manual 2025...
  • Page 33 (a) Editing Columns and Rows (b) Editing XY Offsets Figure 32: Column and Rows & Offset Adjustment LS850 Microscope Manual 2025...

  • Page 34: Quickly Deleting Wells Including Tiles And Z-Stacks From A Protocol

    7.4 Quickly Deleting Wells Including Tiles and Z-Stacks From a Protocol Often when using tiling, a large numbers of labware wells may not be used and this represents many unwanted steps. These steps can be deleted in the Protocol menu on a step by step basis but in those cases where a large number of steps need to be deleted, one can edit the Protocol file to remove these wells quickly.

  • Page 35: Live Cell Imaging With The Ls850 Microscope In An Incubator

    8 Live Cell Imaging with the LS850 Microscope in an Incubator Prior to imaging within an incubator, perform the following steps where appropriate: 8.1 Installing Room Temperature LumaScope into a Warm Humid Incubator If a room temperature LumaScope is placed into a warm humid incubator, condensation will form on the microscope.

  • Page 36: Post Processing

    9 Post Processing 9.1 Video Creation Create Video allows time lapse or video frame recorded im- ages to be made into a movie. Under Post Processing se- lect Create Video. Apply Video Gen to Protocol Folder allows selection of the image files to be made into the AVI. The images will not be visible when selecting the folder.
  • Page 37 Stitch allows tiled images to be combined into a montage. Stitching is applied to a Protocol data folder and all tiles with the same well number, channel and sequence number are auto- matically stitched into a new image file (figure 36). Composite Generation will composite the fluorescence channel images that were acquired at the same location and in the same interval sequence.

  • Page 38: Object Analysis

    9.2 Object Analysis The Object Analysis capability allows simple thresh- olded objects to be filtered by Area and Perimeter as well as roundness and intensity. To analyze a folder of images, start by loading an example image and entering the Pixel Ratio appropriate for the objective magnification according to the following table: Figure 37: Object Analysis Menu The plot feature allows the visualization of the distribution of object intensity vs.
  • Page 39 Clicking Apply Method to Preview will outline the objects that meet the criteria. Threshold is generally between 5% and 30% as higher or lower thresholds will tend to select the entire image or none of it. Once settings have been optimized, the Method can be saved and then applied to a Folder of images.
  • Page 40 Figure 39: Method Preview Figure 40: Results Table LS850 Microscope Manual 2025...

  • Page 41: Z-Projections

    9.3 Z-Projections When a 3D dataset has been gathered as a stack of 2D images, different Z-Projection meth- ods can provide insight by applying various mathematical operations to the dataset to help examine different aspects of the data. Maximum Intensity Displays the highest intensity value from all slices at each (X,Y) position.

  • Page 42: Appendix A: Sanitizing The Microscope

    Further, it reaches all incubator walls, shelves, reservoirs, air plenums, sensors, and other interior components, and it leaves only small amounts of sterile water droplets as a residual. For questions, contact Technical Support at support@etaluma.com. LS850 Microscope Manual 2025...

  • Page 43: Appendix B: Setting Up Time-Lapse In Windows 10

    11 Appendix B: Setting Up Time-Lapse In Windows 10 11.1 Power Settings 1. Navigate to Settings (click on the Windows icon on the lower left of your screen and then the gear icon). Choose System, then Power & Sleep. On the right side column click on Additional Power Settings.
  • Page 44 (a) Advanced Power Settings (b) Sleep Settings. (c) USB Power Settings (d) Display Settings Figure 41: Customizing Advanced Power Settings LS850 Microscope Manual 2025...

  • Page 45: Connectivity Settings

    11.2 Connectivity Settings A.) Network connection NOT required (recommended configuration): 1. Disconnect the Ethernet cable (if wired connection is used) and turn on Airplane Mode. 2. Navigate to Settings -> Network & Internet -> Airplane Mode (left side) -> ON. Airplane Mode will prevent all wireless communications (Wi-Fi, Bluetooth and cellular) between the computer and other computers and devices.

  • Page 46: Appendix C: Setting Up Time-Lapse In Windows

    12 Appendix C: Setting up Time-Lapse in Windows 11 12.1 Power Settings Navigate to Settings (click on the Windows icon on the lower left of your screen and then the gear icon). Choose System, then Power & Battery. Set all to Never.

  • Page 47: Windows Update Settings

    12.3 Windows Update Settings 1. Navigate to Settings -> Windows Update -> Advanced Options (Fig 43) 2. Under “Choose how updates are installed” -> uncheck the checkbox next to “Enabling this policy will automatically download updates. . . ”. 3. Next, Click on Delivery Optimization and choose “Allow Downloads from other PCs”...

  • Page 48: Appendix D: Extending The Distance From Lumascope To Pc Workstation

    13 Appendix D: Extending the Distance from LumaScope to PC Work- station Often the LumaScope is located in an incubator which does not allow the PC or laptop to be placed in a convenient place for operation within the 3 meters that the included USB cable permits.

  • Page 49: Appendix E: Configuring Autogain

    14 Appendix E: Configuring Autogain Autogain brightness and timeout are stored in a json file which can be edited in Notepad and similar text editors. You can find these files under the data folder within the LumaViewPro folder. In the data folder you will find the settings.json file prior to the first use and then the current.json file after that.
  • Page 50 Figure 45: Power Settings For Range Extension Figure 46: File Location For Adjusting Autogain Brightness LS850 Microscope Manual 2025...
  • Page 51 Figure 47: Editing Autogain Settings LS850 Microscope Manual 2025...

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