Etaluma LS620 Manual de usuario

LS620, LS560, LS460 Microscopes

Operator’s Manual

With

Etaluma’s Lumaview Software

For assistance, please call 760-298-2355 or email support@etaluma.com.

Etaluma, Inc.

Carlsbad, California

www.etaluma.com

Lumaview™ is the trademark of Etaluma, Inc.

LS560/LS460 (with Phase Contrast

Accessory and Manual XY Stage, both

optional and ordered separately)

LS620 (with Phase Contrast Accessory

and Manual XY Stage, both optional

and ordered separately)

LS Manual Microscope Startup Manual 2021.5.1

Table of Contents

Setting Up the LS Microscope

A. Items Included With Each LS Microscope

B. Optional Accessories (purchased separately)

C. Recommended Computer Specifications

D. Connecting the Cables

E. Phase Contrast

F. Brightfield

G. Installing Objectives

H. About Lumaview

I. Downloading and Installing Lumaview

J. Connecting the LS Microscope

II.

Software: Getting Started with Lumaview

A. Starting Conditions

B. Launching Lumaview

C. Instrument Setup

D. Finding a Live Image

E. Focusing

F. Snapping Images

G. Auto-Generation of Single Composite Images

H. Live Video

I. Writing and Using Protocols

J. Run Protocol

K. Placing the Lumascope Microscope in an Incubator

L. Creating Videos

III.

Appendices

A. Windows Configuration and Driver Conflicts Troubleshooting

B. How to Disinfect Lumascopes for Incubator Placement

C. Setting up Windows10 for Uninterrupted Time-Lapse

LS Manual Microscope Startup Manual 2021.5.1

This document is available for download at https://etaluma.com/products/downloads.

Any steps specific for a particular LS model are noted in italics.

SETTING UP THE LUMASCOPE MICROSCOPE

Items Included With Each LS Microscope

1. LS620 and LS560

•Brightfield Illuminator (Unless you ordered Phase Accessory)

•USB power/communication cable

•Fluorescence shroud (black microplate lid)

•External power supply/cord with country-specific plug

2. LS460

•Brightfield

•USB power/communication cable

Optional Accessories (purchased separately)

•Phase Contrast Accessory (Olympus)

•Manual XY Stage

•Labware Holders

■

Holder for 35 mm Petri dishes, fits inside Holder for 60 mm Petri dishes

■

Holder for 60 mm Petri dishes & Terasaki plates, SBS outer dimensions

■

Holder for microscope slides & 50 mm Petri dishes, SBS outer dimensions

■

Holder for 4 microscope slides in parallel, SBS outer dimensions

•Light Hood

•Lumaquant, image analysis software

•Microvolution, deconvolution software

Recommended Computer Specifications

All LS Microscopes with Lumaquant: Windows 10; Core i7 or better processor; 500 GB to 1 TB SSD

hard drive, 4 Gb minimum RAM(8 Gb min. Lumaquant); single 4K monitor or two with 1080p HD

resolution. Windows must have Service Pack 2.0 with .NET version 4.5.2.

Microvolution deconvolution plugin for ImageJ/Fiji: A GPU is required; the most powerful NVidia

chip running CUDA that can be afforded is recommended for the best performance.

Connecting the Cables

1. Wait to connect the USB cable from the LS Microscope to the computer until Lumaview is

installed.

2. (LS620 and LS560 only). For almost all fluorescence imaging applications, power from the

computer via the USB cable to the LS Microscope will be sufficient. If higher levels of

illumination are required,e.g., with very dim samples or for photo bleaching(FRAP), insert the

LS Manual Microscope Startup Manual 2021.5.1

small round plug of the external power supply/cord into the round port labeled

SUPPLEMENTAL POWER on the back of the LS Microscope and the electrical plug into an AC

outlet. You should see an increase in the Illumination level beyond the 40% limit as indicated

on the Illumination slider in Lumaview with USB only.

Phase Contrast

1. If using the Phase Contrast Accessory, ensure that the Phase Accessory adaptor is attached to the

rear tab of the LS Microscope Deck. Attach the Phase Accessory by inserting the base of the arm

into the Phase Accessory adaptor. Ensure it is fully seated and tighten the thumb screw on the

bottom of the adaptor.

Phase accessory adaptor

Phase setup attached to Lumascope

2. Connect the free end of the Phase Accessory communication cable to the round port on the LS

Microscope back panel (left side) labeled External Lamp Power Out. This allows the Phase

Accessory to be controlled by Lumaview.

3. Insert the phase contrast Slider (pre-centered version included with Phase Accessory) into

the slot going through the condenser. Of the 3 positions in the Slider, the phase ring is

present in the middle position. A short lever on the front of the phase condenser controls the

iris. The iris should be fully open (to the left) for phase contrast.

LS Manual Microscope Startup Manual 2021.5.1

Brightfield

1. A flexible LED light source is provided if no Phase accessory is purchased. This

illumination allows direct and many oblique illumination setups.

2. For optimal brightfield, use the Phase Accessory with the included Slider in an

open position (no phase ring) and the iris closed down (fully right). The Phase

Accessory, which provides collimated light, is directly connected to the LS

Microscope, allowing it to be controlled by Lumaview. Transmitted light images

will be in gray scale due to the monochromatic CMOS camera.

3. Ambient light in a typically lit area can often be used. The light should be as

uniform as possible to prevent shadows or other dark or light areas. Overhead

fluorescent lighting can cause ballast interference -- a striped pattern on the

image is observed.Turn off the fluorescent lights or move the Lumascope to

another room.

Installing Objectives

1. Remove the triangular insert in the deck. Unscrew the black cap (or previously

installed objective) on top of the inside optics block and screw in the new objective.

Do not overtighten. Return the removable insert to the deck.

3. When installing or changing an objective, carry out the insertion quickly to

minimize dust falling onto the mirrored dichroic filter. A small amount of dust

will not affect imaging. However, if dust is significant and it appears to affect

quality of the live image or snapped images, contact Etaluma by email

(support@etaluma.com) to discuss possible methods for cleaning.

IMPORTANT: Using compressed air can result in an oily liquid being sprayed

onto the dichroic, which may damage it.

About Lumaview

1. LS Microscopes are controlled by the Lumaview software program. The latest

Lumaview version is downloadable from Etaluma’s website and must be

installed prior to connecting your computer for the first time to the LS

Microscope.

LS Manual Microscope Startup Manual 2021.5.1

2. Lumaview runs on Windows 10. Both desktop computers and laptops can be

used, but the best visualization correlates with monitor resolution equal to the

sensor resolution (up to 1900 x 1900 pixels for the LS620).

3. Note: The monitor does not affect image resolution unless the monitor is of such

low quality that it affects your ability to judge focus.

The computer should have Windows .NET Framework 4.5.2 or higher installed;

see https://msdn.microsoft.com/en-us/library/bb822049%28v=vs.110%29.aspx

for more information. If your computer does not have .NET Framework 4.5.2 or

higher installed, downloading Lumaview may automatically take you to the

Microsoft .NET download page. Scroll down the page to find the correct download.

This will not require a purchase. Contact Etaluma with any questions.

Downloading and Installing Lumaview

1. To download Lumaview, go to https://etaluma.com/products/downloads (under

the Resources tab). Click on Lumaview - ZIP link to start the download and save

the folder when prompted. Go to your Downloads folder and click to extract, then

open the Lumaview.zip file.

Alternatively, the same Lumaview.zip file can be copied from the flash drive that

comes with the Lumascope Microscope. Verify that this file is the latest version as

posted on the Etaluma website.

2. If your computer has an older version of Lumaview, installing the newest Lumaview

will overwrite the older version but retain all user data and information.

If there is a need to revert to an older version Lumaview, it will be necessary to

first uninstall the current version through Windows.

3. To install Lumaview, right click on the .zip file and Open with Windows Explorer.

Double click on the .msi installer file to start the installation. If a Windows

warning box about an unrecognized App appears, click More… then Run anyway.

You will be asked about the location; note that the default is a new Etaluma

folder inside the Program Files (x86) folder.

During installation, a Device Driver Installation Wizard will open. Click to

continue and finish (twice) installing the two drivers. Installation of

Lumaview will then finish. After installation, a Lumaview shortcut (orange circle

icon) convenient for launching the software will be present on the desktop.

Connecting the LS Microscope

1. If connecting with a computer that has been off, turn the computer on. Make sure the

computer is connected to its monitor(s). Insert the standard USB-A end of the

supplied USB cable into a USB port on your computer and the other square USB-B

end into the square port labeled TO COMPUTER on the back of the LS620, LS560 and

LS460. It is important to connect the LS Microscope directly to the computer USB

port and not use a USB hub.

LS Manual Microscope Startup Manual 2021.5.1

2. If Lumaview has not been run previously, it is important that the LS Microscope and

computer be connected using the USB cable before launching Lumaview. This is

because Windows needs to load the USB drivers before Lumaview can run. After the

first time, the program can be launched before connecting the LS Microscope and

computer. In the latter case, only the Outer Main Window will open. Connecting the

LS Microscope and computer with the USB cable will result in opening of the Live

Image Window as well.

II. SOFTWARE: GETTING STARTED WITH LUMAVIEW

The following sections give instructions for getting started with Lumaview on the LS620, LS560 and

LS460 and are intended as a Startup Manual only. Not all options are given and not all features may

be explained in any particular section. The complete User Guide for Lumaview is contained in the

Help Section of Lumaview. Click on Help in the Title Bar to open the pull down menu and click

Contents, or simply Click F1 to open the Help Section. Becoming familiar with the complete Help

Section is a must for using LS Microscopes optimally and taking advantage of the many features of

these Microscopes and accompanying software.

Starting Conditions

1. Lumaview has been downloaded and installed on the computer.

2. The LS Microscope is connected to the computer via the UBS cable.

3. Sample to be imaged is present on the LS Microscope stage and objective is installed.

4. LS620 or LS560: Fluorescence shroud (black microplate lid) is over sample if imaging in

fluorescence.

Launching Lumaview

Launch the Lumaview software from the desktop icon (or other chosen location) and allow

the LS Microscope to be discovered. This may take many seconds the first time.

Lumaview is installed with the Configuration set to a Lumascope model LS720 by default.

The following ERROR message will appear indicating that the software has not found the

LS Manual Microscope Startup Manual 2021.5.1

automated LS720 model and thus this will need to be changed to your respective model in

the Configuration>Instrument Settings Menu seen below. Click OK, change the model

selection and restart Lumaview.

Two windows will open on your monitor: Live Image Window inside the larger Main

Window. “Live Image Window” will be in the title bar when the Window is not maximized.

When Live Image Window is maximized, [Live Image Window] transfers to the end of the

Main Window title bar. The Live Image Window status bar at the bottom shows

communication with the camera sensor via continuous display of several useful metrics:

current frame rate, data transfer rate, number of frames collected thus far in the session,

frame size, and session date and time start.

Note: A variety of Windows configuration errors can

be displayed at this point. Please consult Appendix A.

LS Manual Microscope Startup Manual 2021.5.1

The Live Image Windows includes a Left Toolbar with the following commands:

Select Channel, optimize settings

Snap Image

Composite

Video Record (toggles on and off)

Protocol

Zoom

Shortcut to Last Image file location

When Lumaview is open, the icon that resides in your task bar appears as a document with an

arrow.

Instrument Setup

1. Click on Configuration in the File Menu bar to

open the drop-down menu and select

Instrument Setup to open its dialog box.

2. Check the Lumaview version to make sure it

is the most recently posted version on the

Etaluma website

(www.etaluma.com/products/downloads/).

Ensure that the LS Microscope being used is

checked. If it is necessary to change the LS

Microscope model number that is checked,

you will be prompted to restart the program.

Be sure to restart so that all functions work

correctly.

3. Confirm the current model is selected.

4. Click the Objective button to open its dialog box

(called Objective Lens Selection). Select the desired

objective to highlight it. Note that other fields about

the objective and image automatically fill based on

magnification and a frame size of 1200 x 1200 pixels.

Click OK. Once selected, the same information

automatically fills in Instrument Setup.

LS Manual Microscope Startup Manual 2021.5.1

5. Click the Frame Size button to open its dialog box. Using the pull-down menu, select

frame size desired (frame size will be the number times itself because it is square).

Note: If larger Frame Sizes are used, there will be some darkening in the corners.

If faster frame rates are needed, use smaller frame sizes, e.g., 800 x 800 pixels.)

If the Frame Size is changed, there will be a prompt to

restart Lumaview. Click OK to close the prompt and then

shut down Lumaview. Upon restarting Lumaview, the

frame size in the lower status bar will be the new size. If not

restarted, the new frame size may show in Instrument Setup

but it may not actually be different. Always check the lower

left status bar of Live Image Window to see the actual frame

size.

6. Continue to review and adjust the various settings in Instrument Setup. For Root,

enter desired file name root for all images in this imaging session. Select the file format

desired (Tiff is recommended).

7. The Time-lapse folder path default is given but can be changed if desired. Time-lapse

is set up under Protocol as described below. Note that the folder name Lumaview720

is the current default name and is used for images snapped on the other Lumascope

Microscopes as well unless a different folder is chosen.

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