Etaluma LS720 Manual de instalación y operación

IMPORTANT: This Manual describes how to set up LS Microscopes (Section I) and

start using Lu aview software (Section II). It is i portant that you follow this Guide.

Steps described for LS620 also apply to LS600.

For assistance, please call 760-298-2355 or e ail support@etalu a.co .

LS620 (with Phase Contrast Accessory

and Manual XY Stage, both optional

and ordered separately)

LS560/LS460 (with Phase Contrast

Accessory and Manual XY Stage, both

optional and ordered separately)

LS720 (with Phase Contrast

Accessory, optional and ordered

separately)

Etalu a, Inc.

www.etalu a.co

Lu aview™ is the trade ark of Etalu a, Inc.

LS720, LS620, LS600, LS560, LS460 Microscopes

Startup Manual

with

Etalu a’s Lu aview Software

LS Microscope Startup Manual 2018.7.25

Table of Contents

I. Setting Up the LS Microscope

A. Ite s Included With Each LS Microscope

B. Optional Accessories (purchased separately)

C. Reco ended Co puter Specifications

D. Placing the LS Microscope

E. Re oving the Shipping Lock (LS720 only)

F. Connecting the Cables

G. LED Power

H. Phase Contrast

I. Brightfield

J. Installing Objectives

K. About Lu aview

L. Downloading and Installing Lu aview

M. Connecting the LS Microscope

N. Live Cell I aging with LS Microscope in an Incubator

II. Software: Getting Started with Lu aview

A. Starting Conditions

B. Launching Lu aview

C. Initialization and Calibration (LS720 only)

D. Instru ent Setup

E. Finding a Live I age With Manual Focus

F. Focusing (Manual on LS720)

G. Snapping I ages

H. Generating Auto Co posite I ages

I. Videos Fro Ti e-Lapse and Live Video

J. Setting Up Autofocus (LS720 only)

K. Writing and Using Protocols

L. Creating a Z-Map

M. Editing a Z-Map

N. Tiling Across Locations

O. Live Video

P. Run Protocol

III. Appendices

A. Red Shipping Lock for LS720

B. Uninstalling a Different Version of Lu aview

C. Disinfecting the LS Microscope Before Place ent into an Incubator

D. Resolving Lu aview Driver Conflicts

E. Lu aview Folder File Organization

F. Capturing and Co piling Live Video Using AviSynth and MeGUI

G. Setting up Windows 10 for Uninterrupted Ti e-Lapse

LS Microscope Startup Manual 2018.7.25

This docu ent is available for download at http://etalu a.co /products/downloads. Any steps specific for a

particular LS odel are noted in italics.

I. Setting Up the LS Microscope

A. Items Included With Each LS Microscope

1. LS720

•USB co unication cable

•External power supply/cord with country-specific plug

•Hex wrench (3 ) for re oving/replacing shipping lock

•Fluorescence shroud

•Calibration 1536 well icroplate

•Calibration 4x objective

2. LS620 and LS600

•Brightfield Illu inator LS620/LS560/LS460

•USB power/co unication cable

•Fluorescence shroud

•External power supply/cord with country-specific plug LS620/LS560

3. LS560 and LS460

•Brightfield Illu inator LS620/LS560/LS460

•USB power/co unication cable

•Fluorescence shroud (LS560 only)

•External power supply/cord with country-specific plug LS620/LS560 (LS560 only)

B. ptional Accessories (purchased separately)

•Phase Contrast Accessory LS720 (Oly pus)

•Phase Contrast Accessory LS620/LS560/LS460 (Oly pus)

•Manual XY Stage LS620/LS560/LS460

•Labware Holders

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

Holder for 60 Petri dishes & Terasaki plates, SBS outer di ensions

Holder for icroscope slides & 50 Petri dishes, SBS outer di ensions

Holder for 4 icroscope slides in parallel, SBS outer di ensions

•Light Hood

•Lu aquant, i age analysis software

•Microvolution, deconvolution software

C. Recommended Computer Specifications

All LS Microscopes: Windows 7, 8, 8.1, or 10; Core i5 or better processor; 500 GB to 1 TB hard drive, 4 GB

ini u RAM; single 4K onitor or two with 1080p HD resolution. Windows ust have Service Pack 2.0

with .NET version 4.5.2.

LS Microscope Startup Manual 2018.7.25

LS720 only: Due to autofocus, Z-stacking, and tiling, a solid-state hard drive (SSD) is required.

Microvolution deconvolution software: A GPU is required; the ost powerful NVidia chip running CUDA

that can be afforded is reco ended for best perfor ance.

D. Placing the LS Microscope

1. Re ove LS Microscope fro the shipping box (save box and packing aterials) and place on a sturdy

bench or counter.

2. If the LS Microscope will be used in an incubator or environ ental cha ber, it is reco ended that

the initial setup be carried out in a roo with a bient conditions before place ent into the

specialized environ ent. See Section I.O. for infor ation about live cell i aging in a specialized

environ ent.

E. Removing the Shipping Lock (LS720 only)

WARNING: You must re ove red shipping lock before plugging the LS720 into

power! If not re oved, the LS720 will be da aged! (See Appendix A for full

details.)

1. Using the supplied 3 hex wrench, loosen the 2 set screws on top of the

installed shipping lock. Loosen the 2 set screws (s aller and ore internal) on

botto of the lock.

2. Unscrew co pletely the 2 larger 1-inch screws on botto of the shipping lock.

Slide the shipping lock toward you to re ove.

3. Keep the 4 set screws screwed into the shipping lock and place in a secure place along with the two 1-

inch screws and 3 hex wrench.

WARNING: If packing the LS720 for ship ent, you must install the red shipping lock before placing into

the shipping carton! If not installed, the LS720 can be da aged! See Appendix A for instructions.

F. Connecting the Cables

1. Wait to connect the USB cable fro the LS Microscope to the co puter until Lu aview is installed

(see Sections I.M-N. below).

2. (LS720 Only). Plug the external power supply/cord into the left side port (when viewing fro the

front) and the plug into an AC outlet.

G. LED Power

1. (LS720 only). The external power supply/cord also supple ents power co ing fro the co puter via

the USB cable to power the LEDs at a higher level than if connected by the USB cable only.

LS720 Shipping lock

LS Microscope Startup Manual 2018.7.25

3. (LS620 and LS560 only). For al ost all fluorescence i aging applications, power fro the co puter

via the USB cable to the LS Microscope will be sufficient. If higher levels of illu ination are required,

e.g., with very di sa ples, insert the s all round plug of the external power supply/cord into the

round port labeled SUPPLEMENTAL P WER on the back of the LS Microscope and the electrical plug

into an AC outlet. You should see a oderate increase in the Illu ination level.

H. Phase Contrast

1. LS720 only

a. If using the Phase Contrast Accessory LS720 (Phase

Accessory), re ove fro its shipping box (save box

and packing aterials). Attach Phase Accessory by

placing the holes on the botto of the Phase

Accessory ar over the pegs on the upper surface

(right rear) of the LS720. Make sure pegs are fully

seated and ar is totally flush with the LS720

surface. Tighten the large thu b screw at the back

of the Phase Accessory ar .

b. Connect the free end of the Phase Accessory co unication cable to the round port on the LS720

back panel (left side) labeled External La p Power Out. This allows the Phase Accessory to be

controlled by Lu aview.

2. (LS620, LS560, LS460 only)

a. If using the Phase Contrast Accessory LS620/560/460 (Phase Accessory),

re ove fro its shipping box (save box and packing aterials). Ensure that

the Phase Accessory adaptor is attached to the rear tab of the LS

Microscope Deck. Attach Phase Accessory by inserting the base of the ar

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

thu b screw on the botto of the adaptor.

b. Connect the free end of the Phase Accessory co unication cable to the

round port on the LS Microscope back panel (left side) labeled External La p

Power Out. This allows the Phase Accessory to be controlled by Lu aview.

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 visible and typically present in the iddle position. Any

open positions can be used for Brightfield.

I. Brightfield

1. For opti al brightfield, use of the Phase Accessory (Section I.H. above) with the included Slider in an

open position (no phase ring) is reco ended. The Phase Accessory, which provides colli ated light,

Phase Accessory cable

Phase Accessory ar

attach ent

Inserting Slider

Phase Accessory adapter

LS Microscope Startup Manual 2018.7.25

is directly connected to the LS Microscope, allowing it to be controlled by Lu aview. It should also be

used when i aging in a dark environ ent such as an incubator. To ount the Phase Accessory on an

LS Microscope, see Sections I.H. above. Brightfield i ages will be in gray scale due to the

onochro atic CMOS ca era.

2. If the Phase Contrast Accessory is not available, a bient light in a typically lit area can often be used.

The light should be as unifor as possible to prevent shadows or other dark or light areas. Overhead

fluorescent lighting can cause uneven brightness including a striped pattern on the i age; if observed,

ove the LS Microscope or partially shade the light to reduce unevenness across the sa ple.

J. Installing bjectives

1. LS720 only

a. Unscrew the deck knob at the front of the instru ent (side with LS720 logo)

and raise the top deck (the top layer of the hinged 2-layer deck). Tipping the

top deck backward will latch it into place. Unscrew the black cap (or

previously installed objective) on top of the optics block and screw in the

objective. Do not overtighten. To lower the top deck, activate the release

lever, lower the deck, and tighten the white deck knob.

2. LS620, LS560, LS460 only

a. Manually re ove the insert of the top accessible 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 re ovable insert to the deck.

3. When installing or changing an objective, carry out the insertion quickly to ini ize dust falling onto

the irrored dichroic filter. A s all a ount of dust will not affect i aging. However if dust is

significant and it appears to affect quality of the live i age or snapped i ages, contact Etalu a by

e ail (support@etalu a.co ) or telephone (760-298-2355) to discuss possible ethods for cleaning.

IMPORTANT: Using co pressed air can result in an oily liquid being sprayed onto the dichroic, which

ay da age it. Si ilarly use of physical ethods such as swabs or lens paper ay also da age the

dichroic and ust NOT be used.

K. About Lumaview

1. LS Microscopes LS720, LS620, LS600, LS560, and LS460 are controlled by the Lu aview software

progra . The latest Lu aview version is downloadable fro Etalu a’s website and must be installed

prior to connecting your computer for the first time to the LS Microscope (due to the need for two

drivers that are installed on the co puter at the sa e ti e).

2. Lu aview will run on Windows 7, 8, 8.1, and 10. Both desktop co puters and laptops can be used,

but the best visualization correlates with onitor resolution equal to the sensor resolution (up to 1900

x 1900 pixels). Note: The onitor does not affect i age resolution unless the onitor is low quality

and affects your ability to judge focus.

The co puter should have Windows .NET Fra ework 4.5.2 or higher installed; see

https:// sdn. icrosoft.co /en-us/library/bb822049%28v=vs.110%29.aspx for ore infor ation. If

Deck release lever

LS Microscope Startup Manual 2018.7.25

your co puter does not have .NET Fra ework 4.5.2 or higher installed, downloading Lu aview ay

auto atically take you to the Microsoft .NET download page. Scroll down the page to find the correct

download. This should not require a purchase. Contact Etalu a with any questions.

L. Downloading and Installing Lumaview

1. To download Lu aview, go to http://etalu a.co /products/downloads (under the Resources tab).

Click on Lu aview - ZIP link to start the download and save the folder when pro pted. Go to your

downloads location and click to open the Lu aview_v(version#).zip file.

Alternatively, the sa e Lu aview_v(version#).zip file can be copied fro the flash drive that co es

with the LS Microscope. Verify that this file is the latest version as posted on the Etalu a website.

2. Lu aview will not run on a co puter with Lu aview 500/400 installed. To uninstall, use the

procedure described in Appendix B.

If your co puter has an older version of Lu aview or Lu aview 720/600-Series (latter no longer

used), installing the newest Lu aview will overwrite the older versions.

If there is a need to revert to an older version Lu aview, it will be necessary to first uninstall the

current version using the procedure described in Appendix B.

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

. si installer file to start the installation. If a Windows warning box about an unrecognized App

appears, click Run anyway.

You will be asked about the location; note that the default is a new Etalu a folder inside the Progra

Files (x86) folder. During installation, a Device Driver Installation Wizard will open. Click to continue

(twice) and finish installing the two drivers. Installation of Lu aview will then finish. After installation,

a Lu aview shortcut (orange circle icon) convenient for launching the software will be present on the

desktop.

M. Connecting the LS Microscope

1. If connecting with a co puter that has been off, turn co puter on. Make sure co puter is connected

to its onitor(s). Insert the standard USB-A end of the supplied USB cable into a USB port on your

co puter and the other square USB-B end into the square port on the LS720 left side or the square

port labeled T C MPUTER on the back of the LS620, LS560 and LS460. The sa e USB port on your

co puter should be used every ti e you connect to the LS Microscope. It is also i portant to connect

the LS Microscope directly to the co puter USB port and not use a USB hub.

2. If Lu aview has not been run previously, it is i portant that the LS Microscope and co puter be

connected using the USB cable before launching Lu aview. This is because Windows needs to load

the USB drivers before Lu aview can run. After the first ti e, the progra can be launched before

connecting the LS Microscope and co puter. In the latter case, only the Outer Main Window will

open (see Section II.B. below). Connecting the LS Microscope and co puter with the USB cable will

result in opening of the Live I age Window as well.

N. Live Cell Imaging with LS Microscope in an Incubator

LS Microscope Startup Manual 2018.7.25

1. Prior to i aging in an incubator, place the LS Microscope inside for at least 6 hr and preferably

overnight to allow ther al equilibration. Before transferring to the incubator, disinfect the LS

Microscope using any of the suggested procedures in Appendix C. Condensation on the icroscope,

including on the objective, will for in the first inutes but will evaporate as ther al equilibriu is

achieved. Focus levels ay change during this equilibration as aterials expand.

2. Pass the co unication and optional power cable through the incubator port, which is often located

in the upper back wall. A plug or filtered stopper ay be present. Every LS Microscope now co es

with a foa plug that can be used to seal around the cable(s) going through the port. Or the existing

plug can be adapted as well. Make sure there is enough slack in the cables inside the incubator for

sliding out the shelf. Place the co puter to be used on a flat and stable surface near to the incubator

and within the 3 M USB cable provided.

WARNING: Do not extend the USB cable or use another USB cable other than that supplied,

particularly a longer one, as co unication issues will arise.

3. LS620, LS560 and LS460: Place the sa ple on the icroscope, adjust focus and close the incubator

door. Check the focus after 60 in to confir it is still good. If the focus has changed, refocus and

repeat. For higher agnification objectives (40x and above), a focus drop of a few icrons ay occur,

which can be co pensated for by over focusing to a height above the focal plane equal to the

expected drop. In this case, the focus will drop to the correct level within a few inutes.

4. LS720 only: Place the sa ple in the labware carrier and ensure it is seated flat with the upper left

corner fir ly pushed into the fra e. Close the incubator and return to Lu aview to focus. The

otorized Z axis will aintain focus level and autofocus can be used to adjust any change in focus over

ti e.

5. The syste is now ready for live cell i aging.

LS Microscope Startup Manual 2018.7.25

II. Software: Getting Started with Lumaview

The following sections give instructions for getting started with Lu aview on the LS720, LS620, LS560 and

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

explained in any particular section. The co plete User Guide for Lu aview is contained in the Help Section

after Lu aview is opened. Click on Help in the Title Bar to open the pulldown enu and click Contents, or

si ply 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. Lu aview has been downloaded and installed on the co puter.

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

3. LS620, LS560, or LS460: Sa ple to be i aged is present on the LS Microscope stage.

4. LS620 or LS560: Fluorescence shroud is over sa ple if i aging involves fluorescence.

5. LS720: Red shipping lock MUST be off, power cable is plugged into a standard AC outlet, and fluorescence

shroud is available if i aging involves fluorescence.

B. Launching Lumaview

1. Launch the Lu aview software fro the desktop icon (or other chosen location) and allow the LS

Microscope to be discovered. This ay take a inute or so the first ti e. Two windows will open on

your onitor:

a. Live I age Window inside the larger Main Window. “Live I age Window” will be in the title bar

when the Window is not axi ized. When Live I age Window is axi ized, [Live I age

Window] transfers to the end of the Main Window title bar.

The Live I age Window status bar at the botto shows co unication with the ca era sensor

via continuous display of several useful etrics: current fra e rate, data transfer rate, nu ber of

fra es collected thus far in the session, fra e size, and session date and ti e start.

b. Main Window with Lu aview v(version nu ber) in the title bar. When Live I age Window is

axi ized, the title bar beco es Lu aview v(version nu ber) – [Live I age Window].

Note: If Live Image Window is not present, there is likely a Driver issue. Follow procedure on

Driver Resolution in ppendix D.

2. The Live I age Window includes a Left Toolbar with the axi u following co ands:

a. Manual I age

b. Manual XYZ (LS720 only)

c. Snap I age

d. Co posite (LS720 and LS620 only)

e. Video Record (toggles on and off)

f. Protocol

g. Zoo

LS Microscope Startup Manual 2018.7.25

h. Last File

When Lu aview is open, the icon that resides in your co puter dock appears as a docu ent with an

arrow.

C. Initialization and Calibration (LS720 only)

1. The LS720 auto atically initializes every ti e Lu aview is started by oving the icroplate carrier to

the Ho e position. The Ho e Position consists of the left front corner of the icroplate carrier fra e

being positioned over the objective and the objective retracted to the lowest Z level. No i age can be

obtained at the Ho e position because it is under the icroplate carrier fra e.

2. If this is the first ti e the LS720 has been connected to the current co puter, it is first necessary to do

a Calibration. Once co pleted, calibration is not needed unless a different co puter

is connected or the XY position appears to be different fro the expected position,

e.g., center of icroplate wells.

a. To calibrate the LS720, install the provided Calibration Motic EFN 4x Objective.

Place the provided Calibration 1536 well icroplate into the icroplate carrier.

b. Click Utilities in the File Menu bar and in the drop-down enu click Calibrate to

open its dialog box. A co unication dialog box will open asking if a 4x objective

has been installed. Click Yes and the co unication dialog box will close.

c. The Live I age Window will now show red crosshairs, and the icroplate carrier

will ove so well A1 is close to being over the objective.

d. Click the Manual I age icon in the left tool bar (top icon) to open its dialog box.

e. To use a bient light for calibration (reco ended; see Section I.I.), check

Brightfield in Manual I age to activate Gain and Exposure sliders. Start with

Gain as low as possible and adjust Exposure until a id-gray tone is shown in

the Live I age Window.

f. If using the Phase Contrast Accessory for brightfield illu ination (Section I.I.),

ove the phase slider to an open position (without phase ring) and ove the phase condenser

aperture (s all lever on condenser front above slider) so only a low level of light

is trans itted. In Manual I age, check Brightfield, set Gain to ini u ,

Illu ination (%) to ini u , and adjust Exposure until a id-gray tone is seen in

Live I age Window.

g. In Calibration, ove the Focus slider up while watching the Live I age Window

to detect when the well outline can be seen and is in sharp focus. The Focus

slider area beco es green whenever the Focus slider is active. Using the X/Y

Move arrows (and Step size arrows if needed), ove the icroplate carrier until

the A1 well is centered in the Live I age Window. Make sure the centered well

Red crosshairs in Live

I age Window

Otros manuales para LS720

Este manual sirve para los siguientes modelos

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