AMG EVOS fl User manual
© 2010 Advanced Microscopy Group. All rights reserved. Doc Control Number ZP-PKGA-0495
REV B Published 20 SEP 2010
EVOSfl USER GUIDE
SETUP ............................................................................. 3
Standard Items Included............................................... 3
Moving/Transporting EVOS.......................................... 4
Operating Environment.................................................4
Mechanical Stage...........................................................4
LED Light Cubes ............................................................ 5
Power Supply ................................................................. 5
USB Ports........................................................................ 5
DVI Output Port ............................................................. 5
UV Shield ........................................................................ 6
Arm Rest (optional)................................................ 6
Installing EVOS in a Cell Culture Hood........................ 7
QUICK-REFERENCE DIAGRAMS .................................. 8
BASIC OPERATION........................................................9
Fluorescence Operation................................................ 9
Brightfield or Phase Contrast Operation ....................11
ADVANCED OPERATION .............................................13
Logging In/Creating New User Logins.......................13
Saving Images & Working with Files ..........................14
Using the QuickSave Option .......................................15
Recording Time Lapse Images....................................16
Using the Transfection Tool .........................................17
Counting Cells...............................................................18
Reviewing Images.........................................................19
Using Digital Zoom......................................................20
Connecting EVOS to a Network ..................................21
Changing LED Light Cubes......................................... 23
Updating Software ......................................................24
CONTROLS GLOSSARY ...............................................25
Onscreen Controls ....................................................... 25
Mechanical Controls ....................................................29
CARE & MAINTENANCE...............................................31
General Care..................................................................31
Objective Lens Care......................................................31
Stage Care .....................................................................31
Sterilization Procedures...............................................31
TROUBLESHOOTING...................................................32
Image Quality Issues ................................................... 32
Software Interface Issues............................................ 32
Mechanical Issues ........................................................ 33
CUSTOMER & TECHNICAL SERVICE..........................34
AMG Contact Information...........................................34
EVOS Distributor Information ....................................34
Service and Warranty Information ...........................34
PARTS & ACCESSORIES ..............................................35
SPECIFICATIONS .........................................................36
WARNINGS, PRECAUTIONS & NOTICES
Throughout this manual, the following types of notifications require your close attention:
!WARNING! This type of notification tells how to avoid serious personal injury.
IMPORTANT! This type of notification tells how to avoid damaging the microscope and/or voiding your warranty.
(Contact your local distributor for more warranty information.)
This symbol indicates information specific to the color camera version.
MONOCHROME CAMERA VS. COLOR CAMERA
The EVOS fl microscope is factory-configured with either a monochrome camera or a color camera. Monochrome
cameras are commonly used for high-performance fluorescence applications, and provide the best sensitivity for
detection of faint fluorescence signals. Color cameras have lower fluorescence sensitivity but have the advantage of
being able to dierentiate structures by color in transmitted light, e.g., imaging stained tissue samples. Throughout
this User Guide, any operational dierences between the two versions of the microscope have been noted.
CONTENTS
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STANDARD ITEMS INCLUDED
Before setting up your new EVOS, unpack the unit and
accessories and verify all parts are present. Contact your
distributor if anything is missing.
Note: If you do not have your distributor information, you
can look it up at the AMG website or contact AMG
Customer Service (see p. 34).
EVOSflmicroscope, per order
Condenser shield, removable
Power adaptor
Dust cover
Mouse pad
Quick Start Guide
USB flash drive (includes User Guide)
Accessories boxes
Smaller box
đƫ Light shield box
đƫ USB mouse
đƫ Power cord
Larger box (for storage)
đƫ UV shield assembly (mount, shield, screws &
L-shaped hex key)
đƫ Light cube access cover (includes LED light cube
installation tool); install light cube access cover
before using EVOS
đƫ Condenser sliders: Pinhole, Diuser, Meniscus A,
MeniscusB, and Block
LED light cube lock
(in place under stage)
LED light cubes (in place under
LED light cube lock)
Light shield boxUV shield assembly kit
Light cube access cover (top) Light cube access cover
(bottom, with tool)
Condenser shield, removable
SETUP
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SETUP
MOVING/TRANSPORTING EVOS
ALWAYS lock stage with the stage lock pin before
moving microscope.
When transporting or shipping EVOSfl , secure the LED
light cubes in place with the light cube lock .
Lift the microscope by grasping it firmly with both
hands under the support arm , balancing the weight
as shown at left.
To transport EVOS to a dierent facility, use the original
packaging materials if possible. Always be sure the
microscope is properly cushioned and braced to prevent
damage.
IMPORTANT! Never allow EVOS to be subjected to
sudden impact or excessive vibration. Handle the
microscope with care to prevent damage.
OPERATING ENVIRONMENT
Place the microscope on a level surface away from
vibrations from other pieces of equipment.
Allow at least 5 cm (2 in) free space at the back of the
microscope to allow for proper ventilation and prevent
overheating of electronic components.
Set up EVOS away from direct light sources, such as
windows. Ambient room lighting can enter the imaging
path and aect the image.
Note: Place the light shield box on the stage over
the sample to reduce the eects of ambient
light and improve image quality.
Operating temperature range: 4°–32°C (40°–90°F).
Relative humidity range: 30–90%.
IMPORTANT! Never subject EVOS to UV sterilization.
UV degrades many materials, including plastic.
Damage from UV exposure is not covered under the
manufacturer’s warranty.
MECHANICAL STAGE
STAGE LOCK PIN
Before moving the mechanical stage for the first time,
remove the stage lock pin from the back right-hand
corner of the stage plate. Pull firmly to remove this pin.
You may store the stage lock pin in your accessories box
for future use.
Note: Always secure the mechanical stage with the stage
lock pin before moving the microscope.
Grasp under support arm
with both hands to lift EVOS
3
Light shield box on stage
4
Stage lock pin engaged
1
Light cube lock engaged
2
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SETUP
LED LIGHT CUBES
IMPORTANT! Before changing light channels, ALWAYS
be sure the light cube lock has been removed. Applying
force to the light cube selection lever while the lock is
in place may seriously damage the mechanism. This
type of damage is not covered by the manufacturer’s
warranty.
1. Move the stage back to allow access to the light cube
lock , which is centered under the back of the stage.
2. Loosen the thumbscrew to remove the light cube lock.
You may store the light cube lock in your accessories
box for future use during transport or shipping.
3. Place the light cube access cover into the opening
and tighten thumbscrew.
Note: For information about adding optional LED light
cubes, refer to Changing LED Light Cubes (p.23).
POWER SUPPLY
1. Turn the power switch to the “” (OFF) position
before connecting the power adaptor.
IMPORTANT! Always use the correct power supply.
The power adaptor specifications appear on the
serial number label (front of LCD hinge) and in the
SPECIFICATIONS (p.36).Damageduetoanincompatible
power adaptor is not covered by warranty.
2. Connect the power adaptor to the power jack on the
right side of the microscope base, attach the cord to the
adaptor, and plug the cord into an outlet.
USB PORTS
Plug the mouse and the USB flash drive into the USB ports
located on the bottom right of the support arm. You may
also plug in a USB keyboard (not included) for text input.
DVI OUTPUT PORT
A DVI port is available for output to a projector or other
display. (DVI cable not included.) This port produces
digital output only; EVOS is compatible with either a DVI-D
or DVI-I display.
Power switch Power adaptor plugged in
LED light cube lock engaged
1
Remove light cube lock Secure light cube access cover
over opening
12
Mouse and USB flash drive
plugged in
DVI cable (not included)
plugged in
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SETUP
UV SHIELD
!WARNING! UV LIGHT HAZARD! This microscope uses
a Class 3B ultraviolet LED for the DAPI channel. Avoid
exposure to the UV beam and use protective shields.
NEVER look directly at light.
For your protection, follow this procedure to install the UV
safety shield before using the DAPI fluorescence channel.
1. Attach the UV shield mount to the front of the
condenser with the two screws provided.
2. Remove the protective coverings from both surfaces of
the UV shield .
3. Place the holes in the UV shield over the screws on the
mount and push the slots down on the screws to secure
the shield in place, as shown.
Note: The UV shield is removable for access to the
condenser sliders used in transmitted light
mode. Simply unhook it from the screws on the
UV mount.
ARM REST (OPTIONAL)
An optional arm rest kit is available. (See PARTS &
ACCESSORIES (p.35) for ordering information.) The arm rest
fits on either side of the stage. To attach the arm rest, it
is necessary to remove a Y-axis stage knob (right knob
shown). Store this knob in your accessories box or other
safe place.
1. Use the smaller L-shaped hex key supplied in the arm
rest kit to loosen the 2 stage knob screws .
Note: The Y-axis stage brake is on the left knob. If you are
placing the arm rest on the left side, remove the right
knob and replace it with the knob that has the stage
brake. The stage brake is useful for time lapse captures.
2. Slide the stage all the way to the side opposite the
intended arm rest position. This will expose the holes
for the arm rest screws.
3. Align the arm rest base slots over the holes as desired
and use the larger hex key to attach the base to the stage
with the arm rest screws .
4. Adjust arm rest height, if desired:
a. Pull the adjustment pin almost all the way out to
allow the arm rest post to move up or down, and
set the arm rest to the preferred height.
b. Push the adjustment pin in. It may be necessary to
move the arm rest slightly so the pin can fit through
the groove on the post.
UV shield in place
3
Attach UV shield mount
1
2
Stage brake on left Y-axis knob
3
Slide stage to expose holes
4
Attach arm rest to microscope
5
Remove Y-axis stage knob
1
2
Adjustment pin and arm rest
post
7
6
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SETUP
INSTALLING EVOS IN A CELL CULTURE HOOD
EVOS’ small footprint, simple power connection, and
easily-viewed display make it quick to install and
convenient to use in a cell culture hood.
DIMENSIONS
EVOS will fit in cell culture hoods that are at least 20 ½
inches (520 mm) deep. If your cell culture hood is smaller,
it may be possible to turn the EVOS at a slight angle to fit.
EVOS installed in a cell culture hood
LCD tilted back into transport position
INSTALLATION
Note: Refer also to the illustrations on p. 4 for more
details about moving EVOS.
1. Secure the stage with the stage lock pin, switch EVOS o,
and disconnect the power cord, mouse and, if connected,
keyboard.
2. Tilt the LCD screen back until it is parallel with the
tabletop.
3. Lift the microscope by grasping it firmly with both
hands under the support arm just behind the condenser.
4. Gently place the microscope on a lab cart and transport
it to the cell culture hood.
Note: Verify that the hood sash is raised enough
for the microscope to slide underneath
(approximately 14” or higher).
5. Lift the microscope as before and move it into the hood.
6. Tilt the LCD monitor upright.
7. Remove the stage lock pin, connect the power cord,
mouse and, if desired, keyboard, and switch EVOS on.
REMOVAL
Reverse the sequence above to remove EVOS from the cell
culture hood.
ENGLISH METRIC
DEPTH 18.5 in 47.0 cm
WIDTH 14.0 in 35.5 cm
HEIGHT, TRANSPORT12.75 in 32.4 cm
HEIGHT, DISPLAY22.75 in 57.8 cm
WEIGHT 33.7 lbs 15.3 kg
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QUICK-REFERENCE DIAGRAMS
Note: Refer to the CONTROLS GLOSSARY (p.25) for more details about onscreen and mechanical controls.
1. Channel indicator bar
2. Active channel
(highlighted)
3. Login button
4. Control bar
5. Control bar tabs:
đƫ Find & Focus
đƫ Actual
đƫ Overlay
6. LIGHT ON/OFF button
7. Illumination slider
8. Exposure time slider
9. Image capture button
10. Color/boost options
11. Scalebar/toolbar
options
12. Settings control button
13. Save image button
14. Info display bar*
15. Selected objective
1
8
54 6 9 10 1311 14127 15
2
67
9
1. Power switch
2. Power input jack
3. USB and DVI ports
4. Coarse stage positioning
knobs
5. Stage X-axis knob
6. Stage Y-axis knobs
7. Focusing knobs
8. Objective selection wheel
9. Light cube selection lever
10. Phase annuli selector
11. Condenser slider slot
3
6
7
8
5
71
4
10
10
11
11
3
*The color camera version shows the QuickSave option instead of the info display bar.
1
7
2
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The EVOSflmicroscope uses both mechanical and software
controls for operation. Mechanical controls include the
stage X-Y axis knobs, focusing knobs, objective selection
wheel, and the light cube selection lever. Software controls
are located in the control bar at the bottom of the
screen. The channel indicator bar at the top of the
screen shows the selected filter cube or transmitted light
position. The login button displays the current user ID.
Refer to the QUICK-REFERENCE DIAGRAMS (p.8) as needed.
See also the CONTROLS GLOSSARY (p.25) for more details.
FLUORESCENCEOPERATION
1. Turn on the microscope using the power switch on
the right side of the microscope base.
2. Plug a USB flash drive into one of the USB ports on
the right side of the microscope arm.
3. Place the sample on the stage, using a vessel holder if
needed.
Note: Place slides with coverslips face up.
4. Set the magnification using the objective selection
wheel on the front of the microscope.
5. Move the light cube selection lever on the left side
of the microscope all the way toward you. (The channel
bar will highlight the “Transmitted” position.)
6. Turn on illumination using the LIGHT ON button ,
located on the left side of the control bar.
7. Focus the sample using the focusing knobs .
8. Place the light shield box 10 on the stage, over the
sample. This is important for optimal image quality.
Note: If your application requires access to the sample,
work in a dark room and use the Block slider to
block light reflected from the condenser.
9. Move the light cube selection lever to the desired
fluorescence channel. (The channel indicator bar will
highlight the selected light cube.)
10. Click the Find & Focus tab.
11. Click the LIGHT ON button to turn on fluorescence
illumination.
! WARNING! UV LIGHT HAZARD! When using the DAPI
channel, avoid exposure to beam and use protective
shields. NEVER look directly at light.
12. Adjust the focus as needed.
13. Adjust the illumination intensity if necessary, using the
Illumination slider on the control bar or the mouse
scroll wheel.
Light cube selection lever
10
Light shield box 10 in place on stage
Power switch and data ports
LIGHT ON button in the control bar
8
1
2
Software control bar , channel bar and login button
3
continued on next page
BASIC OPERATION
7
7
4
5
4
Objective selection wheel and focusing knobs
6
9 9
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BASICOPERATION
Note: When the Color option is o, overexposed
pixels will appear red. Dim the illumination
until the red highlights disappear to get the
maximum level of brightness without any
overexposed areas. See p. 27 for instructions on
changing the overexposed pixel display.
For the color camera version, the overexposed
pixels are always highlighted in red unless this
feature is disabled in the settings.
14. Click the Capture button to acquire the image.
15. Move the light cube selection lever to the next position
and repeat steps 10-14 to acquire each fluorescence
channel as desired.
16. Click the Overlay tab to show all channels in color
overlay.
17. Adjust brightness and contrast for each channel to bring
them into balance with each other.
18. Click the Save button to save the color image. The
Save File dialog box will pop up.
19. Click in the Save File Name text field to enter the file name.
A virtual keyboard will pop up. After entering the file name,
click the Accept button at the lower right of the keyboard.
20. Choose the file type and click the Save button .
Note: See Saving Images & Working with Files (p.14)
for more information.
HELPFUL TIPS
The Boost option 10 amplifies faint signals near the
background. This only changes the display; it does not
alter the data.
Turn othe Color option to display the image in grayscale.
This often shows more detail than a color image.
Find & Focus uses a shorter exposure time (100 ms)
and lower illumination (appx. 60%) compared to image
capture settings. This minimizes photobleaching and
phototoxicity eects. When you capture an image,
illumination and exposure time automatically adjust for
best image quality and then reset to lower levels after
the capture.
The Actual tab provides full-powered illumination and actual
exposure times for live viewing of the sample.
Note: With longer exposure times (more than 200ms)
there will be a lag between focusing the image
and seeing the focus change onscreen.
Actual tab
Illumination slider and Capture button in the control bar
1 3
Color and Boost 10 options; Save button
10 5
2
Overlay tab
4
Virtual keyboard
7
Save File dialog box
6
8 9
Fluorescence Operation, continued
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BASICOPERATION
BRIGHTFIELD OR PHASE CONTRAST
OPERATION
1. Turn on the microscope using the power switch on
the right side of the microscope base.
2. Plug a USB flash drive into one of the USB ports on
the right side of the microscope arm.
3. Place the sample on the stage, using a vessel holder if
needed.
Note: Place slides with coverslips face up.
4. Set the magnification using the objective selection
wheel on the front of the microscope.
5. Move the light cube selection lever on the left side
of the microscope all the way toward you. (The channel
indicator bar will highlight the “Transmitted” position.)
6. Turn the phase annuli selector to the position that
corresponds to the selected objective and contrast
method.
7. Insert the appropriate condenser slider into the slot
on the condenser assembly for optimal image quality.
Five condenser sliders are included with EVOSfl
microscopes. For brightfield applications you may use
the Diuser or Pinhole slider, or a Meniscus slider.
Due to variances in sample size, color and thickness,
actual slider use may dier from the suggested slider
use described below.
Diuser slider: Brightfield, 2x or 4x (for flat field
illumination)
Pinhole slider: Brightfield, all magnifications (to
enhance contrast)
Meniscus A slider: Brightfield, 2x (for low-volume fluid
in a multi-well dish)
Meniscus B slider: Brightfield, 2x (for high-volume fluid
in a multi-well dish)
8. Turn on illumination using the LIGHT ON button ,
located on the left side of the control bar.
9. Focus the sample using the focusing knobs .
Phase annuli selector
continued on next page
Insert slider into condenser
6
8
Objective selection wheel and focusing knobs
8
3
5
BF
Light cube selection lever
4
2
Power switch and data ports
1
4
LIGHT ON button in the control bar
7
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BASICOPERATION
10. When switching from fluorescence to transmitted light
with the light shield boxon the stage, remove the
light shield box cover so the light from the condenser
can pass through.
11. Adjust the illumination intensity if necessary, using the
Illumination slider on the control bar or the mouse
scroll wheel.
Note: Overexposed pixels will appear red. Dim the
illumination until the red highlights disappear
to get the maximum level of brightness without
any overexposed areas. See p. 27 for instructions
on changing the overexposed pixel display.
12. Click the Capture button to acquire the image.
13. Click the Save button to save the image. The Save
File dialog box will pop up.
14. Click in the Save File Name text field to enter the file
name. A virtual keyboard will pop up. After entering the file
name, click the Accept buttonat the lower right of the
keyboard.
15. Choose the file type and click the Save button .
Note: See Saving Images & Working with Files (p.14)
for more information.
1
Light shield box in place on stage (cover shown in place)
Illumination slider and Capture button in the control bar
4
Save button
5
Virtual keyboard
7
Save File dialog box
6
98
Brightfield or Phase Contrast Operation, continued
3
2
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Virtual keyboard
LOGGING IN/CREATING NEW USER LOGINS
EVOS keeps settings in memory for each user ID, so multiple
users can work with the same EVOS microscope without
having to reset their preferences.
To use this feature, set up a user profile for each regular user.
You may also assign user IDs for experiments in progress.
Note: User profiles are not password protected. All users
should verify they are logged in correctly to avoid
changing others’ settings.
LOG IN WITH AN EXISTING PROFILE
1. Click the login button at the bottom left of the
screen. (This is the AMG logo with the current user
profile indicated above.)
2. Select the desired user profile and click OK .
Note: No password is necessary to log in.
ADD OR REMOVE A USERPROFILE
1. Click the login button .
2. To copy an existing profile, highlight the profile in the
user list , select the “Copy from ‘name’$” option , and
click the Add button . The virtual keyboard will pop up
so you can name the new profile.
3. To create a new user profile without copying any
settings, deselect the “Copy from” option , click the
Add button , and enter a user name.
4. After adding a new user profile, click OK to log in
under that name and adjust settings as desired. When
you switch o, EVOS will save your settings to memory.
5. To remove a user profile, highlight it and click the
Remove button . A confirmation dialog box will
pop up. Deleting the user profile will remove all its
associated settings from memory.
6. To rename a user profile, click the Rename button
and enter the new name.
CHANGE THE DEFAULT LOGIN
The default user login is Guest; to set the default login
as the last active user, go to the Basic tab of the Settings
dialog box and uncheck the “Default to Guest on
startup”option.
Note: For multiple users, we recommend leaving the “Default
to Guest on startup” option checked.
Login button (set to Guest profile)
1
Login dialog box
5
3
4
6
7
2
Settings: Basic Tab (to change default login)
8
ADVANCED OPERATION
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ADVANCED OPERATION
SAVING IMAGES& WORKING WITH FILES
When you click the Save button , the Save File dialog
box appears. If there is no USB flash drive or network
connection in place, an information message will
appear. Click the Cancel button to clear this message.
In the Save Folder list and the saved files list , selected
items will appear orange. If a USB keyboard is installed,
the virtual keyboard will not appear. Note that pressing the
Enter key on a physical keyboard is like pressing the Save
button in the Save file dialog box.
1. Click in the Save File Name text field . Enter a file
name and click the Accept button .
To overwrite a file, simply select the name of the file
from the saved files list instead of clicking on the
Save File Name text field. A Save As confirmation
dialog box will pop up. It is not possible to recover an
overwritten file.
2. Click on the name of a folder in the Save Folder list to
select the destination for the new image.
3. To create a new folder, first click the name of the parent
folder, and then click the New Folder buttonto enter
a folder name and, if desired, date.
Note: Clicking the Date button anywhere within a
text field will automatically insert the current
date (MM-DD-YYYY) wherever the cursor is in
that field.
4. Select a file format (.tif, .png, .jpg or .bmp) from the File
Type drop-down menu 10 .
Note: To save a 16-bit image, select .tif or .png and
ensure the Scalebar 11 and Color/Boost 12
options are o. File types .jpg and .bmp (as well
as images of all types with the Scalebar, Color, or
Boost options engaged) only save at 8-bit depth.
5. Click in the Comment text field 13 to enter a comment
and date (optional).
6. Click the Save button 14 to save the file.
7. To delete a file or folder, highlight the item on the list and
click the Delete button 15 . A confirmation dialog box will
pop up. It is not possible to recover a deleted file.
8. To rename a file or folder, highlight the item on the list
and click the Rename button 16 . The virtual keyboard
will pop up; you can use the Clear button 17 to reset to a
blank field.
Confirmation popup
Save button , Scalebar option 11 , and Display options 12
1112 1
Virtual keyboard
9
7
17
5
6
8
14
15
16
Save File dialog box
4
13
10
2
Save File information message
3
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ADVANCED OPERATION
USING THEQUICKSAVE OPTION
QuickSave allows you to save multiple images under a
single base file name. Simply specify the settings and select
the QuickSave option (in the Overlay tab), and EVOS will
save each image with a single click of the Save button.
1. Click the Settings button to open the Settings
dialog box, and then select the QuickSave tab.
2. Click in the Base Filename text field and enter a name
that describes the imaging session. The orange “Next”
file name will reflect the information entered.
3. Click in the Count text field to enter the starting
number, if you do not want to start at 1. The orange
“Next” file name will reflect the information entered.
4. Select a file format (.tif or .png) from the File Type
drop-down menu .
5. Click the Browse button to select a destination folder
for the QuickSave files. In the Browse popup, highlight
the desired folder and click OK .
6. To create a new folder, first click the name of the desired
parent folder, and then click the New Folder button .
Enter a folder name and date, if desired. After creating
the new folder, click OK to close the Browse popup.
Note: Clicking the Date button anywhere within a
text field will automatically insert the current
date (MM-DD-YYYY) wherever the cursor is in
that field.
7. Select “Also save each channel separately” 10 to save
multiple channels for each image. This will create up to
five files per captured image, named according to the
following conventions:
BaseName_RGB_0001.tif (Overlay image)
BaseName_channel_0001.tif (where “channel” is the
selected channel, such as GFP, RFP, DAPI, or TRANS)
When optional LED light cubes are installed, the files
will automatically save channels with their names. See
Changing LED Light Cubes (p.23) for more information.
8. Click OK 11 to accept QuickSave settings.
9. Select the Overlay tab and click the radio button for the
QuickSave option 12 .
Note: The color camera version displays the QuickSave
option in all tabs.
10. After acquiring an image with the Capture button, click
Save 13 . The image will be saved as specified in the
QuickSave settings.
Virtual keyboard
9
Settings: QuickSave Tab
2
34
5
6
10
11
QuickSave option radio button 12
12
Settings button and Save button 13
1 13
Browse popup (to select a destination folder)
7
8
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ADVANCED OPERATION
RECORDING TIME LAPSEIMAGES
With EVOS, you can set up your cells and program the
microscope to record time lapse images. To use this feature,
open the Time Lapse tool in the toolbar, specify the settings,
and click Start. You may pause or cancel sessions in progress.
START A TIME LAPSE SESSION
1. Once the specimen is focused and ready, tighten the
stage brakesto prevent the stage from drifting
during the session.
2. Open the Toolbar and expand the Time Lapse tool.
3. Click the Interval text field and enter a value.
4. Choose a unit of measure for the capture interval from
the interval Unit drop-down menu .
5. Click the Duration text field and enter a value.
6. Choose a unit of measure for the duration from the
duration Unit drop-down menu .
Note: The Images fieldshows the total number of
images for the session.
7. To save the session in a folder other than the default
location, click the Browse button to select the
destination.
8. In the Browse popup (shown on p. 15), highlight the
desired folder and click OK.
9. Under File, click the Base Filename text field to enter
a name, and then choose a file type (.png or .tif) from
the File Type drop-down menu 10 .
10. To create a video (.avi) file, select Write Video File 11 .
11. Click the Start button 12 to begin the time lapse session.
EVOS will display the Time Lapse Progress popup as
long as the session is active.
Note: The Review slider 13 lets you review the images
already captured during the current session. The
Play button 14 shows all the images in sequence.
PAUSE AND RESTART A TIME LAPSE SESSION
In the progress popup, click the Pause button 15 to
suspend the time lapse captures. The progress popup will
dim, and a Resume button 16 will replace the Pause button.
Alternatively, to pause and adjust the settings, click the
Find & Focus button 17 . Click the Start button 12 to resume
the time lapse capture sequence, or uncheck the Resume
radio button 18 and start a new time lapse session.
ABORT A TIME LAPSE SESSION
In the progress popup, click the Abort button 19 . A dialog
box will pop up, giving you the option to delete or keep the
files already saved. Clicking Cancel will resume the session.
Toolbar option in
the control bar
2
Toolbar
Time Lapse Progress paused
16 19
Time Lapse tool
3
5
6
4
7
9
10
12
8
11
Time Lapse Progress popup
15
13
19
14
17
18
X-axis and Y-axis stage brakes
1
www.amgmicro.com 17 EVOSflUser Guide
REV B Published 20 SEP 2010
ADVANCED OPERATION
USING THE TRANSFECTION TOOL
EVOS’ Transfection tool expedites the capture and overlay
of images for transfection analysis.
1. Choose a light cube, focus on the sample, and adjust
the lighting. See steps 1-13 of Fluorescence Operation
(p.9) for detailed instructions.
2. Open the Toolbar and expand the Transfection tool .
Note: The “Pause after first image” option allows
you to adjust focus, if necessary, before
capturing the transmitted light channel.
3. Click the Run Sequence button . The sequence always
starts with the fluorescence channel and finishes with
the transmitted light channel.
Note: The Lighting Override feature allows you to
select a channel to activate with the LIGHT ON/
OFF button.
4. If the Pause option is selected, adjust focus and click the
Continue button . If Pause is deselected, go to step 5.
5. The dual-channel image will display with the Overlay
tab selected. Adjust the brightness and contrast
settings as desired, and save the image . See
Saving Images & Working with Files (p.14) for detailed
instructions.
Sequence paused between images
6
Sample ready for transfection analysis
2
3
4
5
1
Transfection overlay image
7
8
www.amgmicro.com 18 EVOSflUser Guide
REV B Published 20 SEP 2010
ADVANCED OPERATION
COUNTING CELLS
The Count tool streamlines cell counting by marking items with
up to 6 labels onscreen. As you tag items, EVOS will keep a
running tally of counts with percentages for each label assigned.
Document your results simply by saving the tagged image, with
the Count tool displaying the totals.
1. Acquire an image. See BASIC OPERATION (p.9) for
detailed instructions.
2. Open the Toolbar and expand the Count tool .
Note: Use the Add mode , which is active by default.
The Delete mode is for removing tags that
have already been added.
3. Click in a black Label text field to name a label. You
may use up to 6 labels.
4. Under Settings, you may choose a grid size in the drop-
down menu or leave the Show Grid option inactive.
5. Select a Label button and left-click at each point
onscreen to tag the items for that category. Switch
labels as desired; EVOS will tag for the selected label.
Note: To use Digital Zoom while counting cells, first
suspend the Count tool. Either select the Hide
Tags setting , click the triangle to minimize
the Count tool, or click the Toolbar option to
minimize the whole Toolbar. When the Count
tool is suspended, the left mouse button will
behave according to the rules described in
Using Digital Zoom (p.20). After zooming,
reactivate the Count tool by deselecting Hide
Tags or reopening the tool.
6. To move a tag, select 10 and drag it. Left-click anywhere
else onscreen to deselect.
7. To delete a tag, right-click it, or choose the Delete mode
and left-click it. You may also use the Clear All button 11 to
delete all tags for all labels.
8. To save an image showing the labels, counts and
percentages as shown in the Count tool, select the
Save Image with Toolbar option 12 and click the Save
button 13 . Deselecting this option will produce an
image saved with the tags only.
Detail of grid size menu and
Show Grid option
6
7
Selected tag 10 ; drag to move
10
Count tool
3
5
8
9
2
11
12
7
6
4
Toolbar option and Count tool
1
2
13
www.amgmicro.com 19 EVOSflUser Guide
REV B Published 20 SEP 2010
ADVANCED OPERATION
REVIEWING IMAGES
The Image Review tool allows you to review still images or play
video files from the USB drive or network connection. You may
also use this tool to rename or delete saved files.
1. Open the Toolbar and expand the Image Review tool .
2. The preview list displays thumbnail images for all
viewable files in the selected directory. (The top-level
USB directory is selected by default.) If there are no
viewable files in the directory, the preview area will be
empty.
Note: The File Type drop-down menu filters files by
type. By default, it is set to display all files with
.png, .tif, .jpg, .bmp, or .avi extensions.
3. If the image or video file you wish to review is not in the
directory displayed, click the Browse button to find
and open the desired directory.
4. Use the scroll bar as needed to search the preview
list for the desired file. Click the image to select it. The
selected file name appears orange .
5. Click the View File button to display the image in
the image review port . This button toggles between
View File and Hide File. Hiding the file closes the image
review port.
Note: Double-clicking the thumbnail image will also
toggle between displaying and hiding the file.
6. To zoom the image in the review port, double-click
the area of interest; right-click to restore normal
magnification. Refer to Using Digital Zoom (p.20) for
more detailed instructions.
7. To rename a file, select it and click the Rename button 10 .
The Virtual Keyboard will pop up. Enter the new file
name and click the Accept button 11 .
8. To delete a file, select it and click the Delete button 12 .
A confirmation dialog box will pop up. It is not possible
to recover a deleted file.
Image Review Tool: Find and select a file
1
2
36
4
5
7
8
Confirmation popup
Image Review Tool: View, rename or delete a file
8
9
10 12
Virtual keyboard
11
www.amgmicro.com 20 EVOSflUser Guide
REV B Published 20 SEP 2010
ADVANCED OPERATION
USING DIGITAL ZOOM
EVOS can zoom the image onscreen, quickly allowing a closer
look. Simply double-click live or captured images to zoom
them. In the images below, the numbered arrows indicate click
points. Also note that the zoom factor display appears over the
selected objective display.
ZOOM AND RECENTER LIVE IMAGES
Note: Live images can only zoom to 2x. To zoom an image
at higher levels, you must first capture it.
1. Double-click the area of interest in the image
onscreen. EVOS will display a view zoomed 2x,
centered on the point clicked.
2. In the enlarged image, double-click on any point in
the middle area of the screen to recenter the image .
(EVOS will place points from the outer edges of the
screen as close to the center as possible.) You may
recenter repeatedly.
3. Right-click anywhere on the image to restore the view
to unzoomed magnification.
ZOOM CAPTURED IMAGES
1. Double-click the area of interest in the image
onscreen. A view zoomed 2x, centered on the point
clicked, will appear.
2. In the enlarged image, double-click again on any point
to double the digital zoom level. The enlarged image will
center around the point clicked.
3. Continue double-clicking to double the digital zoom
value as desired. It is possible to zoom in to the pixel
level of the digital image.
4. Right-click to restore the view to unzoomed
magnification.
Note: Capturing and saving a zoomed image will result
in a file showing the actual magnification, not the
zoomed magnification. If the scalebar is active, it will
appear in the file.
Captured 40x image zoomed 2xCaptured image at 40x
magnification, with scalebar
3
Image recentered
Live 40x image zoomed 2xLive image at 40x magnification
Captured 40x image zoomed 4x Captured 40x image zoomed 8x
1
2
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