Basic Zeiss LSM 510 Meta User Guide ZEN Software

Basic Zeiss LSM 510 Meta User Guide ZEN Software
Basic Zeiss LSM 510 Meta User Guide
ZEN Software
1. Start-up – Follow the start-up instructions.
1 – Mercury Lamp
2 – Main Switch (left of microscope)
3 – Computer
2. ZEN 2009 icon – double click on this icon to start the software.
3. Start System
4. The Ocular Tab will be selected as it opens
 Pressing the Online button will allow you to view your sample through the oculars
 While in the Offline position the image can be viewed on the computer screen
5. In the Ocular drop menu you will find the microscope positions.
1. Halogen Bulb
2. Aperture
3. Specimen/Stage
4. Objective
o You may select your desired objective here.
5. Reflector Cubes
o Once you’ve selected Online, you may select the filter
that corresponds to the color you’d like to view
through the oculars.
 Rhodamine
 Bright Field Settings
6. Hg Bulb shutter
7. Hg Bulb
8. Lens
6. Select the Acquisition Tab
Under the Setup Manager select the Laser drop
menu as well as the Laser Properties drop menu.
o Argon/2-Select Standby from the drop menu
to warm up the laser. The Status will read
“Ready” once it’s done and you can then
choose On from the drop menu. Adjust the
Output [%] slider so that the Tube Current
reads 6.1 A.
o HeNe1 & HeNe2-Select On
Note: The only lasers necessary to turn on are the
ones you are using. You may leave any unnecessary
lasers in the Off position.
7. Load Specimen – The10x objective uses NO oil or water. There is a 40x oil, 63x oil, and 100x oil objective which all
require a single drop of oil on the lens before loading a specimen.
Place your slide upside down into the stage – the width of the holder can be adjusted manually.
o Please ask the light microscopy technician for other stage holders if you have a specimen that is not
prepared on a slide
If using the 10x objective start it at the lowest (bottom) position. For the 40x oil, 63x oil, and
100x oil objectives raise the objective (by moving the coarse focus knob clockwise) until the
oil just touches your slide.
8. Focus while viewing through the oculars
Turn on light with the closest buttons to you on the right side
of the microscope
o Mercury (fluorescence)-Top (FL on/off)
o Halogen (bright-field)-Bottom (HAL on/off)
Focus on your specimen and turn the light off.
9. Set up/Select your Dye Configurations
Open the Imaging Setup drop menu from the Setup Manager
list. Check the Show all box on the right side of the menu.
o You may either choose to set up your own track from
scratch using the Imaging Setup and Light Path drop
menus(See Setting Up a New Configuration guide)
o Or you can load a previously saved track by opening
the desired configuration and selecting Apply
Check the configuration set up to make sure you will be able
to see the desired dyes by opening the Light Path drop menu
from the Setup Manager list to see which filters and dichroics
each track is using.
o Note: The light path is highlighted by the software. In
this picture the light is being directed towards ChS
with no light going to Ch2 or Ch3.
10. Choose Image Parameters
Under the Online Acquisition list open the Acquisition Mode
drop box and check the Show all box on the right side of the
o The objective should show the objective you’ve
o Frame size indicates how many pixels in each
direction you want your image to hold
 For publication images we recommend using
1024 x 1024
o Select desired speed
o Averaging
 Number = how many times the image will be
scanned. By increasing this number you can
clean up noisy images
o Under the Scan Area drop menu you’ll find
information on zoom.
 Moves image along X plane
 Moves image along Y plane
 Allows a rotation of the zoomed image
 Zoom magnification
 Each scroll bar has a zeroing button after it to
reset the position back to the original starting
point. There is also a Reset All button beneath
the Zoom magnification scroll bar.
11. Get Image on Computer Screen by selecting the Live or Continuous button. Both will scan continuously until you
select Stop
Live-Uses fastest parameters possible, not the ones you chose in the Acquisition
Mode tab.
Continuous-Uses the parameters you selected in the Acquisition Mode tab.
12. Adjusting PMTs
Under the Online Acquisition list open the Channels drop box
and check the Show all box on the right side of the menu.
o You can adjust the laser intensity with the slide bar for
each indicated laser
o Pinhole-typically start at 1Airy Unit. You can easily
select this by pressing the 1 AU button
o Gain (Master)-Adjusts both the foreground and the
o Digital Offset-Adjusts the background
 Note: If you remove too much background you
will start to remove the foreground image as
o Digital Gain-Adjusts the amplification value of the A/D
To check your pixel saturation
o Check the Show all box on the Dimensions tab located below your image
o Select the Range Indicator option from the Channels drop menu (or click directly on the square indicating
the color of your image)
 Over saturated pixels will be red
 Under saturated pixels will be blue
o Adjust saturation with the PMT slide bars to the desired range.
 Note: You must either select Live or Continuous to see the changes as you make them.
13. Final Image – Click the
button located near the top of the Acquisition tab.
14. Saving your Image
 Click Save from the Open Images toolbar on the right side of
the program window.
o All images will be saved in the D: drive under a folder
with your name on it.
o If you do not have a folder, create one.
15. Shut Down – If someone is signed up to use the scope within two hours after you are finished, leave the Argon laser
on STANDBY and log-off – leaving everything else ON. If you are the last person using the microscope, exit the software
(remember to shut the lasers OFF) and follow the shut-down procedure.
3 – Click Start on the bottom left hand side of the computer and choose the Shut Down option. Wait until the
screens go black and the fan for the Argon laser shuts off before proceeding to 2.
2 – Main Switch (left of microscope)
1 – Mercury Lamp (underneath microscope and to the left)
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