Leica DMi8 Manuel utilisateur
2
Table of Contents
3…. Start-up
5…. Microscope Display
6…. Channel Set-up
8… Apply Previous Settings
9…. Display Window
10…. Acquisition Window
11…. Navigator
12…. Objective Resolution Info
13…. Camera Binning Format Info/ Shutdown
3
DO NOT TOUCH ANY BUTTON/SWITCH UNLESS IT IS SPECIFIED BELOW
Start-up
1) Turn on the power strip, It is located on the wall above the power bench.
2) Wait until the display screen at the front of the microscope stand finishes startup
3) If Computer is not ON, turn on the computer.
4) Log in to your lab account on the computer:
Login to the computer using the lab account and password you were given during your training.
Remember to log off when you are finished with your session.
5) Next, open the software “LAS X”
4
A dialog window will pop up. Make sure the selection is the same as picture below, click the
button: OK.
As the hardware configures, you will hear the microscope parts starting to activate. This is
normal. If any error message shows up, take a picture and notice Lai Ding or Teri Zgoda
immediately.
Select your camera: click “Preferences” under camera section.
Then choose the camera under “Set Camera to” section. “Leica DFC9000GT LEFT” is for
fluorescence imaging. “DMC5400 RIGHT” is for bright field color imaging
You are now ready to image! Please save the files to E: drive “User Images” folder and the
corresponding lab folder. DO NOT SAVE IMAGES UNDER C: or D: DRIVE!
5
Microscope Display
Almost all these features can also be changed using the LAS X software.
Status of current selections. Can also change from TL to IL and open/close shutter
Change light type and filter cube
Change the objectives and light path (directed towards eyepiece, left camera port,
or right camera port)
Press to bring the objective all the way down, and to set the focus
6
Channel Set-up
•To add a channel, press the PLUS button in the middle window. To delete a channel press
the MINUS button
•To change the false color of the channel, double click on the gradient rectangle
•Determine what wavelengths you need. Select the filter cube, which will be outlined in
red when active.
•Turn on the LED source. You may have to change the wavelength it is emitting at using
the dropdown below the slider. If it can’t use that wavelength with that filter, the box will
be grayed out with a line through it.
Make sure that the wavelengths in these dropdown boxes are the same throughout all
the channels. This quickens the time it takes to take a photograph.
•Use the slider to increase/ decrease the light intensity
•Press the LIVE button in the lower left-hand corner to view your subject
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•Use the knob circled in red to adjust Z
•Use the knobs bellow to adjust X and Y. There is a small toggle switch circled in red below
that changes between fine and coarse movement.
•Adjust exposure time and laser intensity for each channel.
•Under PROJECT SETTINGS dropdown, uncheck SEQUENCER MODE
•Press SINGLE IMAGE to photograph one single channel.
•Press CAPTURE IMAGE to photograph all channels.
•Press START to begin a z-stack or time series.
8
Apply Previous Settings
•Open a previous project using button circled in yellow
•Go to “Configuration” tab and “Param Recall” section, click “Load” button, under the
pop-up file dialog window, select “setting.xml” file
•Go back to “Acquire” tab, press the button circled in red to apply the settings
9
Display Window
d
Annotation: Add shapes or scale bars to image. Measure distance between points.
Select what
channels you want
to view.
This button is to
view a composite of
all the channels.
Open 3D viewer
window. Useful only
if you’ve taken a Z-
stack.
•Change Look Up
Table (color,
intensity, or black
and white)
•Autoscale all
channels
•Stack autoscale
•Channel auto scale
•Reset
•Use background LUT
to scale
•Open Histogram
Look up table and contrast. Moving the two circles closer together will increases contrast.
10
Acquisition Window
Select Zfor Z-stack and Tfor time series.
Mark and Find
Tile Scan
FIM: fluorescence intensity manager, higher % more light
IL –FLD: field stop, should always be at 6
Camera %: (light is directed towards eyepiece) or 100 (light is
directed to camera)
Change objective here
Keep “Magnification Changer” as “ 1 “
Binning: select if you want a faster, quicker image with less
resolution exposure here.
Longer exposure = more light = higher chance of photobleaching
Binning: increases exposure but decreases resolution
Scroll through the Z to find when your subject just begins to dim and
go out of focus, then press BEGIN. Go through the focus to the next
point the subject starts to go out of focus and click END.
The system will provide number of steps with “optimized” Z-step
size if “System Optimized” box is checked. User can assign step size
or number of steps with corresponding box checked.
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