Northwest Leica DM6B User manual

Guide for LAS X Navigator (Tile scanning)
1. Initial set-up
~ Place slide on stage ‘coverslip up’.
~ For transmitted light imaging: reference Quick Guide for Bright Field (color) imaging.
~ For fluorescent light imaging: reference Quick Guide for Fluorescent imaging.
~ Focus on sample. Find cell/region of interest.
~ Set imaging settings as required by experiment.
2. Setting up Tile Scanning
~ Press the Navigator button located near the top of the Open
Projects/Acquisition panel. New window will open.
~ In the new window find access to the standard LAS X settings on the left side (A-G). Adjust settings at
any time if required while staying in the Navigator mode.
~ Press “Carrier (G) > Well Plate > 4 slide” to view screen as 4 slide stage.
~ Using mouse wheel, zoom in/zoom out to the field of view of the interest.
~ Double clicking by left button of mouse will preview new areas of the slide. Also, Spiral button (H) can
be activated to preview sample by spiraling pattern. Stop it when needed.
~ Using one of Shape tools (I) draw border round the region(s) of interest (J). Each selection will be listed
in a Task List (K).
Leica DM6B Widefield
Quick Start Guide

~ To create even focus across selected region at image
acquisition step activate Focus Points button (L) and
place 3-6 focal points in different parts of each selected
area. The number of focal points depends on the size of
the selection.
~ Press “Live”, then open Focus Map (M), select the first
point, adjust focus by focus knob and press “Set Z”. Repeat
manually for each focal point, or use “Find all by
Autofocus”. Close Focus Map window (M).
~ Note: pay close attention to autofocus performed by software. If some points are found out of focus, you
can manually adjust it.
~ When all settings are set, click "Start" to start the acquisition.
~ Each selected region will be saved in Projects tab (A) as “Region000_Merged” file.
~ Note: If image had not been merged at acquisition:
a) Go to “Stage tab” (F) and make sure “Merge images
during acquisition” in ON.
b) Manually merge required images. In the Navigator
window go to the “Mosaic Merge” tab, select file to be
merged and press “Merge” button located in the bottom
right corner. Newly created image will appear in the
Projects tab (A) as “Region000_Merged” file.
~ To exit LAS X Navigator window, press “Close” (N). All progress will be saved.
3. Save images
~ In the “Open projects” tab, click “Save All”. Select the appropriate
directory and change the Experiment file name, if desired. Click OK to
save.
4. Shut Down
~ Remove sample. Clean oil off objectives with lens paper if used. Return objective turret to 10x.
~ Transfer files to flash drive or network storage. Close software.
~ If there is another user coming within 1 h, stop here, leaving microscope ON.
~ If not, proceed with following steps for complete shutdown:
~ Turn OFF the microscope controller box (A). Cover microscope.
~ End reservation in NUCore.
If help is needed, please see BIF staff