Leica TCS SP8 User manual

Leica TCS SP8
Quick Start Guide

Leica TCS SP8 System Overview

Start-Up Procedure
1. Turn on the CTR Control Box, EL6000 fluorescent light source for the
microscope stand.
2. Turn on the Scanner Power (1) on the front of the Compact Supply Unit (CSU)
3. Turn on the Laser Power (2)
4. Enable the lasers by turning the key
from the Off position to the On
position (3).
5. Turn on Workstation and Monitor
6. Log into user profile
7. Double click on the LAS X icon to start the software.
8. Select Machine for the Configuration and DMi8 for the Microscope. (the
simulator is used to view and analyze images and not acquistition)
10. Click OK to start the initialization of
the LAS AF (Leica Application Suite
for Advanced Fluorescence)
11. A message will appear asking whether you want to initialize the stage. Initializing
the stage is required to activate the Tiling and Mark & Find features in the software.
If you select yes, the stage is going to
move to all 4 corners to calibrate the
position of the stage.
Make sure there is nothing on the
stage that will hit the condenser
before you select yes!

Shut-Down Procedure
1. Turn off the lasers in the software
2. Lower the objective to the lowest position, remove specimen and clean all of the
objectives
3. Save all unsaved data and exit LAS X.
NOTE: Do not turn off scanner and/or CTR control box for the microscope before the
software is closed
4. Turn off the switches on the front of the laser supply unit in the reverse order (#3-#1)
of start-up (ie. turn key off, then turn off Laser Power and then turn off Scanner
Power)
5. Turn off microscope control box

The LAS X software will open in the Acquire tab. There are 3 portions to this window:
Scan Parameters Light Path Image

Turning on the Lasers:
1. Click on the “+” in the Laser Lines Box in the Beam Path Settings Window to directly
open the main Laser Control Window
2. Activate the lasers you require
3. Alternatively, click on the Configuration tab and the Laser icon to open the main
Laser Control Window
NOTE: Only turn on the laser(s) with the appropriate laser line(s) that will excite
the fluorophores you are using

Additional Tools in the Configuration Tab…
1. Customize the USB Control Panel -assign various parameter and sensitivities to
the various knobs of the control panel
2. Specifications of the Objectives equipped on the microscope
Save and Load the customized settings
Alternatively , click on the Control
Panel Icon in the Beam Path Settings
Window as a short cut to the USB
Control Panel Window

Additional Tools in the Configuration Tab Continued…
3. Dye Database with the excitation and emission spectra of common fluorochromes
Emission spectrum from a lambda scan can be added to the dye database.
Many manufacturer’s will also provide the data for fluorophores that can added to
the database (see LAS AF Help)
Another great resource is the Leica FluoScout
interactive tool
http://www.leica-microsystems.com/fluoscout/

Beam Path Setting
Option #1: Manual setting of the beam path configuration
1. Click On to activate
the lasers 2. Adjust the laser intensity
of the appropriate laser
line(s) by moving the
slider up or by directly
entering the level (start
low as a suggestion).
4. Use Autoselect to select the beam splitter
(inactivate and manually select if BSC is red)
3. Select the appropriate objective
6. Select the appropriate emission
spectrum (if available) from the
dye data base to use as a guide
8. Define the of emission to be collected with the sliders
5. Click On to activate the
appropriate detector(s)
Keep in mind that the emission spectra
displayed are to be used as a guide and are not
defining the wavelengths collected. The
specific wavelengths collected are determined
by the position of the gates.
Alternatively, double click on
the slider to open a window
that allows you to directly
enter the start and end
position of the gates.
7. Select the Pseudocolour