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CIAN Leica SP8 – short instructions, version 3.1, September 2015
SHORT INSTRUCTIONS FOR OPERATING Leica SP8 AT CIAN
1 - Equipment Setup
Fig 1: Basic components of the Leica SP8 microscope
1: Leica DMI 6000B inverted microscope
2: Scan head
3: Anti-vibration table
4: EL6000 metal halide lamp for WFF
5: CTR control box for microscope stand
6: Smart Move joystick
7: Monitor
Fig 2: Spectral detection system, schematic
Prism
Emission
Light
Version 3.1, September 2015
8: Control panel
9: Keyboard
10: Computer mouse
11: Computer table
12: Lasers in Compact Supply Unit (CSU)
13: Supply Control
14: Workstation
HyD
3
1
PMT
PMT
4
2
PMT
Sliders
page 1 of 10
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Summary of Contents for Leica SP8 AT CIAN
- Page 1 CIAN Leica SP8 – short instructions, version 3.1, September 2015 page 1 of 10 SHORT INSTRUCTIONS FOR OPERATING Leica SP8 AT CIAN Version 3.1, September 2015 1 - Equipment Setup Fig 1: Basic components of the Leica SP8 microscope 1: Leica DMI 6000B inverted microscope...
- Page 2 CIAN Leica SP8 – short instructions, version 3.1, September 2015 page 2 of 10 2 - Starting up Leica SP8 Note: Solid state lasers and the EL6000 widefield light source require only minimal warm up, but adhere to 15min/15min rule: leave on at least 15min before shutting off, leave off at least 15min before restarting 1.
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Page 3: Cleaning Objectives
CIAN Leica SP8 – short instructions, version 3.1, September 2015 page 3 of 10 2.2 - Cleaning objectives Clean immersion objectives before and after use. The dry objectives on the SP8 should not be cleaned in this fashion since they have concave front lenses. - Page 4 CIAN Leica SP8 – short instructions, version 3.1, September 2015 page 4 of 10 3.2 - Using LAS X for microscope control and image acquisition Open the LAS X software as described in section 2. 3.2.1 - LAS X main window, general functions The main window opens in the Acquire mode.
- Page 5 CIAN Leica SP8 – short instructions, version 3.1, September 2015 page 5 of 10 3.2.4 - Beam path setting a) Single wavelength or simultaneous acquisition 1. Click ON to activate the lasers (should already be on, see 3.2.2). 2. Adjust the laser intensity of the...
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Page 6: Acquisition Parameters
CIAN Leica SP8 – short instructions, version 3.1, September 2015 page 6 of 10 3.2.5 - Acquisition Parameters Format, or number of pixels in the scan area; for • optimal format adjust to desired pixel size, see below Scan Speed; start with 400 to 600 Hz to avoid •... - Page 7 CIAN Leica SP8 – short instructions, version 3.1, September 2015 page 7 of 10 3.2.7 - Acquiring Z stacks 1. Select xyz Acquisition Mode. 2. a) For absolute stack limits: • In live scan, use the Control Panel (stage Z) to move the focus to one extreme of the stack.
- Page 8 CIAN Leica SP8 – short instructions, version 3.1, September 2015 page 8 of 10 3.2.8 - Additional Acquisition Modes – Mark & Find and Tiling The stage must be initialized during startup for this to work. Careful: Clicking in the stage area schematic will move the stage to the clicked position.
- Page 9 CIAN Leica SP8 – short instructions, version 3.1, September 2015 page 9 of 10 4 - Shutting down Leica SP8 1. Turn off the lasers in the software. 2. Lower the objective to the lowest position, remove specimen and clean all of the objectives that were used in this session;...
- Page 10 Appendix: Technical data for Leica SP8 Microscope: Leica SP8 point-scanning confocal system, on a Leica DMI6000B inverted microscope, fully motorized, with Leica “SuperZ” Galvo stage, four spectral fluorescent light detectors (three PMT, one HyD high sensitivity detector), one transmitted light detector (PMT) Objectives:...
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