Leica SP8 confocal-2Photon

Learn more about Leica SP8 2Photon:

Multiphoton Microscope Leica TCS SP8 MP

Two-Photon Microscopy

Cameras and Photosensitive Detectors I: How Do They Work?

Galvo v Resonant scanning guide

Opening .lif files in FIJI

Frequently Asked Questions:

• How do I add and clean immersion on and off objectives correctly?

Proper Use of Objective Immersion Oil

Oiling an Objective

Determining if you’ve used the right amount of oil

Removing oil from the lens

• How do I choose the right excitation and emission settings for my fluorophore?

Find your fluorophore on FPbase. Based on the excitation and emission data, chose the closest filter set in the software.

• How do I save and access my data?

Save the data in:

→ open the driver data (D :) “Data_Bioimg”

→ open folder with your Weizmann user name

→ open a new folder with the date (crucial)

→ save the data (characters like ~!@#$%^&*()`;’:<>,/?[]{}  are not allowed, and files including them are not copied)

Once you are done, the computer will synchronize the correctly saved data to the “bioimg” server.

To see the data on your PC, follow the instructions in this link: BioImg Storage Server system.

**When starting to work with the Bioimaging system for the first time, please make sure you have a UNIX ID added to your WIS ID (you can check it with your unit administrator)

• How can I visualize the data on my pc?

Download LAS X; when you install it, chose “confocal.”

You can open it in Imaris free Viewer

You can open it in ImageJ

You can open it on one of our Image Analysis Software and Workstations

• How do I open Leica files in Fiji and Imaris?

Opening .lif files in FIJI

Follow the instructions here.

Or download macro from here.

• Which objectives are installed in this system?

You can find the list at the end of this page, file “Objectivefinder 2P.pdf“.

Leica TCS SP8 Confocal - MultiPhoton 

Suitable for cell cultures, plants, tissue sections, or thicker samples.

Enables Z compensation for even illumination.

1. Excellent spectral separation and high sensitivity, combining Acuso-Optical-Tunable-Filter, spectral detection, and 1 PMT and 2 HyD internal detectors. 

2. Chameleon Vision II coherent tunable laser, combined with 2 sensitive HyD external detectors (NDD), enables Second-Harmonic-Generation detection in 3D and 4D.

3. Large fields of view acquired in fast scanning mode enable accurate navigation within the sample during high-resolution imaging.

4. Environmental control enables live imaging for longer periods, combined with a fast resonant scanner, sensitive detectors, and software solutions for lower phototoxicity.

5. LASX software pack, including integrated Deconvolution module.

6. Variety of optional sample holders for increased imaging stability of cleared or expanded sample.

Body Configuration - Upright

Lasers -

UV

• 405

Visible

• 458nm

• 476nm

• 488nm

• 496nm

• 514nm

• 561nm

• 594nm

• 633nm

Multiphoton laser -

Chameleon Vision II  - Coherent tunable ultrafast Ti: Sapphire laser

• 680-1080 nm. 

 Scanner

 Tandem scanner 8 kHz

 AOBS

Detectors

Reflected Internal Fully spectral

• 2 X HyD SP GaAsP-Detektor (HyD are characterized with high sensitivity and low noise detection)

• 1 X PMT SP confocal detector

NDD detectors

• Reflected - 2 X  HyD-RLD

• Transmitted detector

Objectives

1. HC PL FLUOTAR 10x/0.30 (air)

2. HC PL APO 20x/0.75 CS2 (air)

3. HC FLUOTAR L 25x/0.95 W VIS (water dipping)

4. HC PL APO 40x/1.10 W CORR (water immersion)

5. HC PL APO 63x/1.40 OIL CS2 (oil immersion)

6.  HC FLUOTAR L 25x/1.00 IMM  Leica motCORR - n_e=1.457 (for cleared samples)