Leica SP8-STED (Inverted)

Learn more about Stimulated emission depletion (STED) microscopy:

STED Confocal Super-Resolution - Leica TCS SP8 STED 3X

Microscopy: Super-Resolution: Overview and Stimulated Emission Depletion (STED) (Stefan Hell)

The Guide to STED Sample Preparation

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

• Which objectives are installed in this system?

You can find the list at the end of this page, file “Objectivefinder_ STED.pdf“.

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

Find your fluorophore on FPbase.

• 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

Or you can work on 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.

• Can I do Fluorescence Lifetime Imaging Microscopy (FLIM)?

What is FLIM - Fluorescence Lifetime Imaging?

And yes, you can do FLIM on the Leica SP8 microscope, which is equipped with hybrid photon-counting detectors (HyD).

• How to adjust the motorized collar correction ring on the 86X and 93X objectives?

Follow this guide.

Offers resolution up to 30nm lateral and ~100nm axial in 3D. A combination of gated, spectral detection, and white light laser provides an excellent signal with full freedom in fluorophores' choice at the visible range.

Our system:

1. Lasers:

   1. 405

   2. Argon

   3. White Light Laser (WLL)- adjustable excitation

   4. STED dep lasers (594,660,775)

2. Objectives:

   1. 10x/0.4 air

   2. 20x/0.75 air

   3. 40x/1.1w

   4. 63X/1.2w

   5. 63/1.4o

   6. 86x/1.2 w (STED)

   7. 93X/1.3 glycerol  (STED)

   8. 100X/1.4 o (STED)

3. 3HyD Detectors , 2PMTs

4. Scanners- galvo/resonant

5. Falcon -Lifetime imaging