Laserový skenovací mikroskop Leica Stellaris 8 s Falcon FLIM

Detailed description

Methods

Confocal Scanning Fluorescence Imaging: Equipped with the Lightning super-resolution module for enhanced imaging precision.

Transmitted Light Imaging: Supports brightfield and Nomarski contrast (DIC), fully automated.

Tile-Scanning & Image Merging: Enabled through LAS X software for seamless large-area imaging.

Dynamic Experiments: Includes time-lapse and multi-position imaging capabilities.

Photo-Kinetic Studies: Suitable for experiments like photobleaching (FRAP) and activation.

FRET Analysis: Covers both Sensitized Emission (SE) and Acceptor Photobleaching methods.

Advanced Techniques: Includes FLIM and FRET-FLIM for in-depth fluorescence lifetime and interaction studies.

Lasers

405 nm cw diode laser

488 nm cw FRAP laser

440 – 790 nm pulsed white light laser (WLL)

• laser wavelength and intensity are regulated by the integrated AOTF and AOBS
• wavelength selection is adjustable with 1 nm precision
• enables up to 8 independent parallel laser lines
• possible laser frequencies: 80, 40, 20, 10, 5, 2.5 MHz

2x HyD S - most sensitive detector at 410 – 560 nm

2x HyD X - most sensitive detector at 560 – 740 nm. The fast detector is optimised for FLIM, and the image quality is better with a resonant scanner than HyD S.

HyD R - most sensitive detector above 740 nm. The fast detector is optimised for FLIM, and the image quality is better with a resonant scanner than HyD S.

T-PMT – transmitted light detector

Transmitted light techniques

Brightfield and Nomarski contrast (DIC) (automated)

Objectives

For the current setting check the table below

Fluorescence light source

HXP 120W/45C VIS Hg lamp – Leica EL6000

Filters for fluorescence

DAPI (Ex. 355/56; DC: 400; Em. 460/50)

DA/FI/TX (Ex. 403/18 + 493/18 + 575/25; DC: 435; 505; 595; Em. 456/30; 530/35; 630/60)

GFP (Ex. 470/40; DC: 495; Em. 525/50)

Y5 (Ex. 620/50; DC: 660; Em. 700/75)

Live cell imaging

OKOlab incubation system with integrated regulation of CO₂, O₂, temperature, and humidity

Software

LAS-X

Modules (fully implemented to the LAS X software):

• LAS X LiveDataMode
• LAS X 3D Visualisation
• LAS X Phasor
• LAS X FRAP
• LAS X Assay (Navigator)
• LAS X Lightning (computational super-resolution module)