Instrument Name:

Leica Two-photon Upright Microscope (Stellaris 8 DIVE)  Room: X207

Introduction

Combining laser scanning confocal microscopy and two-photon excitation, Stellaris 8 DIVE uses an infrared femtosecond laser as the light source to penetrate deep into tissues and nonlinearly excite fluorescent molecules. The long wavelength reduces laser-induced damage and phototoxicity while ensuring high spatial resolution (imaging depth > 1 mm) and supporting long-term imaging of live samples. The 4TUNE spectral detector freely adjusts the detection band within 380-800 nm and, together with a tunable spectral laser, enables multi-color imaging. The detector features photon counting for fluorescence lifetime imaging; the TauSense module separates signals by lifetime information, adding another dimension to the sample. Single-photon imaging is also supported, with functions essentially identical to the Leica SP8 platform.

Application

The system's high imaging depth and low phototoxicity are suitable for long-term real-time imaging of live animals; the principle that two-photon excitation only stimulates the focal plane allows more precise photoactivation and photobleaching experiments; free spectral adjustment and tunable-wavelength lasers support multi-label sample imaging. Equipped with the TauSense module, fluorescence lifetime imaging can be performed based on dye lifetime properties.

Key Technical Specifications

Light Source:

Continuous-wave laser: 405nm, 488nm, 561nm, 638nm; Femtosecond pulse laser: 1045nm, 680-1300nm adjustable

Imaging Modes:

Wide-field fluorescence, bright-field, single-photon confocal, and two-photon imaging

Detectors:

Built-in (single-photon): 3 spectral HyD S; External (two-photon): 2 spectral HyD NDD (ultra-sensitive), 1 spectral PMT

Objectives:

HC PL APO 10x (Air, NA=0.4, WD=2.8mm); HC PL APO 20x (Air, NA=0.75, WD=0.62mm); HC IRAPO L 25x (Water, NA=1.0, motCORR, WD=2.6mm); HC PL IRAPO 40x (Water, NA=1.1, CORR, WD=0.65mm, salt water available); HC PL APO 63x (Oil CS2, NA=1.4, WD=0.14mm)

Stage:

Motorized stage, movement accuracy 0.1 µm

Scan Resolution:

4×1 to 8192×8192; each channel reaches 8192×8192 with 16-bit grayscale depth

Scan Mode:

High-resolution (Galvano-mirror) and high-speed (Resonant-scanner) modes; high-res 10 fps@512×512, FOV 22 mm; high-speed 28 fps@512×512

Scan Zoom:

0.75x ~ 48x, continuous

Image Processing Software:

LAS X, including 3D visualization, intracellular structure measurement, crosstalk separation, whole-specimen navigation and stitching