CBIS Confocal: TCS SP5 X

TCS SP5 X with two photon excitation laser

Leica

The Leica TCS SP5X Confocal Micro-scope System with multi-photon laser (MP) consists of an inverted microscope DMI 6000 CS with a XY scanning stage, Z motorized focus as well as a Super-z galvo stage, for higher speed and repeatability. It uses an Acousto Optical Tunable Filter (AOTF) for laser line selection and intensity control with micrscosecond precision. 

LocationCBIS Confocal Microscopy Laboratory
Booked CBIS equipment before? Information for First Time Users

About the Leica TCS SP5 X

An Acousto Optical Beam Splitter (AOBS®) replaces fluorescent filters and beam splitters in the light path. It is a spectrally tunable element which is fast, switchable and without any moving parts. In combination with AOTF, the AOBS offers a more light efficient system. This optical element allows the excitation light to excite the sample and the emission passes straight through the crystal.

The Spectral Detector SP ® is free from barrier filters and secondary dichroic filters and therefore highly transparent and extremely sensitive. Light emitted from the sample is dispersed by a high efficient prism. The resultant spectrum is directed towards two tunable slits. Two motorized sliders acts as barriers and transmit a step-lessly tunable band to the sensor. The full spectrum can be split into several channels without any loss of energy. The Spectral Sliders offer a flexible system for wavelength discrimination which can be used to minimize cross talk. The SP5 has also a Tandem Scanner which comprises of a conventional scanner for high resolution morphology imaging (8kx8k) and a resonant scanner for fast imaging. Conventional speed ranges from 10-1400Hz whereby resonant speed is 8000Hz.

For laser excitation, the user has a choice of different lasers: a diode 405, a multiline Argon, a White Light Laser (WLL) and a Ti-sapphire IR laser. With white laser light, users can freely choose any excitation line within the continuous range of 470nm to 670nm in 1nm increments to perfectly match fluorophores with minimum cross-excitation, and turn down laser power to prevent specimen damage. There are up to eight excitation lines can be used simultaneously. The multiphoton excitation allows imaging of fluorophores deep into intact tissue (beyond the range of confocal microscopy). This penetration is due to the greatly reduced scattering of infrared light and the fact that excitation is restricted to the focal point of the pulsed laser.

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  • Features
    • Leica DMI 6000 CS Bino Inverted Research Microscope with motorized xy stage and SuperZ-Galvo Stage
    • Fluorescent illumination: Leica EL6000
    • Filter system: BGR filter system: UV, BP420/30, BP465/20; Blau, BP495/15, BP530/30,Grun, BP570/20, BP640/40
    Objectives
    • Leica HC PL APO 10x/0.40 CS
    • Leica HC PL APO 20x/0.70 lmm Corr CS
    • Leica HCX PL APO 40x/0.85 Corr
    • Leica HCX PL APO 63x/1.20 W CORR CS
    • Leica HC PL APO 100x / 1.40 Oil CS2
    Lasers
    • Diode laser, 405nm 50mW
    • Argon Ion Laser: 458nm 5mW, 476nm 5mW, 488nm 20mW, 496nm 20mW, 514nm 20mW
    • White laser: 470nm to 670nm, with 1 nm increments, with 8 channels AOTF for simultaneous control of 8 laser lines, each excitation wavelength provides 1.5mV
    • Multi Photon: Chameleon Ultra II, Ti-Sapphire, tuning range 680-1080 nm, ~4W at 800nm
    Detectors
    • One Channel TLD Bright field Detector
    • Three Multi-Alkali PMT detectors
    • Two non-descanned detectors (NDD) for the multi-photon with filter block for FITC / TRITC
    Software
    • Leica Application Suite Advanced Fluorescence (LAS AF)
    Applications
    • High Resolution imaging
    • High Speed imaging
    • Spectral imaging
    • Advanced F-techniques with wizards: FRAP, FRET AB/SE
    • Excitation lambda scans
    • Photo activation / ablation experiments