LEICA TCS SP2 AOBS
The Leica TCS SP2 AOBS spectral confocal microscope
Confocal system: The confocal system consists of a Leica DM6000B upright microscope and a Leica DMIRE2 inverted microscope.
Microscopes: The upright stand is used for applications involving slide-mounted specimens; whereas the inverted stand, which has a 37 oC incubation chamber and CO2 enrichment accessories, provides for live cell applications. The microscopes share an exchangeable laser scan head and, in addition to their own electromotor-driven stages, a galvanometer-driven high precision Z-stage which facilitates rapid imaging in the focal (Z) plane. In addition to epifluorescence, the microscopes support a variety of transmitted light contrast methods including brightfield, polarising light and differential interference contrast, and are equipped with 5x, 20x, 40x, 63x (oil and dry) and 100x (oil) Leica objective lenses.
Laser scanning and detection system: The TCS SP2 AOBS confocal system is supplied with four lasers (one diode, one argon and two helium neon lasers) thus allowing excitation of a broad range of fluorochromes within the UV, visible and far red ranges of the electromagnetic spectrum (see table below). The design of the laser scan head, which incorporates acousto-optical tunable filters (AOTF), an acousto-optical beam splitter (AOBS) and four prism spectrophotometer detectors, permits simultaneous excitation and detection of three fluorochromes. The upright microscope also has a transmission light detector making it possible to overlay a transmitted light image upon a fluorescence recording.
Leica TCS SP2 AOBS laser lines
| Laser | Excitation Lines | Suitable Flourescent Dyes |
|---|---|---|
| Diode 20mW | 405nm | Alexa 405, DAPI, Hoechst, Fura 2 |
| Ar 100mW | 458nm | Alexa 458, CFP |
| 476nm | GFP | |
| 488nm | Alexa 488, FITC, GFP, Cy2, DiO, Fluo-3 | |
| 514nm | Alexa 514, YFP | |
| HeNe 1.5mW | 543nm | Alexa 546, TRITC, Cy3, DiI, propidium iodide |
| HeNe 10mW | 633nm | Alexa 633, Cy5 |
The confocal is controlled with dual monitors and running Leica Confocal Software.
Leica Confocal software: The confocal is controlled via a standard Pentium PC equipped with dual monitors and running Leica Confocal Software.
The Leica Confocal Software provides a user-friendly interface for multi-dimensional image series acquisition, processing and analysis, that includes 3D reconstruction and measurement, physiological recording and analysis, time-lapse, fluorochrome co-localisation, photo-bleaching techniques such as FRAP and FRET, spectral unmixing and multicolour restoration.
A ‘Lite’ version of the Leica Confocal Software can be downloaded free of charge from the Leica website. This version of the software is fairly limited in its application but it does allow you to view your image files, make simple volume projections and animations, and perform basic image processing and analysis. To download, please use the link in related links section.
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