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LEICA TCS SP2 AOBS

The Leica TCS SP2 AOBS spectral confocal microscope.

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.

The confocal system consists of a Leica DM6000B upright microscope and a Leica DMIRE2 inverted microscope.

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

    Leica TCS SP2 AOBS laser lines.
LaserExcitation LinesSuitable Flourescent Dyes
Diode 20mW405nmAlexa 405, DAPI, Hoechst, Fura 2
Ar 100mW458nmAlexa 458, CFP
 476nmGFP
 488nmAlexa 488, FITC, GFP, Cy2, DiO, Fluo-3
 514nmAlexa 514, YFP
HeNe 1.5mW543nmAlexa 546, TRITC, Cy3, DiI, propidium iodide
HeNe 10mW633nmAlexa 633, Cy5

photograph © Marc Isaacs

The confocal is controlled with dual monitors and running Leica Confocal Software.

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.

Leica Confocal Software showing control and display screens.

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.

 

Page content written and maintained by Dr. A.J. Hayes and Mr. M. Isaacs

For further information:

Dr Anthony J. Hayes

Position:Facility Supervisor & Administrator, Confocal Microscopy
Telephone: +44(0)29 208 76611Extension: 76611

Mr Marc Isaacs

Position:Histology and Imaging Technician
Telephone: +44(0)29 208 75139; 208 76611Extension: 75139/76611

Dr Elizabeth Gealy

Position:Technician (Histology & Confocal Unit)
Telephone: +44 (0) 2920 875139Extension: 75139
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