ZEISS AXIOVERT 200M LSM 510 META CONFOCAL MICROSCOPE

This is a top of the range confocal microscopy system, versatile and fully equipped for a multitude of specialised tasks, as required for a true multi-user core facility.
The system is designed in a modular way, allowing for hard- and software upgrades at a later stage - including the addition of a pulsed infrared laser for 2-photon excitation and imaging (NLO).

Summary of functionality:
In addition to the well-known general benefits of confocal microscopy, e.g. the ability to scan and clearly image tissue specimens of increased thickness (up to ~100 µm, depending on tissue type and labelling) and to perform optical sectioning, Z-projections and 3D reconstructions of scanned specimens, this system is currently equipped to perform the following:

Multi-channel fluorescence imaging: The system is equipped with Argon laser lines (excitation at 458, 477, 488 and 514 nm; blue range), a solid state laser (561 nm; green range) and a HeNe laser (633 nm; red range). It is thus possible to use most commonly available fluorescent dyes and fluorescent proteins simultaneously for labelling experiments. There are two conventional confocal detectors, plus a polychromatic, spectral detection channel (META-detector) allowing simultaneous detection and separation of up to 8 emission spectra. In addition, the system has a confocal detector for transmitted light, allowing optical sectioning of translucent, unlabelled specimens or in combination with fluorescence labelling.

The system is not equipped with an ultraviolet laser (this would not be compatible with the NLO configuration and has many disadvantages such as a high degree of phototoxicity); however, it is possible to use the conventional HBO light source for confocal visualisation of e.g. DAPI-labelled cell nuclei.

Multitracking, spectral detection, emission fingerprinting and channel unmixing:
This provides for clear separation and quantification of all fluorochromes used (eliminates "bleed-through"-problem) and allows to distinguish actual signals from autofluorescence. Moreover, many different fluorophores with closely overlapping spectral ranges (e.g. varieties of fluorescent proteins) may be used simultaneously.

Selection of regions of interest (ROIs or spots) for scanning, measurements and experimental manipulations:
Free selection of ROIs in x, y and z direction permits image acquisition, physiological measurements (e.g. ion-sensitive dyes and ratio imaging) and experimental manipulations, such as Fluorescence Recovery after Photobleaching (FRAP), fluorescence Loss in Photobleaching (FLIP), fluorescence Resonance Energy Transfer (FRET), photo-uncaging or photo-activation of probes in defined regions, down to sub-cellular dimensions.

Integrated incubator unit:
For observation of live cells and tissues in a controlled atmosphere; combined with the x-y-z motorised microscope stage and a choice of confocal or conventional detection using a CCD camera, this permits efficient live cell observations and time-lapse recordings, e.g. in cell migration studies.

Specialised application software modules:
These include applications for physiological measurements (e.g. ion concentrations), FRET and FRAP. Advanced image processing, analysis (e.g. colocalisation measurement) and database functionality is provided by the Zeiss LSM core software, and an off-line version of the software will be available for post-acquisition image processing.
A visual macro editor is provided to assist in the design of complex experimental, data acquisition and analysis routines.
Specialised stereology software dedicated to morphological analysis of cells and tissues (particularly nervous tissues) will be provided by a third-party supplier.

Manuals for the Confocal Microscope >>>

	LSM 510	(1 Mb)
	LSM Guided Tour	(4 Mb)

ZEISS FLUORESCENCE MICROSCOPES

1. The Zeiss Axiovert 200 is a sophisticated inverted motorised fluorescence microscope with phase and DIC contrast, 10x, 20x, 40x dry and 100x oil immersion objectives. The following laser lines are available e.g. Dapi, Alexa, Cy3, GFP and Cy5. It is also equipped with a monochrome Zeiss Axiocam HR and Axiovision 4.6 software.
   
   
2. The Zeiss Axioskop 2 upright motorised fluorescence microscope with bright field, dark field, phase and DIC contrast with 5x, 10x, 20x, 40x dry and 100x oil immersion objectives. Three incident light filters are available, blue-violet, the violet and green. The microscope is equipped with a colour AxioCam HR and Axiovision 4 supporting software. This microscope is the property of the Department of Medicine.

Zeiss website:  http://www.zeiss.com

LIGHT MICROSCOPES

1. The Nikon light microscope with phase and DIC contrast, 4x, 10x, 20x dry, 40x and 100x oil lenses are available with a colour digital camera and software.
   
   
2. A Nikon two header microscope is also available.
   
   
3. A Nikon 5 header microscope for teaching purposes is also available.
   
   

COMPUTER WORKSTATIONS

Two workstations will be available with the LSM Manual, quick start and guided, an off-line version of the LSM software for post-acquisition image processing, Axiovision 4.7, Photoshop and morphometric and stereology programmes, Microsoft Office and Nero.