This resource will develop as questions are asked. Please feel free to submit a question to any of the people involved in the facility.

Why should i use a confocal?

Confocal microscopy allows the rejection of fluorescent light emitted from parts of the sample above and below the plane of focus, using an aperture, referred to as the pinhole, in front of the detector. This gives an increase in the resolution and improved signal to noise ratio in the resultant image of a single plane through the sample. With the inclusion of z-scanning a high quality 3-dimensional view of the sample can be rendered on a computer.

More information:

• Pawley, J.B. (Ed.) 2006. Handbook of Biological Confocal Microscopy (3rd edition) Springer, New York.
• Murphy, D.B. 2001. Fundamentals of Light Microscopy and Electronic Imaging. Wiley-Liss, New York.
• Molecular Expression Microscopy Primer - confocal pages

What is Koehler illumination?

Once set up properly, Koehler illumination (also spelt Köhler), provides and even and bright illumination of the specimen for brightfield microscopy, offering the maximum resolution and contrast in an image. It is achieved by adjusting the field diaphragm, focussing and centering of the condenser and adjusting condenser aperture diaphragm once a sample is in focus.

More information:

• Murphy, D.B. 2001. Fundamentals of Light Microscopy and Electronic Imaging. Wiley-Liss, New York.
• Molecular Expression Microscopy Primer - Köhler illumination pages

What do the acronyms for filters mean?

Unfortunately this depends a bit on the manufacturer. The Nikon MicroscopyU Introduction to Fluorescence Microscopy website will help, both on the theory of fluorescence microscopy and the terminology used.

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