| | SIRF (Simple Internal Reflection Fluorescence) | |  |
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| The SIRF diaphragm slider (not available in the USA and Japan) enables a new contrasting technique for fluorescence applications with the Alpha Plan Fluar 100x/1.45 oil immersion objective. |  back
 Principle
Advantages and Benefits
Components
 Deutsche Version |
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The technique is related to TIRF Microscopy, in which a laser beam illuminates the sample at what is known as the critical angle.
At this angle, the excitation light, rather than being transmitted by the specimen, is totally reflected by the interface between the cover slip and the aqueous medium ( Principle). |
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Using the SIRF slider and the alpha Plan Fluar 100x/1.45 oil immersion objective, the microscopist can selectively excite fluorochromes located very near the cover slip.
Therefore, the technique is well suited to the investigation of processes taking place in or on the cell membrane, or very close to it. |
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Improved contrast and higher resolution with the Carl Zeiss SIRF slider | |
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| SIRF microscopy has the fundamental advantage over TIRF microscopy that it can be used with standard HBO/XBO light sources and, accordingly, with less complex equipment. | |
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MDCK cells were transiently transfected with a GFP construct. The protein is localized in the cell membrane.
Cell preparation: Doris Meder (Laboratory of Kai Simons), Max Planck Institute of Molecular Cell Biology and Genetics, Dresden
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