Abstract
Light emitting diodes (LED), which are available as small monochromatic light sources with characteristic features such as maximum illumination power combined with minimum energy consumption and extremely long lifespan have already proved as a highly potential low-cost alternative for specific diagnostic applications in clinical medicine such as tuberculosis fluorescence microscopy. Likewise, the most reliable evaluation of Her-2/neu (c-erbB2) gene amplification, which has been established in the last few years for routine diagnosis in clinical pathology as determinant towards Herceptin-based treatment of patients with breast cancer, is based on fluorescence in situ hybridization (FISH) and corresponding high priced fluorescence equipment. In order to test the possibility to utilize the advantages of low-cost LED technology on FISH analysis of c-erbB2 gene expression for routine diagnostic purposes, the applicability of a standard bright field Carl Zeiss Axiostar Plus microscope equipped with a Fraen AFTER* LED Fluorescence Microscope Kit for the detection of Her-2/neu gene signals was compared to an advanced Nikon Eclipse 80i fluorescence microscope in combination with a conventional 100W mercury vapor lamp. Both microscopes were fitted with the same Quicam FAST CCD digital camera to unequivocally compare the quality of the captured images. C-erbB2 gene expression was analyzed in 30 different human tissue samples of primary invasive breast cancer, following formalin fixation and subsequent paraffin-embedding. The Her2/neu gene signals (green) were identifiable in the tumor cells in all cases and images of equal quality were captured under almost identical conditions by 480 nm (blue) LED module equipped standard Axiostar microscope as compared to conventional fluorescence microscopy. In this first attempt, these monochromatic LED elements proved in principle to be suitable for the detection of Her-2/neu gene expression by FISH. Thus, our own experiences emphasize the high potential of this technology to provide a serious alternative to conventional fluorescence microscopy in routine pathology; representing a sustainable technological progress, this low-cost technology will clearly give direction also to the growing field of molecular pathology.* AFTER = Amplified Fluorescence by transmitted Excitation of Radiation
Highlights
Light Emitting Diodes (LED) are characterized by low cost, effective energy consumption and extremely long lifespan when compared to conventional light sources [1]
The availability of a variety of small monochromatic Light emitting diodes (LED) modules efficiently emitting the spectrum in a single desired bandwidth has stimulated the interest of clinical researchers to utilize this low-cost technology for advanced fluorescence microscopy in diagnostic research [4]
Since LED based light sources operate without increasing their temperature, common safety problems related to considerable heat production by conventional high pressure mercury lamps are completely avoided
Summary
Since LED based light sources operate without increasing their temperature, common safety problems related to considerable heat production by conventional high pressure mercury lamps are completely avoided. LED modules attached to standard light microscopes have been successfully applied in fluorescence-based screening of tuberculosis, pointing to the considerable reduction of related costs combined with increased safety and, as a consequence, to the potential for low-income countries to perform such advanced diagnostics of this disease in the near future [5,6].
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