Optimal selection of flurophores for bioimaging using hyperspectral cameras
DOMAIN In many situations it would be beneficial to have an instrument that can show In biology and medicine, fluorescent labelling is used to detect the location and concentration of various molecules. This can be used to discover how molecules are interacting in relation to diseases and biological functions. Often, it is of interest to study the distribution of multiple marker molecules in a tissue simultaneously. This, however, is made difficult by the fact that most fluorophore labelling methods are based on single wavelength channel detection which in creases the likelihood of spectral overlap of the fluor- ophores.
DESCRIPTION To allow for multiple fluorophore molecules in bioimaging, a camera which detects a whole spectrum in each pixel is necessary. Such hyperspectral cameras produce image cubes where each pixel contains not only intensities from the red, green and blue channel as in colour photos, but intensities from hundreds or thousands of wavelengths. The use of chemometric met hods enables the resolving of the mixed spectra into pure component estimates such that the concentrations of the different fluorophores (and background species) can be estimated. However, the quality of the concentra- tion predictions is dependent on variables such as the number of fluorophores, the degree of spectral overlap and the noise level. So, in order to obtain the best results the current invention describes a mathematical method for selecting an optimal set of fluorophores for use in bioimaging problems. The method takes also into consideration the set of background spectra that is also present in most experiments.
UNIQUE SELLING POINTS
• Large number of fluorophores can be detected simultaneously
• High level of signal to noise ratio can be achieved
• Scanning of micro-array plates to enable multiple samples on a single slide
• Use in immunohistochemistry
STATE OF DEVELOPMENT A hyperspectral microarray scanner has been built and is ready for testing of the main method.
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