Principle of hyperspectral imaging (HSI) as a combination of conventional image processing and classical spectroscopy.
(A) Hyperspectral data cube includes a set of data that are layered on top of one another. Each pixel in the cube consists of an entire spectrum and the resulting image represents the corresponding wavelength band. Typically in hyperspectral imagery, the spatial information is collected in the X-Y plane and spectral information represented in the Z-direction . (B) HSI detector attached to side camera port of an inverted microscope. This combination comprises the imaging spectrograph with sensitive monochrome camera. The CCD camera attached to the spectrograph and the main body through a port. It records the spectral images in a time interval of microsecond-scale. The fluorescence light source is located in the back of camera segment and is adjustable to give an ample light intensity. The front DSLR camera obtains real images of the sample that is located on the heating stage. The filter shield contains combination of filters to detect different fluorescent probes with different excitation/emission rate. (C) Different spectral responses at distinct positions within the detection line. The monocyte is placed within the object field. The multiple fluorescence spectra of cell, conidium or their surroundings are simultaneously captured. The schematic shows a line of areas, and each line covers more than one pixel.