Fig 1.
From the light source (LED), the light line passes the fast filter wheel (F1) and lens (L1) and reaches the samples. The light focuses on and illuminates the samples through an objective (OL). PM is the pulse modulator, which controls the motored stage step by step. The light beam is adjusted through a tube lens (TL) and beam splitter (BS). A line of the light illuminates the sample region in the same position as the resulting emission light through a diorchic mirror (DM) and the imaging spectroscope, and finally reaches the CCD camera (CM).
Fig 2.
After image reconstruction, an integral method was used to accumulate the total intensity in each image of the data file. Binary values are used to replace the pixel values of the region of interest with one and replace the pixel values of background with zero. Finding edges based on binary images generates a mask image.
Fig 3.
Hyperspectral fluorescence images and extraction spectrum.
(a) Precolor overlay image of Anabaena flosahuas. (b) Precolor image of Microcytic aeruginosa (c) Color overlay image of Chlorella sp. (d) Mean spectrum of Anabaena flosahuas (e) Mean spectrum of Microcytic aeruginosa. (f) Mean spectrum of Chlorella sp.
Fig 4.
MCR analysis of algal cells for Anabaena flosahuas.
(a) Concentration image of PBS resulting from MCR analysis of the entire spectral area. (b) Concentration distribution images of Chl-a resulting from analysis of the entire spectral area using MCR. (c) Pure spectral profile of chlorophyll (dotted line) and PBS (solid line).
Fig 5.
MCR analysis of Microcystis aeruginosa.
(a) Concentration image of PBS resulting from analysis of the entire spectral area using MCR. (b) Concentration image of CHl-a resulting from analysis of the entire spectral area using MCR. (c) MCR pure-component spectrum containing the PBS component (dotted line) and chlorophyll-a (solid line). Color scales are identical for all images to facilitate comparison between images. The x-axis and y-axis of the image represent image size (pixels).
Fig 6.
MCR analysis of Chlorella sp. for the entire spectrum.
(a) Concentration image of carotenoids resulting from analysis of the entire spectral area using MCR. (b) Concentration image of CHl-a resulting from analysis of the entire spectral area using MCR. (c) MCR pure-component spectrum containing the PBS component (solid line) and chlorophyll-a (dotted line). Color scales are identical for all images to facilitate comparison between images. The x-axis and y-axis of the image represent image size (pixels).
Fig 7.
MCR analysis of Chlorella sp. for spectrum selection.
(a) Concentration images for Chlorella sp. of carotenoid resulting from analysis of selective spectral area (470–574 nm) (b) Concentration images of Chlorophyll a (c) Concentration images of Light-Harvesting Complex II(LCH-II). (f) pure component spectrum of Chlorella sp. representing chlorophyll a (solid line) and light-harvesting complex II (dotted line) (e) Emission measurement under pigment extraction above 574 nm excited at 512 nm (ex: 512 nm). This describes the intensity difference between carotenoid and chlorophyll spectral areas.
Fig 8.
(a) Emission spectrum and (b) absorbance spectrum of Chlorella sp. Emission measurement under pigment extraction excited at 512 nm (ex: 512 nm). The peak is at 671 nm, 587 nm, 492 nm and 438 nm. This describes the intensity difference between carotenoid and chlorophyll spectral areas.