title: Classification of pollen species using autofluorescence image analysis.
abstract: A new method to classify pollen species was developed by monitoring autofluorescence images of pollen grains. The pollens of nine species were selected, and their autofluorescence images were captured by a microscope equipped with a digital camera. The pollen size and the ratio of the blue to red pollen autofluorescence spectra (the B/R ratio) were calculated by image processing. The B/R ratios and pollen size varied among the species. Furthermore, the scatter-plot of pollen size versus the B/R ratio showed that pollen could be classified to the species level using both parameters. The pollen size and B/R ratio were confirmed by means of particle flow image analysis and the fluorescence spectra, respectively. These results suggest that a flow system capable of measuring both scattered light and the autofluorescence of particles could classify and count pollen grains in real time.
authors.affiliation: Electronics and Optics Research Laboratories, Kowa Company, Ltd., 3-3-1 Chofugaoka, Chofu, Tokyo 182-0021, Japan.
authors.forename: Kotaro
authors.initials: K
authors.lastname: Mitsumoto
authors.forename: Katsumi
authors.initials: K
authors.lastname: Yabusaki
authors.forename: Hideki
authors.initials: H
authors.lastname: Aoyagi
mesh_terms: D005434:Flow Cytometry / Q000379:methods; D005453:Fluorescence; D005456:Fluorescent Dyes / Q000494:pharmacology; D007091:Image Processing, Computer-Assisted / Q000379:methods; D008856:Microscopy, Fluorescence / Q000379:methods*; D015233:Models, Statistical; D010316:Particle Size; D011058:Pollen / Q000737:chemistry / Q000145:classification* / Q000235:genetics*; D012542:Scattering, Radiation; D013050:Spectrometry, Fluorescence / Q000379:methods*
chemical_list: D005456:Fluorescent Dyes