Development of a novel real-time pollen-sorting counter using species-specific pollen autofluorescence

Abstract

We developed a novel flow particle analyzer that automatically classifies airborne pollen grains. The design of the particle counter (model KP-1000) is based on that of a flow cytometer, applied to the measurement of airborne particles. The counter classifies pollen species by simultaneously detecting both scattered light and the characteristic fluorescence excited by ultraviolet light in the flow cell. We observed airborne pollen using KP-1000 pollen counters and Durham samplers to compare their performance at three study sites in Japan during the spring pollen season. The pollen counter followed the variation in pollen concentrations, and its daily pollen counts were significantly correlated with the results of the Durham sampling method at all study sites. Although the counter over-or under-counted 2 target pollen species (Cryptomeria japonica and Chamaecyparis obtusa) when they coexisted, a data correction based on the Durham sampling results improved the accuracy of pollen classification of the counter. Our results indicate that the new pollen counter has a strong potential for counting and identifying airborne pollen grains in real time, and it requires further improvement, field trials, and tests with other common airborne pollen grains.

Detection and characterization of biological and other organic-carbon aerosol particles in atmosphere using fluorescence

“…In general, LIF based systems promise to be able to differentiate bioaerosol particles from most non-bioaerosol particles (e.g., [27,29,19,83,62,65,41]); discriminate biothreat agents from atmospheric aerosol particles; and classify different bioaerosol species such as pollen versus fungi, as shown in Fig. 3(a) [42,58,71,97,44,45,28]. But keep in mind, not all fluorescent particles are bioaerosols, and also some biomolecules contained aerosol particles may be below the fluorescence detection threshold.…”

Fullerene fine particles adhere to pollen grains and affect their autofluorescence and germination

“…17,18 Fluorescence and bright-field micrographs of pollen grains on a glass plate were taken using a fluorescence microscope (Leica DMR; Leica Microsystems, Tokyo, Japan) equipped with both an ultraviolet illuminating system using a xenon lamp and a band-pass filter (central wavelength: 340 nm) and CCD camera (SPOT ISA-CE Color; Diagnostic Instruments, Inc, Sterling Heights, MI). Samples for scanning electron microscopy study were prepared according to the method of Vinckier and Smets.…”

Photophoretic trapping-Raman spectroscopy for single pollens and fungal spores trapped in air

“…Some of these instruments measure single particle elastic scattering, and/or single particle fluorescence, and/or laser-or spark-induced breakdown spectroscopy (LIBS/SIBS) and some are commercially available [3]. Laser induced fluorescence (LIF), especially dual-wavelength UV-LIF has been demonstrated for near-real time detection and partial classification of bioaerosols particles [4][5][6][7][8][9]. The technique was shown to be capable of differentiating pollens from various plant species [9,10].…”

“…Laser induced fluorescence (LIF), especially dual-wavelength UV-LIF has been demonstrated for near-real time detection and partial classification of bioaerosols particles [4][5][6][7][8][9]. The technique was shown to be capable of differentiating pollens from various plant species [9,10]. Fluorescence clustering analysis, especially when it is combined with the dualwavelength UV-LIF, appears promising for rapid airborne bioaerosol characterization [11,12].…”