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Individual bioaerosol particle discrimination by multi-photon excited fluorescence

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Abstract

Femtosecond laser induced multi-photon excited fluorescence (MPEF) from individual airborne particles is tested for the first time for discriminating bioaerosols. The fluorescence spectra, analysed in 32 channels, exhibit a composite character originating from simultaneous two-photon and three-photon excitation at 790 nm. Simulants of bacteria aggregates (clusters of dyed polystyrene microspheres) and different pollen particles (Ragweed, Pecan, Mulberry) are clearly discriminated by their MPEF spectra. This demonstration experiment opens the way to more sophisticated spectroscopic schemes like pump-probe and coherent control.

© 2011 Optical Society of America

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Figures (3)

Fig. 1
Fig. 1 Experimental set-up; I1, I2 - air inlets; Y1, Y2 - crossed diode lasers; D1, D2 - spectrally filtered photomultipliers; SSA - scatter signal analyzer; CFG - constant frequency generator; LO - Ti:Sa femtosecond oscillator; LC - regenerative amplifier; PL -pump Nd:YLF laser; PC1, PC2 - injection and extraction Pockels cells; SDG - system delay generator; L1 - reflective objective; G1 - grating; D3 - 32-anodes PMT; SAS - spectrum acquisition electronics.
Fig. 2
Fig. 2 MPEF microscopy images of a) FB345 clusters, b) Ragweed pollens, c) Pecan pollens, and d) Mulberry pollens. The large images correspond to the UV fluorescence channel (360 nm, FWHM 12 nm); the insets display the composite images of the UV- (360 nm, FWHM 12 nm), Blue- (485 nm, FWHM 20 nm), Green- (531 nm, FWHM 40 nm) and Red-channels (600 nm, FWHM 70 nm). The reference ruler corresponds to 50 μm.
Fig. 3
Fig. 3 Single-shot MPEF spectra of individual aerosol particles. (a) Simulants of bacteria aggregates (FB345); (b) Ragweed pollen; (c) Pecan Pollen; and (d) Mulberry pollen. The dashed lines represent the 96% confidence interval (±2σ) calculated from a series of individual detection events.

Tables (1)

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Table 1 Detection statistics

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