The longitudinal electric field produced by focusing a radially polarized beam is applied in confocal laser scanning microscopy by introducing a higher-order transverse mode, combined with a technique of polarization conversion for signal detection. This technique improves signal detection corresponding to the longitudinally polarized field under a small confocal pinhole, enabling full utilization of the small focal spot characteristic of the longitudinal field. Detailed numerical and experimental studies demonstrate the enhanced spatial resolution in confocal imaging that detects a scattering signal using a higher-order radially polarized beam. Our method can be widely applied in various imaging techniques that detect coherent signals such as second-harmonic generation microscopy.

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BACKGROUND: High-levels of etoxazole resistance have not yet been frequently reported in Panonychus citri. Although a highly resistant strain was discovered in 2014, etoxazole resistance has not become a significant problem in areas of citrus production in Japan. A target site mutation in chitin synthase 1 (CHS1), I1017F, is a major etoxazole-resistance factor in Tetranychus urticae.To investigate the mechanisms of etoxazole resistance and the dispersal of resistance genes, we analyzed target-site mutations in a highly resistant strain and their geographical distribution in Japan.RESULTS: High-level etoxazole resistance was completely recessive. The I1017F mutation was detected in CHS1 of the highly resistant strain, and its frequency was correlated with the hatchability of eggs treated with etoxazole. Sequencing and variant frequency analyses of local populations by quantitative polymerase chain reaction revealed that I1017F is restricted to the Ariake Sea area of Kyushu Island. Although a new nonsynonymous substitution, S1016L, accompanied by I1017F was found in CHS1 of the highly resistant strain, CRISPR/Cas9 engineering of flies showed that S1016L had no effect on the etoxazole resistance conferred by I1017F.CONCLUSION: I1017F is a major target site mutation that confers high-level etoxazole resistance on P. citri. Dispersion of I1017F possibly was suppressed as a result of the completely recessive inheritance of resistance together with low gene flow between local populations.

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