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Multicolor fluorescence microscopy using static light sheets and a single-channel detection.


ABSTRACT: We present a multicolor fluorescence microscope system, under a selective plane illumination microscopy (SPIM) configuration, using three continuous wave-lasers and a single-channel-detection camera. The laser intensities are modulated with three time-delayed pulse trains that operate synchronously at one third of the camera frame rate, allowing a sequential excitation and an image acquisition of up to three different biomarkers. The feasibility of this imaging acquisition mode is demonstrated by acquiring single-plane multicolor images of living hyphae of Neurospora crassa. This allows visualizing simultaneously the localization and dynamics of different cellular components involved in apical growth in living hyphae. The configuration presented represents a noncommercial, cost-effective alternative microscopy system for the rapid and simultaneous acquisition of multifluorescent images and can be potentially useful for three-dimensional imaging of large biological samples.

SUBMITTER: Licea-Rodriguez J 

PROVIDER: S-EPMC6985699 | biostudies-literature | 2019 Jan

REPOSITORIES: biostudies-literature

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Multicolor fluorescence microscopy using static light sheets and a single-channel detection.

Licea-Rodriguez Jacob J   Figueroa-Melendez Alfredo A   Falaggis Konstantinos K   Plata-Sanchez Marcos M   Riquelme Meritxell M   Rocha-Mendoza Israel I  

Journal of biomedical optics 20190101 1


We present a multicolor fluorescence microscope system, under a selective plane illumination microscopy (SPIM) configuration, using three continuous wave-lasers and a single-channel-detection camera. The laser intensities are modulated with three time-delayed pulse trains that operate synchronously at one third of the camera frame rate, allowing a sequential excitation and an image acquisition of up to three different biomarkers. The feasibility of this imaging acquisition mode is demonstrated b  ...[more]

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