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

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...

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Bibliographic Details
Published in:Journal of biomedical optics 2019-01, Vol.24 (1), p.1-8
Main Authors: Licea-Rodriguez, Jacob, Figueroa-Melendez, Alfredo, Falaggis, Konstantinos, Plata-Sanchez, Marcos, Riquelme, Meritxell, Rocha-Mendoza, Israel
Format: Article
Language:English
Subjects:
Biological properties
Biological samples
Biomarkers
Biomarkers - metabolism
Cameras
Color
Configurations
Continuous radiation
Equipment Design
Fluorescence
Fluorescence microscopy
Fluorescent Dyes - chemistry
Green Fluorescent Proteins - chemistry
Hyphae
Image acquisition
Image Processing, Computer-Assisted - methods
Imaging, Three-Dimensional - methods
Lasers
Light
Light sheets
Localization
Luminescent Proteins - chemistry
Microscopy
Microscopy, Fluorescence - instrumentation
Microscopy, Fluorescence - methods
Neurospora
Neurospora crassa - metabolism
Red Fluorescent Protein
Rhodamines - chemistry
Three dimensional imaging
Transistors
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Description
Summary: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.
ISSN:1083-3668
1560-2281
DOI:10.1117/1.JBO.24.1.016501