ArticleBiomedical optics express2016
Simultaneous multicolor imaging of wide-field epi-fluorescence microscopy with four-bucket detection.
Article in Biomedical optics express, 2016. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 4 papers.
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Who cites it
4 citing papers in PubMed.
- Microscopy is better in color: development of a streamlined spectral light path for real-time multiplex fluorescence microscopy.Biomedical optics express · 2022Article
- Serial imaging of micro-agents and cancer cell spheroids in a microfluidic channel using multicolor fluorescence microscopy.PloS one · 2021Article
- Enhancing the Value of Histopathological Assessment of Allograft Biopsy Monitoring.Transplantation · 2019Review
- Investigating dye performance and crosstalk in fluorescence enabled bioimaging using a model system.PloS one · 2017Article
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Authors and funding
5 authors.
Funding
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Abstract
We demonstrate simultaneous imaging of multiple fluorophores using wide-field epi-fluorescence microscopy with a monochrome camera. The intensities of the three lasers are modulated by a sinusoidal waveform in order to excite each fluorophore with the same modulation frequency and a different time-delay. Then, the modulated fluorescence emissions are simultaneously detected by a camera operating at four times the excitation frequency. We show that two different fluorescence beads having crosstalk can be clearly separated using digital processing based on the phase information. In addition, multiple organelles within multi-stained single cells are shown with the phase mapping method, demonstrating an improved dynamic range and contrast compared to the conventional fluorescence image. These findings suggest that wide-field epi-fluorescence microscopy with four-bucket detection could be utilized for high-contrast multicolor imaging applications such as drug delivery and fluorescence in situ hybridization.
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