Dynamic measurement of fluorescent proteins spectral distribution on virus infected cells

Ja-Yun Lee; Ming-Xiu Wu; Chia-Yun Kao; Tzong-Yuan Wu; I-Jen Hsu

doi:10.1117/12.707490

8 September 2006 Dynamic measurement of fluorescent proteins spectral distribution on virus infected cells

Ja-Yun Lee, Ming-Xiu Wu, Chia-Yun Kao, Tzong-Yuan Wu, I-Jen Hsu

Author Affiliations +

Proceedings Volume 6343, Photonics North 2006; 63430E (2006) https://doi.org/10.1117/12.707490
Event: Photonics North 2006, 2006, Quebec City, Canada

Abstract

We constructed a dynamic spectroscopy system that can simultaneously measure the intensity and spectral distributions of samples with multi-fluorophores in a single scan. The system was used to monitor the fluorescence distribution of cells infected by the virus, which is constructed by a recombinant baculoviruses, vAcD-Rhir-E, containing the red and green fluorescent protein gene that can simultaneously produce dual fluorescence in recombinant virus-infected Spodoptera frugiperda 21 cells (Sf21) under the control of a polyhedrin promoter. The system was composed of an excitation light source, a scanning system and a spectrometer. We also developed an algorithm and fitting process to analyze the pattern of fluorescence distribution of the dual fluorescence produced in the recombinant virus-infected cells. All the algorithm and calculation are automatically processed in a visualized scanning program and can monitor the specific region of sample by calculating its intensity distribution. The spectral measurement of each pixel was performed at millisecond range and the two dimensional distribution of full spectrum was recorded within several seconds. We have constructed a dynamic spectroscopy system to monitor the process of virus-infection of cells. The distributions of the dual fluorescence were simultaneously measured at micrometer resolution.

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Ja-Yun Lee, Ming-Xiu Wu, Chia-Yun Kao, Tzong-Yuan Wu, and I-Jen Hsu "Dynamic measurement of fluorescent proteins spectral distribution on virus infected cells", Proc. SPIE 6343, Photonics North 2006, 63430E (8 September 2006); https://doi.org/10.1117/12.707490

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KEYWORDS

Luminescence

Spectroscopy

Algorithm development

Fluorescent proteins

Image processing

3D image processing

Dynamical systems

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