Monitoring and quantification of the protein partition during cytokinesis with fluorescent spectral imaging

Ja-Yun Lee; Yi-Ting Lin; Tzong-Yuan Wu; I-Jen Hsu

doi:10.1117/12.808802

16 February 2009 Monitoring and quantification of the protein partition during cytokinesis with fluorescent spectral imaging

Ja-Yun Lee, Yi-Ting Lin, Tzong-Yuan Wu, I-Jen Hsu

Author Affiliations +

Proceedings Volume 7191, Fluorescence In Vivo Imaging Based on Genetically Engineered Probes: From Living Cells to Whole Body Imaging IV; 71910E (2009) https://doi.org/10.1117/12.808802
Event: SPIE BiOS, 2009, San Jose, California, United States

Abstract

Cytokinesis is a consecutive process during cell division. For systems biological studies, it is important to precisely monitor and quantify proteins in different cell stages and mitosis processes. However, the absolute quantities in living cells are usually difficult to quantify. Fluorescent protein tagged protein is one of the techniques that are usually applied to monitor biological behaviors and phenomena. In this study, an insect cell line, DPnE, which can stably express both green fluorescent protein (EGFP) and red fluorescent protein (DsRed) was established. This dual fluorescent cell line was chosen as a model system to monitor the protein partition during cytokinesis. A spectrum analysis system was established and integrated in an inverted microscope. The two-dimensional distribution of the full fluorescent spectra of the two fluorescent proteins was obtained in a time-lapse series. Furthermore, we also developed an algorithm to analyze the quantities of both fluorescent proteins in the daughter cells and parent cells during the process of cytokinesis, respectively. With this innovative optical system and algorithm, the proteins partition during cytokinesis can be monitored and quantified precisely.

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Ja-Yun Lee, Yi-Ting Lin, Tzong-Yuan Wu, and I-Jen Hsu "Monitoring and quantification of the protein partition during cytokinesis with fluorescent spectral imaging", Proc. SPIE 7191, Fluorescence In Vivo Imaging Based on Genetically Engineered Probes: From Living Cells to Whole Body Imaging IV, 71910E (16 February 2009); https://doi.org/10.1117/12.808802

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KEYWORDS

Proteins

Fluorescent proteins

Green fluorescent protein

Algorithm development

Microscopes

Imaging spectroscopy

Imaging systems

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