Effective Cellular Morphology Analysis for Differentiation Processes by a Fluorescent 1,3a,6a-Triazapentalene Derivative Probe in Live Cells

• Published in: PloS one Vol. 11; no. 8; p. e0160625
• Main Authors: Kamada, Rui, Tano, Fumi, Kudoh, Fuki, Kimura, Nozomi, Chuman, Yoshiro, Osawa, Ayumi, Namba, Kosuke, Tanino, Keiji, Sakaguchi, Kazuyasu
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 04.08.2016; Public Library of Science (PLoS)
• Subjects: 3T3-L1 Cells; A549 Cells; Animals; Biological activity; Biological effects; Biology and Life Sciences; Bridged Bicyclo Compounds, Heterocyclic - chemistry; Cancer; Cell culture; Cell Cycle; Cell Differentiation; Cell Line, Tumor; Cell morphology; Cell Nucleus - chemistry; Cell Nucleus - metabolism; Cytology; Cytoplasm; Cytoplasm - chemistry; Cytoplasm - metabolism; Dendritic spines; Dendritic structure; Differentiation (biology); Fluorescence; Fluorescent Dyes - chemistry; Genetic aspects; HEK293 Cells; Humans; Information processing; Laboratories; Leukemia; Leukocytes (neutrophilic); Medical diagnosis; Mice; Microscopy; Microscopy, Fluorescence; Morphology; Organic chemistry; PC12 Cells; Penicillin; Phagocytosis; Physical Sciences; Physiological aspects; Rats; Research and Analysis Methods; Science; Signal processing; Social Sciences; Spectrometry, Fluorescence; Spine; Stem cells; Toxicity; Visualization; Japan
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0160625

Nuclear and cytoplasmic morphological changes provide important information about cell differentiation processes, cell functions, and signal responses. There is a strong desire to develop a rapid and simple method for visualizing cytoplasmic and nuclear morphology. Here, we developed a novel and rapid method for probing cellular morphological changes of live cell differentiation process by a fluorescent probe, TAP-4PH, a 1,3a,6a-triazapentalene derivative. TAP-4PH showed high fluorescence in cytoplasmic area, and visualized cytoplasmic and nuclear morphological changes of live cells during differentiation. We demonstrated that TAP-4PH visualized dendritic axon and spine formation in neuronal differentiation, and nuclear structural changes during neutrophilic differentiation. We also showed that the utility of TAP-4PH for visualization of cytoplasmic and nuclear morphologies of various type of live cells. Our visualizing method has no toxicity and no influence on the cellular differentiation and function. The cell morphology can be rapidly observed after addition of TAP-4PH and can continue to be observed in the presence of TAP-4PH in cell culture medium. Moreover, TAP-4PH can be easily removed after observation by washing for subsequent biological assay. Taken together, these results demonstrate that our visualization method is a powerful tool to probe differentiation processes before subsequent biological assay in live cells.