Studies of cell division (mitosis and cytokinesis) by dynamic secondary ion mass spectrometry ion microscopy: LLC-PK1 epithelial cells as a model for subcellular isotopic imaging.
作者： S. Chandra
DOI: 10.1046/J.1365-2818.2001.00944.X
关键词: Cell division 、 Nocodazole 、 Mitosis 、 Chemistry 、 Cytokinesis 、 Fluorescence-lifetime imaging microscopy 、 Secondary ion mass spectrometry 、 Cell biology 、 Cell culture 、 Organelle
摘要: The feasibility of the renal epithelial LLC-PK1 cell line as a model for division studies with secondary ion mass spectrometry (SIMS) was tested. In this line, cells undergoing all stages mitosis and cytokinesis remained firmly attached to substrate could be cryogenically prepared. Fractured freeze-dried mitotic showed well-preserved organelles revealed by fluorescence imaging rhodamine-123 C6-NBD-ceramide confocal laser scanning microscopy. Secondary electron microscopy analysis fractured dividing minimal surface topography that does not interfere in isotopic both positive (39K, 23Na, 24Mg, 40Ca, etc.) negative (31P, 35Cl, secondaries CAMECA IMS-3f microscope. Mitotic intracellular ionic composition even most diffusible ions (total concentrations 39K+ 23Na+) K : Na ratios approximately 10. Structurally damaged identified their reduced an excessive loading calcium. Quantitative three-dimensional SIMS required studying subcellular calcium distribution cells. also allowed M-phase arrested mitosis-arresting drugs (taxol, monastrol nocodazole). This study opens new avenues research related fluxes chemical SIMS.
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