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Electron microscopy of primary cell cultures in solution and correlative optical microscopy using ASEM.
- Source :
-
Ultramicroscopy . Aug2014, Vol. 143, p52-66. 15p. - Publication Year :
- 2014
-
Abstract
- Abstract: Correlative light-electron microscopy of cells in a natural environment of aqueous liquid facilitates high-throughput observation of protein complex formation. ASEM allows the inverted SEM to observe the wet sample from below, while an optical microscope observes it from above quasi-simultaneously. The disposable ASEM dish with a silicon nitride (SiN) film window can be coated variously to realize the primary-culture of substrate-sensitive cells in a few milliliters of culture medium in a stable incubator environment. Neuron differentiation, neural networking, proplatelet-formation and phagocytosis were captured by optical or fluorescence microscopy, and imaged at high resolution by gold-labeled immuno-ASEM with/without metal staining. Fas expression on the cell surface was visualized, correlated to the spatial distribution of F-actin. Axonal partitioning was studied using primary-culture neurons, and presynaptic induction by GluRĪ“2-N-terminus-linked fluorescent magnetic beads was correlated to the presynaptic-marker Bassoon. Further, megakaryocytes secreting proplatelets were captured, and P-selectins with adherence activity were localized to some of the granules present by immuno-ASEM. The phagocytosis of lactic acid bacteria by dendritic cells was also imaged. Based on these studies, ASEM correlative microscopy promises to allow the study of various mesoscopic-scale dynamics in the near future. [Copyright &y& Elsevier]
Details
- Language :
- English
- ISSN :
- 03043991
- Volume :
- 143
- Database :
- Academic Search Index
- Journal :
- Ultramicroscopy
- Publication Type :
- Academic Journal
- Accession number :
- 96188886
- Full Text :
- https://doi.org/10.1016/j.ultramic.2013.10.010