Publication Date:
2012-04-25
Description:
Insights into the three-dimensional (3D) organization and function of intracellular structures at nanometer resolution, holds the key to our understanding of the molecular underpinnings of cellular structure−function. Besides this fundamental understanding of the cell at the molecular level, such insights hold great promise in identifying the disease processes by their altered molecular profiles, and help determine precise therapeutic treatments. To achieve this objective, previous studies have employed electron microscopy (EM) tomography with reasonable success. However, a major hurdle in the use of EM tomography is the tedious procedures involved in fixing, high-pressure freezing, staining, serial sectioning, imaging, and finally compiling the EM images to obtain a 3D profile of sub-cellular structures. In contrast, the resolution limit of EM tomography is several nanometers, as compared to just a single or even sub-nanometer using the atomic force microscope (AFM). Although AFM has been hugely successful in 3D imaging studies at nanometer resolution and in real time involving isolated live cellular and isolated organelles, it has had limited success in similar studies involving 3D imaging at nm resolution of intracellular structure–function in situ. In the current study, using both AFM and EM on aldehyde-fixed and semi-dry mouse pancreatic acinar cells, new insights on a number of intracellular structure–function relationships and interactions were achieved. Golgi complexes, some exhibiting vesicles in the process of budding were observed, and small vesicles were caught in the act of fusing with larger vesicles, possibly representing either secretory vesicle biogenesis or vesicle refilling following discharge, or both. These results demonstrate the power and scope of the combined engagement of EM and AFM imaging of fixed semi-dry cells, capable of providing a wealth of new information on cellular structure–function and interactions. Content Type Journal Article Category Original Paper Pages 1-16 DOI 10.1007/s00418-012-0948-x Authors Sunxi Wang, Department of Chemical Engineering and Materials Science, College of Engineering, Wayne State University, 5050 Anthony Wayne Drive, Detroit, MI 48202, USA Jin-Sook Lee, Department of Physiology, School of Medicine, Wayne State University, 540 E. Canfield, Detroit, MI 48201, USA Nicole Bishop, Department of Pathology, Microscopy Imaging Center, University of Vermont College of Medicine, Burlington, VT 05405, USA Aleksandar Jeremic, Department of Physiology, School of Medicine, Wayne State University, 540 E. Canfield, Detroit, MI 48201, USA Won Jin Cho, Department of Physiology, School of Medicine, Wayne State University, 540 E. Canfield, Detroit, MI 48201, USA Xuequn Chen, Department of Physiology, School of Medicine, Wayne State University, 540 E. Canfield, Detroit, MI 48201, USA Guangzhao Mao, Department of Chemical Engineering and Materials Science, College of Engineering, Wayne State University, 5050 Anthony Wayne Drive, Detroit, MI 48202, USA Douglas J. Taatjes, Department of Pathology, Microscopy Imaging Center, University of Vermont College of Medicine, Burlington, VT 05405, USA Bhanu P. Jena, Department of Chemical Engineering and Materials Science, College of Engineering, Wayne State University, 5050 Anthony Wayne Drive, Detroit, MI 48202, USA Journal Histochemistry and Cell Biology Online ISSN 1432-119X Print ISSN 0948-6143
Print ISSN:
0948-6143
Electronic ISSN:
1432-119X
Topics:
Biology
,
Medicine
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