+ Site Statistics
+ Search Articles
+ PDF Full Text Service
How our service works
Request PDF Full Text
+ Follow Us
Follow on Facebook
Follow on Twitter
Follow on LinkedIn
+ Subscribe to Site Feeds
Most Shared
PDF Full Text
+ Translate
+ Recently Requested

High resolution ultrastructural mapping of total calcium: Electron spectroscopic imaging/electron energy loss spectroscopy analysis of a physically/chemically processed nerve-muscle preparation



High resolution ultrastructural mapping of total calcium: Electron spectroscopic imaging/electron energy loss spectroscopy analysis of a physically/chemically processed nerve-muscle preparation



Proceedings of the National Academy of Sciences of the United States of America 93(10): 4799-4803



We report on a procedure for tissue preparation that combines thoroughly controlled physical and chemical treatments: quick-freezing and freeze-drying followed by fixation with OSO-4 vapors and embedding by direct resin infiltration. Specimens of frog cutaneous pectoris muscle thus prepared were analyzed for total calcium using electron spectroscopic imaging/electron energy loss spectroscopy (ESI/EELS) approach. The preservation of the ultrastructure was excellent, with positive K/Na ratios revealed in the fibers by x-ray microanalysis. Clear, high-resolution EELS/ESI calcium signals were recorded from the lumen of terminal cisternae of the sarcoplasmic reticulum but not from longitudinal cisternae, as expected from previous studies carried out with different techniques. In many mitochondria, calcium was below detection whereas in others it was appreciable although at variable level. Within the motor nerve terminals, synaptic vesicles as well as some cisternae of the smooth endoplasmic reticulum yielded positive signals at variance with mitochondria, that were most often below detection. Taken as a whole, the present study reveals the potential of our experimental approach to map with high spatial resolution the total calcium within individual intracellular organelles identified by their established ultrastructure, but only where the element is present at high levels.

Please choose payment method:






(PDF emailed within 0-6 h: $19.90)

Accession: 008772900

Download citation: RISBibTeXText

PMID: 8643483

DOI: 10.2307/38856


Related references

High resolution mapping of total calcium in osteoblasts by electron energy loss spectroscopy and electron spectroscopic imaging. Biology of the Cell 91(3): 276-276, 1999

High resolution micro analysis of biological specimens by electron energy loss spectroscopy and electron spectroscopic imaging. Journal of Electron Microscopy 31(1): 104-105, 1982

High resolution micro analysis of biological specimens by electron energy loss spectroscopy and by electron spectroscopic imaging. Journal of Ultrastructure Research 72(3): 336-348, 1980

Ultrastructural detection of calcium and magnesium in the chromatoid body of mouse spermatids by electron spectroscopic imaging and electron energy loss spectroscopy. Journal of Histochemistry and Cytochemistry 41(8): 1155-1162, 1993

Calcium distribution in high-pressure frozen bone cells by electron energy loss spectroscopy and electron spectroscopic imaging. Histochemistry & Cell Biology. 109(2): 167-174,., 1998

Quantitative compositional imaging of cardiac muscle at 5-10 nm resolution with electron energy loss spectroscopic - and X-ray-mapping. Molecular Biology of the Cell 13(Supplement): 137a-138a, 2002

Ultrastructural aluminium detection in amphibian tissues by electron spectroscopic imaging and electron energy-loss spectroscopy. Journal of Microscopy 162(Pt 1): 115-122, 1991

Ultrastructural distribution of terbium across capillary endothelium: detection by electron spectroscopic imaging and electron energy loss spectroscopy. Journal of Histochemistry and Cytochemistry 38(2): 275-282, 1990

New applications of electron spectroscopic imaging esi electron energy loss spectroscopy eels and electron spectroscopic diffraction esd including computer assisted image processing. European Journal of Cell Biology Supplement (18): 14, 1987

Demonstration of calcium in dermal melanocytes of Xenopus laevis and Poecilia reticulata with electron energy-loss spectroscopy and electron spectroscopic imaging. Journal of Microscopy 162(Pt 1): 141-146, 1991

Localization of calcium in the cyanobiont and gonidial zone of Cycas revoluta Thunb. by microelectrodes, chlorotetracycline, electron spectroscopic imaging and electron energy loss spectroscopy. Protoplasma 179(3/4): 151-157, 1994

Quantification of total calcium in terminal cisternae of skinned muscle fibers by imaging electron energy-loss spectroscopy. Journal of Muscle Research & Cell Motility 20(5-6): 505-515, 1999

Em 902 a new transmission electron microscopy for microanalysis with electron spectroscopic imaging and electron energy loss spectroscopy. Ultramicroscopy: 386, 1984

Cytochemical analysis and localization of a H pylori P type ATPase expressed in E coli by electron spectroscopic imaging and electron energy loss spectroscopy. Gastroenterology 110(4 SUPPL ): A960, 1996

High-resolution calcium mapping of the endoplasmic reticulum-Golgi-exocytic membrane system. Electron energy loss imaging analysis of quick frozen-freeze dried PC12 cells. Molecular Biology of the Cell 8(8): 1501-1512, 1997