Practical Solutions to Frequent Problems Encountered in Thin Sections Electron Microscopy

Lavorato, Manuela; Franzini‐Armstrong, Clara

doi:10.1017/s1551929517000438

Cited by 7 publications

(7 citation statements)

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“…034-LCS-2019). 20 The tissue was dehydrated in ethanol and acetone and embedded in Epon. The heart was removed quickly and placed in solution containing the following (in mM): 130 NaCl, 24 NaHCO 3 , 1.2 NaH 2 PO 4 , 1.0 MgCl 2 , 1.8 CaCl 2 , 4.0 KCl, and 5.6 glucose equilibrated with 95% O 2 -5% CO 2 (pH 7.4 at 35.5°C).…”

Section: Methodsmentioning

confidence: 99%

“…The tissue was rinsed in cacodylate buffer, and either postfixed in 2% OsO 4 in the same buffer containing 0.6% K 3 Fe(CN) 6 , or postfixed in 2% OsO4 in the same buffer for 1 hour at room temperature, rinsed in H 2 O and en-bloc stained in aqueous saturated uranyl acetate for 1 hour. 20 The tissue was dehydrated in ethanol and acetone and embedded in Epon. Thin (50-60 nm) sections were cut at right angles to the thin layer of the SA node on a Leica Sitte microtome and stained with "Sato" lead citrate.…”

Section: Methodsmentioning

confidence: 99%

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Ultrastructure of primary pacemaking cells in rabbit sino‐atrial node cells indicates limited sarcoplasmic reticulum content

et al. 2020

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The main mammalian heart pacemakers are spindle-shaped cells compressed into tangles within protective layers of collagen in the sino-atrial node (SAN). Two cell types, "dark" and "light," differ on their high or low content of intermediate filaments, but share scarcity of myofibrils and a high content of glycogen. Sarcoplasmic reticulum (SR) is scarce. The free SR (fSR) occupies 0.04% of the cell volume within ~0.4 µm wide peripheral band. The junctional SR (jSR), constituting peripheral couplings (PCs), occupies 0.03% of the cell volume. Total fSR + jSR volume is 0.07% of cell volume, lower than the SR content of ventricular myocytes. The average distance between PCs is 7.6 µm along the periphery. On the average, 30% of the SAN cells surfaces is in close proximity to others. Identifiable gap junctions are extremely rare, but small sites of close membrane-to-membrane contacts are observed. Possibly communication occurs via these very small sites of contact if conducting channels (connexons) are located within them. There is no obvious anatomical detail that might support ephaptic coupling. These observations have implications for understanding of SAN cell physiology, and require incorporation into biophysically detailed models of SAN cell behavior that currently do not include such features. K E Y W O R D S gap junctions, pacemaker cells, sarcoplasmic reticulum, sino-atrial node c Bossen et al, 1978, 33 d Mobley and Eisenberg, 1975 34 e (Number of hearts/Number of cells/Number of measurements)

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Ultrastructure of primary pacemaking cells in rabbit sino‐atrial node cells indicates limited sarcoplasmic reticulum content

et al. 2020

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“…Thus the above descriptions are not derived from a systematic study. Lavorato et al, 2016;Castellani et al, 1989;Lavorato and Franzini-Armstrong 2017.…”

Section: Glycogenmentioning

confidence: 99%

“…See also 2.8.6. Franzini- Armstrong 2007;Huang et al, 2013;Lavorato et al, 2017Lavorato et al, , 2020.…”

Section: Kissing Junctions and Nano Tunnelsunclassified

“…A RyR2 mutation linked to catecholaminergic polymorphic ventricular tachycardia (A4860G) induces a calcium imbalance by depressing RyR2 channel activity during excitation-contraction coupling. Cardiac myocytes of heterozygous mice carrying the mutation exhibit an unusual structural response: an increase in the frequency of mitochondrial nanotunnels (4.3.2 A) that allow a slow exchange between mitochondria, see 2.8.5 D. Lavorato et al, 2017.…”

Section: Mitochondria-related Mutationmentioning

confidence: 99%

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Muscle: 50+ Years of Electron Microscopy

Franzini‐Armstrong

2021

Preprint

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This project was initiated with the aim of preserving for the future a portion of the extensive muscle ultrastructural image archive accumulated over a 50 year period by the author and her laboratory. Between the late 1950s and current times electron microscopy (EM) evolved. The early EM revealed the ultrastructural identification and definition of cell organelles and laid the foundation for cell biology. This continues with he discovery of new structural organizations and organelles. Currently EM images provide the essential basis for identification of structural changes associated with mutational experiments and newly discovered pathology. The most recent development of techniques allowing near atomic level of resolution brings microscopy to modern times. This evolution has been greatly enriched by the rebirth of light microscopy, fueled by dynamic views of events based on video techniques, by the availability of specific targeting of cell components and by confocal microscopy that allows 3-D structural reconstructs. Correlation between light and electron microscopy, when appropriately done, is highly revealing.The images in this publication present descriptive and structure-function correlations covering the whole animal kingdom with some areas more extensively illustrated than others. The first part classifies the images based in their site of origin, the second part identifies functional correlations, the third part introduces results of mutational experiments, the fourth part covers aging and pathology examples, the fifth part is a guide to the techniques. The images are presented with a numerical identification correlated with a very succinct description. References are limited to the directly relevant published material from the author's laboratory that are helpful in extending the limited legends and in assigning contributions. With very few exceptions, references to the literature are not given. The author asks for the forgiveness of the many friends, trainees, collaborators and competitors whose contributions and priorities are not recognized.The work has been stimulated by interactions with teachers and advisors. Keith R. Porter, father of the endoplasmic reticulum and sarcoplasmic reticulum instilled admiration for Nature's work and was always been a guiding light in the author's work. Richard J. Podolsky and Andrew F. Huxley taught the importance of structural correlations. Past and present trainees and collaborators, whose names appear in the reference list, have added depth of understanding.In the case of skeletal muscle much necessary information is gained by using the muscle bible: "Myology", A.G. Engel and C. Franzini-Armstrong, Eds. 3rd Edition, McGraw Hill, N.Y. 2004.

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Artifacts and Pitfalls in Electron Microscopy of the Kidney

Haro

2023

Atlas of Renal Ultrastructural Pathology

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Practical Solutions to Frequent Problems Encountered in Thin Sections Electron Microscopy

Cited by 7 publications

References 4 publications

Ultrastructure of primary pacemaking cells in rabbit sino‐atrial node cells indicates limited sarcoplasmic reticulum content

Ultrastructure of primary pacemaking cells in rabbit sino‐atrial node cells indicates limited sarcoplasmic reticulum content

Muscle: 50+ Years of Electron Microscopy

Artifacts and Pitfalls in Electron Microscopy of the Kidney

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