Three-Dimensional Analysis of Tranverse Tubules in Normal and Failing Heart: A Combined Confocal and High Voltage Electron Microscope Study

Early electron microscopic studies documented that significant changes in the membrane systems of cardiac cells occur in both ischemic and non-ischemic heart failure. These studies relied on analysis of two-dimensional sections and although quantitative changes were observed, the overall organization of the tranverse tubules (T-tubules) and the sarcoplasmic reticulum could not be assessed. In a 3-dimensional study using high voltage electron microscopy (EM) of the T-tubules in spontaneously hypertensive rats, Nakamura and Hama (1991) observed that concomitant with an increase in surface area, the T-tubule system becomes progressively more disorganized and exhibits structural irregularities such as increased numbers of longitudinal tubules, numerous short dead end branches and complex tubular aggregates. These authors suggested that this disorganization may interfere with synchronous contraction over the entire cell.In the present study, we examined the 3-dimensional organization of T-tubules in the left ventricle of explanted human hearts using confocal microscopy and EM tomography.

Download Full-text

Three-Dimensional Visualization of the T-System of Frog Muscle Using High Voltage Electron Microscopy and a Lanthanum Stain

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100080286 ◽

1977 ◽

Vol 35 ◽

pp. 570-571 ◽

Cited By ~ 2

Author(s):

Lee D. Peachey ◽

Clara Franzini-Armstrong

Keyword(s):

Electron Microscopy ◽

High Voltage ◽

Three Dimensional ◽

Biological Tissues ◽

High Voltage Electron Microscopy ◽

Transverse Tubular System ◽

Peroxidase Reaction ◽

Voltage Electron ◽

High Voltage Electron ◽

T System

The effective study of biological tissues in thick slices of embedded material by high voltage electron microscopy (HVEM) requires highly selective staining of those structures to be visualized so that they are not hidden or obscured by other structures in the image. A tilt pair of micrographs with subsequent stereoscopic viewing can be an important aid in three-dimensional visualization of these images, once an appropriate stain has been found. The peroxidase reaction has been used for this purpose in visualizing the T-system (transverse tubular system) of frog skeletal muscle by HVEM (1). We have found infiltration with lanthanum hydroxide to be particularly useful for three-dimensional visualization of certain aspects of the structure of the T- system in skeletal muscles of the frog. Specifically, lanthanum more completely fills the lumen of the tubules and is denser than the peroxidase reaction product.

Download Full-text

High-voltage electron microscopy of maxillary gland of spirochete-infected brine shrimp: lesion of efferent tubule

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100103875 ◽

1988 ◽

Vol 46 ◽

pp. 362-363

Author(s):

G. E. Tyson ◽

M. J. Song

Keyword(s):

Electron Microscopy ◽

High Voltage ◽

Brine Shrimp ◽

Natural Populations ◽

Three Dimensional ◽

Dimensional Structure ◽

High Voltage Electron Microscopy ◽

Voltage Electron ◽

High Voltage Electron ◽

Infected Adult

Natural populations of the brine shrimp, Artemia, may possess spirochete- infected animals in low numbers. The ultrastructure of Artemia's spirochete has been described by conventional transmission electron microscopy. In infected shrimp, spirochetal cells were abundant in the blood and also occurred intra- and extracellularly in the three organs examined, i.e. the maxillary gland (segmental excretory organ), the integument, and certain muscles The efferent-tubule region of the maxillary gland possessed a distinctive lesion comprised of a group of spirochetes, together with numerous small vesicles, situated in a cave-like indentation of the base of the tubule epithelium. in some instances the basal lamina at a lesion site was clearly discontinuous. High-voltage electron microscopy has now been used to study lesions of the efferent tubule, with the aim of understanding better their three-dimensional structure.Tissue from one maxillary gland of an infected, adult, female brine shrimp was used for HVEM study.

Download Full-text

High-voltage cytochemistry for three-dimensional observation of nuclei and calculation of total numbers of nuclear pores in retinal Mueller glia

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s042482010016234x ◽

1990 ◽

Vol 48 (3) ◽

pp. 956-957

Author(s):

H. Ishigooka ◽

S. Ueno ◽

L.M. Hjelmeland ◽

M.B. Landers ◽

K. Ogawa

Keyword(s):

Nuclear Envelope ◽

High Voltage ◽

Three Dimensional ◽

Reaction Medium ◽

Nuclear Pores ◽

High Voltage Electron Microscopy ◽

Voltage Electron ◽

High Voltage Electron ◽

G6pase Activity ◽

Mueller Glia

Introduction: We have demonstrated that Glucose-6-phosphatase (G6Pase) activity is localized to the endoplasmic reticulum and nuclear envelope of Mueller glia in the normal and pathological guinea pig retina. Using a combination of this cytochemical technique and high voltage electron microscopy, the distribution of nuclear pores could be clearly observed on the nuclear envelope of Mueller glia because of their anatomical lack of reaction products. This technique was developed to study the three-dimensional structure of nuclei and to calculate total numbers of nuclear pores utilizing a computer graphic analysis system in the normal and pathological retina.Materials and methods: Normal and photocoagulated retina of pigmented adult guinea pigs were perfused with a cold mixture of 0.25% glutaraldehyde and 2% paraformaldehyde in 0.1M cacodylate buffer, and the enucleated globes were hemisected and immersed in the same fixative for 30 min. After sectioning and incubation in the reaction medium for the detection of G6Pase activity by the method of Wachstein-Meisel, the sections were postfixed, dehydrated and embedded in Spurr’s epoxy resin. Serial thick sections (1.0um) were prepared for the observation by a Hitachi high voltage electron microscope (H 1250-M) with an accelerating voltage of 1000 Kv. and pictures were analyzed and three-dimensionally reconstructed by TRI (RATOC Co., Ltd.).

Download Full-text

High-voltage electron microscopic studies of inflammatory cell attachment during blood-brain barrier inflammation

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100158935 ◽

1990 ◽

Vol 48 (3) ◽

pp. 276-277

Author(s):

A.S. Lossinsky ◽

M.J. Song

Keyword(s):

High Voltage ◽

Inflammatory Cell ◽

Cell Attachment ◽

Electron Microscopic ◽

Vascular Perfusion ◽

Tissue Samples ◽

High Voltage Electron Microscopy ◽

Thin Sections ◽

Voltage Electron ◽

High Voltage Electron

Previous studies have suggested the usefulness of high-voltage electron microscopy (HVEM) for investigating blood-bram barrier (BBB) injury and the mechanism of inflammatory-cell (IC) attachment. These studies indicated that, in evaluating standard conventional thin sections, one might miss cellular attachment sites of ICs in their process of attaching to the luminal endothelial cell (EC) surface of cerebral blood vessels. Our current studies in animals subjected to autoimmune disease suggest that HVEM may be useful in localizing precise receptor sites involved in early IC attachment.Experimental autoimmune encephalomyelitis (EAE) was induced in mice and rats according to standard procedures. Tissue samples from cerebellum, thalamus or spinal cords were embedded in plastic following vascular perfusion with buffered aldehyde. Thick (0.5-0.7 μm) sections were cut on glass knives and collected on Formvar-coated slot grids stained with uranylacetate and lead citrate and examined with the AEI EM7 1.2 MV HVEM in Albany, NY at 1000 kV.

Download Full-text