The expressible fluid of fish fillets. VIII.—cell damage in slow freezing

This is a preliminary report of electron probe analysis of rabbit portal-anterior mesenteric vein (PAMV) smooth muscle cryosectioned without fixation or cryoprotection. The instrumentation and method of electron probe quantitation used (1) and our initial results with cardiac (2) and skeletal (3) muscle have been presented elsewhere.In preparations depolarized with high K (K2SO4) solution, significant calcium peaks were detected over the sarcoplasmic reticulum (Fig 1 and 2) and the continuous perinuclear space. In some of the fibers there were also significant (up to 200 mM/kg dry wt) calcium peaks over the mitochondria. However, in smooth muscle that was not depolarized, high mitochondrial Ca was found in fibers that also contained elevated Na and low K (Fig 3). Therefore, the possibility that these Ca-loaded mitochondria are indicative of cell damage remains to be ruled out.

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Histological evaluation of cochlear hair cell damage from noise-induced hearing loss in chinchillas

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100159217 ◽

1990 ◽

Vol 48 (3) ◽

pp. 332-333

Author(s):

R.V. Harrison ◽

R.J. Mount ◽

P. White ◽

N. Fukushima

Keyword(s):

Hair Cell ◽

Evoked Potential ◽

Cell Damage ◽

Acoustic Trauma ◽

Histological Evaluation ◽

Cell Integrity ◽

Functional Changes ◽

Cochlear Hair Cell ◽

Left And Right ◽

Contralateral Ear

In studies which attempt to define the influence of various factors on recovery of hair cell integrity after acoustic trauma, an experimental and a control ear which initially have equal degrees of damage are required. With in a group of animals receiving an identical level of acoustic trauma there is more symmetry between the ears of each individual, in respect to function, than between animals. Figure 1 illustrates this, left and right cochlear evoked potential (CAP) audiograms are shown for two chinchillas receiving identical trauma. For this reason the contralateral ear is used as control.To compliment such functional evaluations we have devised a scoring system, based on the condition of hair cell stereocilia as revealed by scanning electron microscopy, which permits total stereociliar damage to be expressed numerically. This quantification permits correlation of the degree of structural pathology with functional changes. In this paper wereport experiments to verify the symmetry of stereociliar integrity between two ears, both for normal (non-exposed) animals and chinchillas in which each ear has received identical noise trauma.

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Osmolarity and Artificial Cell Damage in Ischemic Myocardium

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100158868 ◽

1990 ◽

Vol 48 (3) ◽

pp. 262-263

Author(s):

Richard Montione ◽

Muhammad Ashraf

Keyword(s):

Cell Damage ◽

Uranyl Acetate ◽

Tissue Preservation ◽

Rat Myocardium ◽

En Bloc ◽

Cellular Changes ◽

Artificial Cell ◽

Cacodylate Buffer ◽

Isotonic Buffer ◽

Perfusion Fixation

Osmolarity of a fixative vehicle has long been known to have an effect on the tissue preservation. An increase in tissue osmolarity occurs in ischemia-damaged tissue and affects the morphology. In this study, we examined cellular changes in ischemic rat myocardium induced by varying fixative toxicity.Rats were sacrificed by decapitation and the hearts immediately removed and retrogradily perfused through the aorta with anoxic Kurbs-Henseleit medium. Hearts were then placed in a bag with a small amount of medium at 37°C for 90 minutes. Hearts were perfusion-fixed using 2% glutaraldehyde in 0.1 M cacodylate buffer pH -7.3 at three osmolarities. The isotonic buffer was adjusted to 311 mOsm/kg using D-manitol. Hypertonic buffers were adjusted to 375 and 400 mOsm/kg. One-half hour after perfusion fixation, the hearts were sliced and cut into small blocks and allowed to fix overnight at 4°C. Blocks were post fixed in osmium, en bloc stained in uranyl acetate, dehydrated in ethanol and embedded in Spurr medium.

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