Utilization of Pre-Rigor Pork Loin for the Manufacture of Low-Salt Pork Loin and Rapid Freezing Effect

Bismuth (Bi) stains nucleoproteins (NPs) by interacting with available amino and primary phosphate groups. These two staining mechanisms are distinguishable by glutaraldehyde crosslinking (Fig. 1,2).Isolated mouse liver nuclei, extracted with salt and acid solutions, fixed in either formaldehyde (form.) or gl utaraldehyde (glut.) and stained with Bi, were viewed to determine the effect of the extractions on Bi stainina. Solubilized NPs were analyzed by SDS-polyacrylamide gel electrophoresis.Extraction with 0.14 M salt does not change the Bi staining characteristics (Fig. 3). 0.34 M salt reduces nucleolar (Nu) staining but has no effect on interchromatinic (IC) staining (Fig. 4). Proteins responsible for Nu and glut.- insensitive IC staining are removed when nuclei are extracted with 0.6 M salt (Fig. 5, 6). Low salt and acid extraction prevents Bi-Nu staining but has no effect on IC staining (Fig. 7). When nuclei are extracted with 0.6 M salt followed by low salt and acid, all Bi-staining components are removed (Fig. 8).

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Ultra-Rapid Freezing of Isolated Skeletal Muscle Fibers

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100080316 ◽

1977 ◽

Vol 35 ◽

pp. 576-577

Author(s):

Joachim R. Sommer ◽

Nancy R. Wallace

Keyword(s):

Skeletal Muscle ◽

Sarcoplasmic Reticulum ◽

Granular Material ◽

Nuclear Envelope ◽

Intracellular Calcium ◽

Ruthenium Red ◽

Threshold Concentration ◽

Rapid Freezing ◽

Frog Skeletal Muscle ◽

Electrical Isolation

After Howell (1) had shown that ruthenium red treatment of fixed frog skeletal muscle caused collapse of the intermediate cisternae of the sarcoplasmic reticulum (SR), forming a pentalaminate structure by obi iterating the SR lumen, we demonstrated that the phenomenon involves the entire SR including the nuclear envelope and that it also occurs after treatment with other cations, including calcium (2,3,4).From these observations we have formulated a hypothesis which states that intracellular calcium taken up by the SR at the end of contraction causes the M rete to collapse at a certain threshold concentration as the first step in a subsequent centrifugal zippering of the free SR toward the junctional SR (JSR). This would cause a) bulk transport of SR contents, such as calcium and granular material (4) into the JSR and, b) electrical isolation of the free SR from the JSR.

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Cryogenic atomic force microscopy

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100084570 ◽

1991 ◽

Vol 49 ◽

pp. 54-55

Author(s):

K. A. Fisher ◽

M. G. L. Gustafsson ◽

M. B. Shattuck ◽

J. Clarke

Keyword(s):

Room Temperature ◽

Rapid Freezing ◽

Cryogenic Temperatures ◽

Soft Or ◽

Electrically Conductive ◽

Force Microscopy ◽

Flexible Molecules ◽

Advantages And Disadvantages ◽

Atomic Force ◽

Chemical Perturbation

The atomic force microscope (AFM) is capable of imaging electrically conductive and non-conductive surfaces at atomic resolution. When used to image biological samples, however, lateral resolution is often limited to nanometer levels, due primarily to AFM tip/sample interactions. Several approaches to immobilize and stabilize soft or flexible molecules for AFM have been examined, notably, tethering coating, and freezing. Although each approach has its advantages and disadvantages, rapid freezing techniques have the special advantage of avoiding chemical perturbation, and minimizing physical disruption of the sample. Scanning with an AFM at cryogenic temperatures has the potential to image frozen biomolecules at high resolution. We have constructed a force microscope capable of operating immersed in liquid n-pentane and have tested its performance at room temperature with carbon and metal-coated samples, and at 143° K with uncoated ferritin and purple membrane (PM).

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Freeze-substitution of rye grass and ragweed pollen grains

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100160984 ◽

1990 ◽

Vol 48 (3) ◽

pp. 686-687

Author(s):

Liza B. Martinez ◽

Susan M. Wick

Keyword(s):

Cell Function ◽

Pollen Grains ◽

Freeze Substitution ◽

Cell Types ◽

Rapid Freezing ◽

Rye Grass ◽

Acrylic Resins ◽

Allergenic Proteins ◽

Cold Embedding ◽

Structural Preservation

Rapid freezing and freeze-substitution have been employed as alternatives to chemical fixation because of the improved structural preservation obtained in various cell types. This has been attributed to biomolecular immobilization derived from the extremely rapid arrest of cell function. These methods allow the elimination of conventionally used fixatives, which may have denaturing or “masking” effects on proteins. Thus, this makes them ideal techniques for immunocytochemistry, in which preservation of both ultrastructure and antigenicity are important. These procedures are also compatible with cold embedding acrylic resins which are known to increase sensitivity in immunolabelling.This study reveals how rapid freezing and freeze-substitution may prove to be useful in the study of the mobile allergenic proteins of rye grass and ragweed. Most studies have relied on the use of osmium tetroxide to achieve the necessary ultrastructural detail in pollen whereas those that omitted it have had to contend with poor overall preservation.

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Ultrastructural and immunocytochemical studies of the rat hypothalamo-neurohypophysial system processed by rapid freezing and freeze-substitution fixation

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100161989 ◽

1990 ◽

Vol 48 (3) ◽

pp. 884-885

Author(s):

Seiji Shioda ◽

Yasumitsu Nakai ◽

Atsushi Ichikawa ◽

Hidehiko Ochiai ◽

Nobuko Naito

Keyword(s):

Osmium Tetroxide ◽

Freeze Substitution ◽

Ultrastructural Localization ◽

Wistar Rat ◽

Neurosecretory Cells ◽

Rapid Freezing ◽

Sodium Metaperiodate ◽

Tissue Sections ◽

Ultrathin Sections ◽

Electron Microscopes

The ultrastructure of neurosecretory cells and glia cells in the supraoptic nucleus (SON) of the hypothalamus and the neurohypophysis (PN) was studied after rapid freezing followed by substituion fixation. Also, the ultrastructural localization of vasopressin (VP) or its carrier protein neurophys in II (NPII) in the SON and PN was demonstrated by using a post-embedding immunoco1loidal gold staining method on the tissue sections processed by rapid freezing and freeze-substitution fixation.Adult male Wistar rat hypothalamus and pituitary gland were quenched by smashing against a copper block surface precooled with liquid helium and freeze-substituted in 3% osmium tetroxide-acetone solutions kept at -80°C for 36-48h. After substituion fixation, the tissue blocks were warmed up to room temperature, washed in acetone and then embedded in an Epon-Araldite mixture. Ultrathin sections mounted on 200 mesh nickel grids were immersed in saturated sodium metaperiodate and then incubated in each of the following solutions: 1 % egg albumin in phosphate buffer, VP or NPII (1/1000-1/5000) antiserum 24h at 4°C, 3) colloidal gold solution (1/20) 1h at 20°C. The sections were washed with distilled waterand dried, then stained with uranylacetate and lead citrate and examined with Hitachi HU-12A and H-800 electron microscopes.

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Rapid Freezing of Freeze-Etch Specimens

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s1431927600003275 ◽

1980 ◽

Vol 38 ◽

pp. 752-755

Author(s):

A. Elgsaeter ◽

T. Espevik ◽

G. Kopstad

Keyword(s):

Low Temperature ◽

Metal Surface ◽

Relative Velocity ◽

Thermal Contact ◽

High Rate ◽

Rapid Freezing ◽

Temperature Decrease ◽

Freeze Etching

The importance of a high rate of temperature decrease (“rapid freezing”) when freezing specimens for freeze-etching has long been recognized1. The two basic methods for achieving rapid freezing are: 1) dropping the specimen onto a metal surface at low temperature, 2) bringing the specimen instantaneously into thermal contact with a liquid at low temperature and subsequently maintaining a high relative velocity between the liquid and the specimen. Over the last couple of years the first method has received strong renewed interest, particularily as the result of a series of important studies by Heuser and coworkers 2,3. In this paper we will compare these two freezing methods theoretically and experimentally.

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Surfactant Impurities Can Explain the Jones-Ray Effect

10.26434/chemrxiv.5732976 ◽

2018 ◽

Author(s):

Timothy Duignan ◽

Marcel Baer ◽

Christopher Mundy

Keyword(s):

Surface Tension ◽

Electrostatic Potential ◽

Driving Forces ◽

Salt Water ◽

Fundamental Property ◽

Apparent Contradiction ◽

Low Salt ◽

Air Water Interface ◽

Added Salt ◽

Effect Of Surface

<div> <p> </p><div> <div> <div> <p>The surface tension of dilute salt water is a fundamental property that is crucial to understanding the complexity of many aqueous phase processes. Small ions are known to be repelled from the air-water surface leading to an increase in the surface tension in accordance with the Gibbs adsorption isotherm. The Jones-Ray effect refers to the observation that at extremely low salt concentration the surface tension decreases in apparent contradiction with thermodynamics. Determining the mechanism that is responsible for this Jones-Ray effect is important for theoretically predicting the distribution of ions near surfaces. Here we show that this surface tension decrease can be explained by surfactant impurities in water that create a substantial negative electrostatic potential at the air-water interface. This potential strongly attracts positive cations in water to the interface lowering the surface tension and thus explaining the signature of the Jones-Ray effect. At higher salt concentrations, this electrostatic potential is screened by the added salt reducing the magnitude of this effect. The effect of surface curvature on this behavior is also examined and the implications for unexplained bubble phenomena is discussed. This work suggests that the purity standards for water may be inadequate and that the interactions between ions with background impurities are important to incorporate into our understanding of the driving forces that give rise to the speciation of ions at interfaces. </p> </div> </div> </div> </div>

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