Solid surface vitrification: A promising approach for testis tissue preservation

Cryobiology ◽  
2020 ◽  
Vol 97 ◽  
pp. 302
Author(s):  
Tanushree Patra ◽  
Mukesh Kumar Gupta
Author(s):  
J. G. Adams ◽  
M. M. Campbell ◽  
H. Thomas ◽  
J. J. Ghldonl

Since the introduction of epoxy resins as embedding material for electron microscopy, the list of new formulations and variations of widely accepted mixtures has grown rapidly. Described here is a resin system utilizing Maraglas 655, Dow D.E.R. 732, DDSA, and BDMA, which is a variation of the mixtures of Lockwood and Erlandson. In the development of the mixture, the Maraglas and the Dow resins were tested in 3 different volumetric proportions, 6:4, 7:3, and 8:2. Cutting qualities and characteristics of stability in the electron beam and image contrast were evaluated for these epoxy mixtures with anhydride (DDSA) to epoxy ratios of 0.4, 0.55, and 0.7. Each mixture was polymerized overnight at 60°C with 2% and 3% BDMA.Although the differences among the test resins were slight in terms of cutting ease, general tissue preservation, and stability in the beam, the 7:3 Maraglas to D.E.R. 732 ratio at an anhydride to epoxy ratio of 0.55 polymerized with 3% BDMA proved to be most consistent. The resulting plastic is relatively hard and somewhat brittle which necessitates trimming and facing the block slowly and cautiously to avoid chipping. Sections up to about 2 microns in thickness can be cut and stained with any of several light microscope stains and excellent quality light photomicrographs can be taken of such sections (Fig. 1).


Author(s):  
Richard Montione ◽  
Muhammad Ashraf

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.


2020 ◽  
pp. 49-56
Author(s):  
Vitaly V. Volkov ◽  
Michael A. Suslin ◽  
Jamil U. Dumbolov

One of the conditions for ensuring the safety of air transport operation is the quality of aviation fuel refueled in aircraft. Fuel quality control is a multi-parameter task that includes monitoring the free moisture content. Regulatory documents establish the content of free water no more than 0.0015% by weight. It is developed a direct electrometric microwave resonance method for controlling free moisture in aviation fuels, which consists in changing the shape of the water drops by pressing them on a solid surface inside a cylindrical cavity resonator. This can dramatically increase dielectric losses. Analytical and experimental analysis of the proposed method is carried out. The control range from 0,5 to 30 μl of absolute volume of moisture in aviation fuels with a maximum error of not morethan 25 % is justified. The sensitivity of the proposed method for monitoring microwave losses in free moisture drops transformed into a thin layer by pressing is an order of magnitude greater than the sensitivity of the method for monitoring microwave losses in moisture drops on a solid surface in a resonator. The proposed method can be used as a basis for the development of devices for monitoring the free moisture of aviation fuels in the conditions of the airfield and laboratory. The direction of development of the method is shown.


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