Correlation of high-pressure arc heater results

The ultrastructure of cells suspended in physiological fluids or cell culture media can only be studied if the living processes are stopped while the cells remain in suspension. Attachment of living cells to carrier surfaces to facilitate further processing for electron microscopy produces a rapid reorganization of cell structure eradicating most traces of the structures present when the cells were in suspension. The structure of cells in suspension can be immobilized by either chemical fixation or, much faster, by rapid freezing (cryo-immobilization). The fixation speed is particularly important in studies of cell surface reorganization over time. High pressure freezing provides conditions where specimens up to 500μm thick can be frozen in milliseconds without ice crystal damage. This volume is sufficient for cells to remain in suspension until frozen. However, special procedures are needed to assure that the unattached cells are not lost during subsequent processing for LVSEM or HVEM using freeze-substitution or freeze drying. We recently developed such a procedure.

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Prolonged Fixation Studies For Spaceflight

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100072101 ◽

1973 ◽

Vol 31 ◽

pp. 332-333

Author(s):

Robert Corbett ◽

Delbert E. Philpott ◽

Sam Black

Keyword(s):

High Pressure ◽

Living Systems ◽

Prolonged Storage ◽

Time Intervals ◽

Early Signs ◽

Large Numbers

Observation of subtle or early signs of change in spaceflight induced alterations on living systems require precise methods of sampling. In-flight analysis would be preferable but constraints of time, equipment, personnel and cost dictate the necessity for prolonged storage before retrieval. Because of this, various tissues have been stored in fixatives and combinations of fixatives and observed at various time intervals. High pressure and the effect of buffer alone have also been tried.Of the various tissues embedded, muscle, cartilage and liver, liver has been the most extensively studied because it contains large numbers of organelles common to all tissues (Fig. 1).

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Cryosectioning plant roots prepared by high-pressure freezing

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100127748 ◽

1987 ◽

Vol 45 ◽

pp. 662-663

Author(s):

R.E. Crang ◽

M. Mueller ◽

K. Zierold

Keyword(s):

High Pressure ◽

Cell Walls ◽

Plant Tissues ◽

Ice Crystal ◽

High Pressure Freezing ◽

Vitreous State ◽

Radial Depth ◽

Irregular Topography ◽

Considerable Depth ◽

Conventional Freezing

Obtaining frozen-hydrated sections of plant tissues for electron microscopy and microanalysis has been considered difficult, if not impossible, due primarily to the considerable depth of effective freezing in the tissues which would be required. The greatest depth of vitreous freezing is generally considered to be only 15-20 μm in animal specimens. Plant cells are often much larger in diameter and, if several cells are required to be intact, ice crystal damage can be expected to be so severe as to prevent successful cryoultramicrotomy. The very nature of cell walls, intercellular air spaces, irregular topography, and large vacuoles often make it impractical to use immersion, metal-mirror, or jet freezing techniques for botanical material.However, it has been proposed that high-pressure freezing (HPF) may offer an alternative to the more conventional freezing techniques, inasmuch as non-cryoprotected specimens may be frozen in a vitreous, or near-vitreous state, to a radial depth of at least 0.5 mm.

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