scholarly journals AN EXPERIMENTAL COMPARISON OF DIFFERENT CRITERIA OF DEATH IN YEAST

1933 ◽  
Vol 16 (4) ◽  
pp. 579-592 ◽  
Author(s):  
Otto Rahn ◽  
Margaret Noble Barnes

Different criteria of death have been compared experimentally and quantitatively. Pure cultures of a yeast have been subjected to adverse conditions, and the number of dead cells, judged by different tests, has been determined in successive time intervals. The yeast cultures were exposed to heat, to HgCl2, to ultraviolet light, and to x-rays. In each case, the cells lost the power of reproduction (measured by the plate count) most rapidly. The loss of fermentation (measured by the CO2 pressure) was less rapid. Still slower was the change in staining reaction with methylene blue, and the loss of selective permeability of the plasma membrane (measured by the percentage of cells staining with Congo red). Slowest of all was the coagulation of protoplasm as observed in the dark-field. In the case of death by heat or by HgCl2, the rate of loss of reproduction was about twice as rapid as that of the loss of fermentation, about three times that of the loss of semipermeability, and about forty times as large as the rate of coagulation. With ultraviolet light and with x-rays, these ratios were decidedly different. The technique employed does not permit the conclusion that any one criterion of death is the prerequisite for other criteria. It does not appear probable that loss of reproduction is the prerequisite for loss of fermentation or of semipermeability because the ratios of the velocities of these processes are not the same with all causes of death. There is no evidence that cells may show certain criteria of death immediately after exposure, and recover later.

1996 ◽  
Vol 2 (2) ◽  
pp. 53-62 ◽  
Author(s):  
Henry N. Chapman ◽  
Jenny Fu ◽  
Chris Jacobsen ◽  
Shawn Williams

The methods of immunolabeling make visible the presence of specific antigens, proteins, genetic sequences, or functions of a cell. In this paper we present examples of imaging immunolabels in a scanning transmission x-ray microscope using the novel method of dark-field contrast. Colloidal gold, or silver-enhanced colloidal gold, is used as a label, which strongly scatters x-rays. This leads to a high-contrast dark-field image of the label and reduced radiation dose to the specimen. The x-ray images are compared with electron micrographs of the same labeled, unsectioned, whole cell. It is verified that the dark-field x-ray signal is primarily due to the label and the bright-field x-ray signal, showing absorption due to carbon, is largely unaffected by the label. The label can be well visualized even when it is embedded in or laying behind dense material, such as the cell nucleus. The resolution of the images is measured to be 60 nm, without the need for computer processing. This figure includes the x-ray microscope resolution and the accuracy of the label positioning. The technique should be particularly useful for the study of relatively thick (up to 10 μm), wet, or frozen hydrated specimens.


1995 ◽  
Vol 58 (3) ◽  
pp. 301-304 ◽  
Author(s):  
YOGA PANDYA ◽  
FRED F. JEWETT ◽  
DALLAS G. HOOVER

Saccharomyces cerevisiae ATCC 2373 and Zygosaccharomyces bailii ATCC 36947 were exposed to hydrostatic pressures ranging from 1,500 to 3,000 atmospheres for 10, 20 and 30 min in 0.1 M citrate buffer at pH 3.0, 4.0 and 5.0 at 25 and 45°C. Inactivation of inoculated yeast cultures was achieved in spaghetti sauce with meat at 25°C with 3,000 atmospheres for 10 min and also at 45°C and 2,500 atmospheres for 10 min. Viable counts were determined on potato dextrose agar (PDA) incubated at 30°C for 48 h. Pressure-induced injury was demonstrated by plate count differential between PDA and PDA supplemented with glucose (PDAG). A reduction of 7-log10 cycles colony forming units (CFU)/ml was seen for both strains at 3,000 atmospheres for 10 min at 25°C at all pH levels and at 2,250 atmospheres, pH 5.0 for 20 min at 45°C. At 2,000 atmospheres, pH 3.0 for 30 min, the increase in temperature from 25 to 45°C increased the inactivation of yeast by 6-log10 cycles. Lowering the pH from 5.0 to 3.0 enhanced lethality up to 2-log10 cycles at 2,250 atmospheres, 25°C for 30 min. Injury was most apparent at exposure parameters that produced 3- to 5-log10 cycle reductions on PDA. This was achieved (99% injury) at 2,250 atmospheres, 25°C for 30 min. These data indicate that mild heat and acidity contribute to the effectiveness of the inactivation and injury of yeast by high hydrostatic pressure (HHP).


1968 ◽  
Vol 31 (6) ◽  
pp. 177-179 ◽  
Author(s):  
W. C. Frazier ◽  
D. F. Gneiser

Directions are given for the 8- to 12-hr membrane filter-colony technique for the estimation of numbers of viable bacteria in rinsings or swabbings from the surfaces of foods or equipment. Tests on 6 species of bacteria often found in foods demonstrated good recovery of organisms from pure cultures or mixtures of 2 or 3 cultures. Tests with dairy rinses and fresh green beans and sweet com gave counts almost as high as by the 48-hr plate, count. Results with frozen or blanched and frozen beans and corn and with chlorinated flume waters were unsatisfactory.


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