Variations in the Gram Staining Results Caused by Air Moisture

Tucker, F. L.; Bartholomew, J. W.

doi:10.3109/10520296209117724

Cited by 4 publications

(5 citation statements)

References 3 publications

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“…Cell-wall porosity and thickness, however, could be important in that a thin cell wall with large water-filled "pores" (gram-negative) would present a rather favorable permeation situation as compared with thick cell walls with smaller "pore" spaces (gram-positive). In this respect, it is interesting that yeast cells are much slower to decolorize than B. subtilis cells (Tucker and Bartholomew, 1962), and they have thicker cell walls, smaller "pores," and a lower percentage of water content in the cell-wall material (Gerhardt and Judge, 1964). It is hoped that the "pore" size and water-content determinations of Gerhardt and Judge will be extended to include a representative series of cell walls from gram-negative as well as gram-positive organisms.…”

Section: Discussionmentioning

confidence: 99%

Analysis of the Mechanism of Gram Differentiation by Use of a Filter-Paper Chromatographic Technique

1965

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Analysis of the mechanism of Gram differentiation by use of a filter-paper chromatographic technique. J. Bacteriol. 90:766-777. 1965.-Data are presented which demonstrate that the mechanism of gram-positivity could not be due solely to factors such as a single, specific gram-positive substrate, specific affinities of crystal violet for certain cellular components, a specific crystal violet-iodine-substrate complex, or to any specific characteristic of the dye, iodine, or solvent molecules. Ruptured cells of gram-positive organisms stain gram-negatively when subjected to a standard Gram-stain procedure. However, when stained fragments of broken cells were deposited in thick layers on the surface of filter-paper strips and exposed to decolorizers, the rate of dye release correlated with the Gram characteristic of the intact cell. Therefore, the intact cell in itself is not an absolute requirement for Gram differentiation. The data are interpreted as indicating that the mechanism of Gram differentiation primarily involves the rate of permeation of molecules (dye, iodine, solvent) through the interstitial spaces of cell-wall material.

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Section: Discussionmentioning

confidence: 99%

Analysis of the Mechanism of Gram Differentiation by Use of a Filter-Paper Chromatographic Technique

1965

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“…However, since it was desired to have a more quantitative comparison of decolorization characteristics than that used by Chelton & Jones, a Gram procedure described by Bartholomew (1962) also was used. Briefly, the latter involved the substitution of Hucker's crystal violet for 0.5 yo methyl violet 6B, the use of 95 yo (v/v) ethanol in water as a decolorizer rather than 90 yo (v/v) ethanol in water and the use of a quantitative decolorization procedure.…”

Section: Methodsmentioning

confidence: 99%

“…The question which arises then, is what criteria can be used to evaluate the validity of results obtained with any single Gram procedure. The answer is simple if one is only concerned with intact cells and if one closely follows the ordinarily recommended Gram procedures (Bartholomew, 1962). However, if one studies ruptured organisms, or used heat in staining, or changes the decolorizer or decolorization procedure, and if a t the same time one sticks to a single decolorization time, then all that emerges blue is not necessarily Gram-positive.…”

Section: Effect Of Moisture On Gram Differentiation 263mentioning

confidence: 99%

“…Such quantitative determinations are not possible using the usual Gram procedures with a single decolorization time. If all of the known variables of the Gram stain procedure were kept constant (Bartholomew, 1962), then a single decolorization time known to give valid differentiations could be used with a fair degree of confidence. If, however, the known variables are changed, and if determinations of the Gram nature of an unknown substrate under different conditions are to be made, then the only valid experimental procedure is to use some form of a quantitative Gram procedure.…”

Section: Effect Of Moisture On Gram Differentiation 263mentioning

confidence: 99%

“…The mechanism of Gram-positivity was interpreted, then, as a combination of the presence of a Gram-positive component, and selective permeability to ethanol of the intact cell envelopes. Bartholomew (1962) showed that intact cells, both Gram-positive and Gramnegative, were influenced by wet and dry Gram procedures, as well as by the water content of the decolorizer, in respect to the rate of decolorization. This presents the possibility that the differences observed by Chelton & Jones for ruptured cells can be explained without resorting to the concept of a Gram-positive component.…”

Section: Introductionmentioning

confidence: 99%

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The Effect of Moisture on Gram Differentiation, and its Relation to Proposed Gram-Positive Substrates in Yeast and Other Organisms

Bartholomew¹,

Tucker²,

Finkelstein³

1964

Journal of General Microbiology

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SUMMARYWet and dry Gram staining procedures have different characteristics in respect to their rates of decolorization. It has been proposed that this difference is due to a Gram-positive substrate which dissociates (becomes Gram-negative) in the presence of water. However, the differences between wet and dry Gram procedures can also be explained on the hypothesis that water influences the rate of solvent permeation through cell envelopes. Since no direct proof exists for the reality of the proposed Gram-positive substrate, and since evidence is presented here which cannot be explained by such a hypothesis, it is felt that the solvent permeation concept should be considered seriously as an explanation for the decolorization differences observed between wet and dry Gram procedures.

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An alcohol-soluble Schiff's reagent: A histochemical application of the complex between Schiff's reagent and phosphotungstic acid

Bedi

Horobin

1976

Histochemistry

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A schedule for staining partially hydrated PAS-positive structures using non-aqueous solutions has been devised. Tissues are dewaxed, taken down to 70% alcohol, oxidised for 10 min in a 1% w/v alcoholic solution of periodic acid, treated with an alcoholic solution of phosphotungstic acid-Schiff reagent complex (PTA-Schiff reagent) for 25 min, washed in alcohol, cleared in xylene and mounted in a synthetic medium. The PTA-Schiff reagent complex prepared from de Tomasi Schiff reagent by precipitation with PTA may be stored in the deep freeze for many months and dissolved freshly in alcohol for use. The PTA-Schiff reagent used as above allows staining of highly water soluble materials such as dextran. From blocking and digestion studies the mode of action seems similar to de Tomasi Schiff reagent. The partial hydration of the tissues prior to reaction was found to be essential for effective staining.

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Variations in the Gram Staining Results Caused by Air Moisture

Cited by 4 publications

References 3 publications

Analysis of the Mechanism of Gram Differentiation by Use of a Filter-Paper Chromatographic Technique

Analysis of the Mechanism of Gram Differentiation by Use of a Filter-Paper Chromatographic Technique

The Effect of Moisture on Gram Differentiation, and its Relation to Proposed Gram-Positive Substrates in Yeast and Other Organisms

An alcohol-soluble Schiff's reagent: A histochemical application of the complex between Schiff's reagent and phosphotungstic acid

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