Staining Paraffin Sections with Protargol 3. The Optimum pH for Reduction. 4. A Two-Hour Staining Method

Davenport, H.; McArthur, Janet W.; Bruesch, S. R.; Holland, Beulah F.

doi:10.3109/10520293909110288

Cited by 46 publications

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“…Less attention will be paid the changes which occur in the heart muscle and other organs of chronically deficient pigeons with signs of cardiac failure. The osmic acid and chlorateosmic acid methods have been used for studying the myelin sheaths, and Davenport's rapid silver (protargol) method (16) has been depended upon to detect changes in the axis cylinder. It will be pointed out elsewhere (17) that, contrary to prevailing opinion, these methods detect very early degenerative changes in peripheral nerves (in the rat no later than 8 hours after the sciatic nerve has been sectioned).…”

Section: (From the Department Of Pathology The Harvard Medical Schoomentioning

confidence: 99%

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Avian Thiamin Deficiency

Swank

1940

Journal of Experimental Medicine

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PLAXES 31 TO 34Many investigators (1-6) have demonstrated that peripheral nerve fibers of animals deficient in vitamin B1 (thiamin) degenerate, and some (3,5,6) have noted similar but less marked changes in control starved animals. Other investigators (7-10) found no, or few, degenerating nerve fibers in thiamin deficient rats, chicks, or other animals, and almost all have expressed the opinion that the symptoms of thiamin deficiency cannot be explained on the basis of observable pathological lesions. Non-specific vacuolation and shrinkage of nerve cells in various parts of the central nervous system have been seen frequently in both thiamin deficient and starved control animals, but in a recent review Vedder (11) pointed out that marked changes, the appearance of which suggested exhaustion, were to be found in ganglion cells of the nervous system, especially in the lumbosacral region of the spinal cord?I t is apparent from these contradictory reports that the histopathology of thiamin deficiency is unsettled. Some investigators do not believe that degenerative lesions occur in thiamin deficient animals; others feel that the lesions that have been described are due to an absence of other factors from the diet, or to starvation; whereas those who did find degenerative lesions in this condition appear to be uncertain as to their exact nature. Finally, one is led to the conclusion that the lesions which were observed in starved animals are of the same indefinite type, e.g., degeneration of myelin or demyelinization, as have been observed in thiamin deficient animals. With one exception (7), the clinical symptoms that were observed in thiamin deficient animals were distinct from anything observed in starved or otherwise deficient animals.The general conclusion that a degeneration of myelin (or demyelinization) occurs in thiamin deficient and starved animals has been confirmed by the polarized light technique (12, 13), and by the same method it has been pointed out that many more degenerating nerve fibers are to be found in chronic than in acutely deficient rats (13).In a recent study of starvation in rats ~ marked alterations were observed in the

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Section: (From the Department Of Pathology The Harvard Medical Schoomentioning

confidence: 99%

“…The sections through the cervical enlargement of the spinal cord including the dorsal and sympathetic ganglia and attached peripheral nerve, were stained as well by the rapid silver (protargol) method of Davenport (16).…”

Section: Histopathological Techniquementioning

confidence: 99%

Avian Thiamin Deficiency

Swank

1940

Journal of Experimental Medicine

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“…The reducer is a sodium sulphite-sodium bisulphite, buffered, amidol solution with a pH of 6-5 to 7 5 which is the optimum for amidol (Davenport, McArthur, and Bruesch, 1939). We found this formula to be very superior to any other reducer, e.g., hydroquinone, pyrogallic acid, formol, etc., or their combinations.…”

Section: Indirect Albumen (Silver-gold) Methodsmentioning

confidence: 91%

Methods of Impregnating Neurofibrillar Substance on Slides After Imbedding

Mihálik¹

1950

Journal of Neurology, Neurosurgery & Psychiatry

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This paper describes three new methods for impregnating neurofibrillar substance in sections prepared from imbedded material and then mounted on slides. The first two methods are simpler than any previous slide methods for, after the silver bath, the usual reduction is left out and the silver is replaced directly by gold. All these methods can be modified to some extent for selective impregnation of connective tissue fibres (Fig. 4) simply by adding two drops of concentrated ammonia per 100 ml. to the colloidal silver bath, but a discussion of the impregnation of the connective tissue fibres is not included in this paper.I. Direct Albumen (Silver-Gold) Method By using 0-2 to 0-5% egg albumen and 2% silver nitrate in aqueous solution (a colloidal silver bath) it is possible to impregnate the neurofibrillar substance of the central nervous system on slides of imbedded and sectioned materials fixed in a variety of ways. The slides are kept in the silver colloid bath for five days at 370 C. or for eight days at room temperature, then gilded for 10 minutes by dropping 022% aqueous gold chloride solution onto the slides, and subsequently reduced in a 0 2% aqueous oxalic acid solution for an hour. Between the steps of the technique, especially between the silver colloid and gold, the slides have to be well washed with distilled water.This method is extremely simple: over-or underimpregnation or reduction is impossible; the results are selective, free of precipitate, and constant; it works even after decalcification of the material to be impregnated by trichloro-acetic acid.Fixation.

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“…Longer washes, of up to several minutes, offer no advantages and slowly remove unreduced silver from the tissues, making the stain paler. A similar short rinse after gold toning also gives cleaner preparations (Davenport et al 1939). Other washes do not seem critical.…”

Section: Rinsesmentioning

confidence: 95%

“…Nevertheless, the quality of successful results was good enough to warrant an investigation of the factors controlling the stain. The Bodian method has been widely used on vertebrate material and the effects of varying some of the conditions of staining have been reported (Bodian 1937;Davenport et al 1938Davenport et al , 1939Davenport et al , 1947Holmes 1943), but little was known of its use on invertebrates. Power devised the double-impregnation procedure for insect tissue-the central nervous system of Drosophila melanogaster-and subsequent workers (Hess 1958;Pipa et al 1959;Chen and Chen 1969) have used it with little alteration, though Rogoff (1946) stained mosquito brains with only one impregnation.…”

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confidence: 99%

Silver Staining of Insect Central Nervous Systems by the Bodian Protargol Method

Gregory

1970

Acta Zoologica

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Improved fixation of ganglia of the central nervous system of Periplaneta americana and Schistocerca gregaria for silver staining by Power's (1943) modification of the Bodian protargol method is given by alcoholic Bouin aged for at least 40 days at 60° C. During impregnation of sections, increased copper and decreased pH give paler staining, more selective for nerve fibres. Prolonging impregnation from 24 to 48 hours weakens the stain and decreases selectivity. The intensity of the stain depends chiefly upon the amount of unreduced (developable) silver combined with the tissues; selectivity is determined mainly by the number and distribution of the reduced silver particles (‘nuclei’). In development, increased sodium sulphite gives more differentiation, increased hydroquinone gives less. Optimum developer composition depends upon impregnation, and thick sections need more differentiation than thinner ones. Within limits, change in one of the factors that control staining can be balanced by changes in others, but by suitable adjustment of the conditions the result can be varied from almost total staining of nerve fibres, for general neuroanatomy, to highly selective staining for tracing individual fibres.

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Staining Paraffin Sections with Protargol 3. The Optimum pH for Reduction. 4. A Two-Hour Staining Method

Cited by 46 publications

References 1 publication

Avian Thiamin Deficiency

Avian Thiamin Deficiency

Methods of Impregnating Neurofibrillar Substance on Slides After Imbedding

Silver Staining of Insect Central Nervous Systems by the Bodian Protargol Method

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