The Vital Staining of Amoeba Proteus

1963 ◽  
Vol s3-104 (68) ◽  
pp. 445-458
Author(s):  
JENNIFER M. BYRNE
Keyword(s):  
Nile Blue ◽  
Vital Staining ◽  
Neutral Red ◽  
Amoeba Proteus ◽  
Methyl Green ◽  
Pronounced Increase ◽  
Brilliant Cresyl Blue ◽  
Cresyl Blue ◽  
Vital Dyes ◽  
Food Vacuoles

The effect of keeping Amoeba proteus in dilute basic dye solutions was studied. It was found that Nile blue, neutral red, and neutral violet in particular, and also brilliant cresyl blue, methylene blue, Bismarck brown, thionin, toluidine blue, and azures A and B act as vital dyes, while at comparable molarities crystal violet, dahlia, safranin, methyl green, Janus green, and Victoria blue are lethal, and do not produce any staining until after death. Azure C, basic fuchsin, and particularly pyronine G are relatively harmless, but produce no vital staining. All the vital dyes stain the food vacuoles, and all produce small, darkly stained granules in colourless vacuoles in the cytoplasm. The latter do not exist in the unstained amoeba. Some of the dyes colour vacuoles around the crystals. These crystal vacuoles also seem to be induced. A few of the dyes colour the spherical refractive bodies, which are at least in part phospholipid. All the basic dyes used with the possible exception of azure C, methyl green, and pyronine G attach to the external membrane of A. proteus in an orientated manner, as shown by the increase in birefringence of the external membrane induced by these dyes. It is particularly those dyes that act as vital dyes that produce a very pronounced increase in the birefringence of the external membrane.

The indicator system of the Sabin-Feldman phenomenon in Toxoplasma

Parasitology ◽  
1960 ◽  
Vol 50 (1-2) ◽  
pp. 13-20
Author(s):  
C. Kulasiri
Keyword(s):  
Toluidine Blue ◽  
Nile Blue ◽  
Neutral Red ◽  
Indicator System ◽  
Alkaline Solutions ◽  
Methyl Green ◽  
Celestine Blue ◽  
Cresyl Blue ◽  
Azure B ◽  
Saline Solutions

Azure A, azure B and Nile blue in both neutral and alkaline solutions were found to be suitable for use as indicator dyes in the Sabin-Feldman test. Toluidine blue, brilliant cresyl blue and thionine were also suitable as neutral solutions. Azure C, phloxine, phloxine B, neutral red, methyl green, pyronine Y, eosin and celestine blue were found to be unsuitable. Azure B and toluidine blue in saline solutions were serviceable up to 5 months after preparation. The significance of these findings is discussed.

scholarly journals STUDIES OF THE INNER AND OUTER PROTOPLASMIC SURFACES OF LARGE PLANT CELLS

1944 ◽  
Vol 28 (1) ◽  
pp. 17-22 ◽  
Author(s):  
W. J. V. Osterhout
Keyword(s):  
Plant Cells ◽  
Good Deal ◽  
Neutral Red ◽  
Central Vacuole ◽  
Normal Position ◽  
Brilliant Cresyl Blue ◽  
Large Plant ◽  
Cresyl Blue ◽  
Mechanical Disturbance ◽  
Outer Protoplasmic Surface

The vacuolar surface of Nitella is covered with a non-aqueous film too thin to be visible as a separate membrane. The motion of the protoplasm may subject this film to a good deal of mechanical disturbance. Apparently this does not rupture the film for no dye escapes into the protoplasm as the result of such disturbance when the vacuolar sap is deeply stained with neutral red or brilliant cresyl blue. When the deeply stained central vacuole breaks up into several smaller vacuoles, leaving the outer protoplasmic surface in its normal position, there is no evidence of the escape of dye into the protoplasm through the film surrounding the vacuole.

A simple technique for the preservation of vital dyes in fixed and sectioned embryos

Development ◽  
1966 ◽  
Vol 15 (2) ◽  
pp. 131-132
Author(s):  
J. Pertusa
Keyword(s):  
Methylene Blue ◽  
Ethyl Alcohol ◽  
Toluidine Blue ◽  
Trypan Blue ◽  
Simple Technique ◽  
Nile Blue ◽  
Neutral Red ◽  
The Other ◽  
Serial Sectioning ◽  
Vital Dyes

In embryological work using vital dyes it is highly desirable to be able to study the distribution of the dyes in fixed material, whether examined in toto or after embedding in paraffin and serial sectioning. However, both fixation and dehydration present problems for the preservation of colour in vitally stained cells. Some fixatives preserve some dyes but, so far as I am aware, none will preserve all the vital dyes in common use. On the other hand, ethyl alcohol destroys or dissolves all vital dyes and its use in dehydration is thus undesirable. Among the fixatives that have been proposed are those of Golowin (1902), Mitamura (1923), Parat & Painlevé (1925), and Tchéou Tai Chuin (1930) for neutral red; that of Izquierdo (1955) for toluidine blue; that of Gérard (1925) for Trypan blue; that of Turchini (1919) for methylene blue; that of Lehmann (1929) for Nile blue.

VITAL DYES FOR MARKING LIVING ELATERID LARVAE

1963 ◽  
Vol 41 (6) ◽  
pp. 991-996 ◽  
Author(s):  
R. Y. Zacharuk
Keyword(s):  
Rhodamine B ◽  
Fat Body ◽  
Nile Blue ◽  
Methyl Groups ◽  
Molecular Configuration ◽  
Brilliant Cresyl Blue ◽  
Cresyl Blue ◽  
Sudan Black B ◽  
Fat Bodies ◽  
Selective Storage

The following dyes appear to be suitable as internal vital markers for living wireworms: Nile blue sulphate, rhodamine B, Sudan black B, brilliant cresyl blue, acridine orange N, coriphosphine O, and brilliant fat scarlet. All are assimilated quickly after ingestion with food, are stored in the fat body for extended periods, can be seen distinctly through the integument, and have no apparent toxic effects. Their selective storage in the fat bodies appears to be a product of their content of the fat-soluble amino-ethyl or amino-methyl groups, and of a specific type of molecular configuration. These dyes will undoubtedly also be useful as vital markers for other insects, particularly those with thin, transparent integuments.

Memoirs: A Study of the Yolk (Y-granules) of the Male Germ-cells

1936 ◽  
Vol s2-78 (311) ◽  
pp. 513-531
Author(s):  
J. A. MULIYIL
Keyword(s):  
Fatty Acids ◽  
Fatty Acid ◽  
Germ Cells ◽  
Nile Blue ◽  
Saturated Fatty Acids ◽  
Vital Staining ◽  
Neutral Red ◽  
Osmic Acid ◽  
Male Germ Cells ◽  
Staining Techniques

1. Recent researches have established that the Y-granules exist as a fundamental part of the constitution of the male germ-cells of many animals, vertebrates as well as invertebrates. 2. Prior to the application of vital staining techniques to cytological problems, Saccocirrus was the only animal in which these granules were known to exist. 3. The fixatives generally used by cytologists, especially those containing acetic acid and other fat solvents, are not indicated for studying the Y-granules. Vital staining techniques offer the best method for their study. 4. Underfeeding and starvation of the larvae of Agrotis segetum and Abraxas grossulariata revealed that these granules are products of normal metabolic activity, and that their appearance and disappearance depend on the general physiological status of the animal. 5. In normally fed larvae the Y-granules were invariably present, and responded to the vital dye within a few minutes of its application. The germ-cells of underfed larvae showed a steady decrease in their Y-granule contents, as did starved larvae up to a certain stage. When starvation was prolonged the granules disappeared from the cell. 6. The changes produced by underfeeding and starvation in the cell, both in the Y-granule content and the colloidal state of the cytoplasm, were decidedly more pronounced in the spermatocytes than in the spermatids. In very advanced stages of the cell scarcely any change was perceptible. 7. The chemical composition of the granules was determined by studying their reactions to certain vital dyes and fixatives. Neutral red, brilliant Cresyl blue, and Nile-blue sulphate were successfully used to stain the granules, Nile-blue sulphate being particularly satisfactory. This stain is specific for fats and substances chemically allied to fats. 8. Y-granules are composed of either fatty acids or a mixture of fatty acids and neutral fat, because they always stain blue with Nile-blue sulphate. 9. Fixatives containing osmic acid failed to stain the granules except in rare cases when they appeared brown in the preparations. As a rule they appeared a shade of pale grey. As this is a reaction for saturated fatty acids it is believed that in the majority of cases where in fixed preparations the granules are seldom visible, they are composed of saturated fatty acids. 10. The fatty nature of the material was confirmed by fixing testis smears in formalin vapour, and staining them with Herxheimer's solution of acetone and scarlet B. If the preparations are counterstained in methylene blue the Y-granules are clearly seen stained pink--a reaction indicating the fatty acid nature of the granules. 11. An examination of the ovaries of Abraxas grossul ariata, Gryllus domesticus, and Lithobius forficatus, supra vitally stained in neutral red, revealed in everycase a juxtanuclear aggregate of neutral red staining granules. With the growth of the oocytes the granules wandered into the cytoplasm and gradually developed into small spheres; their substance simultaneously underwent a chemical change. By the aid of Nile-blue sulphate it was possible to ascertain that the fatty acid contents of the granules gradually changed into fat, or rather into fatty yolk. 12. The behaviour of the granules in the oocytes suggest that the Y-granules in the spermatocytes are only abortive yolk granules having no function under normal conditions. But a gradual reduction in their number, observed in several spermatocytes when the animals were starved, suggests that under exceptional circumstances there is a possibility of the granules exercising some sort of storage function.

Memoirs: The Cytology of Amoeba Proteus ‘Y’ and the Effects of Large and Small Centrifugal Forces

1938 ◽  
Vol s2-80 (320) ◽  
pp. 601-634
Author(s):  
B. N. SINGH
Keyword(s):  
Golgi Apparatus ◽  
Nile Blue ◽  
Cyst Formation ◽  
Neutral Red ◽  
Amoeba Proteus ◽  
Contractile Vacuole ◽  
Osmic Acid ◽  
Culture Solution ◽  
Food Material ◽  
Sudan Iv

1. When Amoeba proteus is subjected to high centrifugal force most of the cytoplasmic bodies are thrown out of the cell, so this work was done with the ordinary electrical centrifuge. 2. The stratification of the various cytoplasmic components according to their specific gravity is as follows: the contractile vacuole and the fat, being the lightest, occupy the centripetal position; then there is a layer of cytoplasm followed by mitochondria, neutral-red bodies, crystals, and nutritive spheres. The nucleus occupies a position in between the crystals and the nutritive spheres (Text-fig. 2). 3. The redistribution of the various cytoplasmic components takes place within a few minutes after amoebae have been centrifuged by the electrical centrifuge. Ultra-centrifuged organisms kept in culture solution remain rounded for 10-15 days, and no cyst formation takes place. The crystals and nutritive spheres are reformed; the former seem to be the products of excretion formed by the metabolic activity of the cell. 4. The nutritive spheres contain glycogen as reserve food material, and give positive tests for glycogen with iodine and Best's carmine. 5. There is no evidence that the bodies which stain with neutral red are the homologue of the metazoan Golgi apparatus, although they are pre-existing bodies in Amoeba proteus . The contractile vacuole does not blacken even after prolonged osmication. No certain homologue of the Golgi apparatus was found in Amoeba proteus. 6. Fat and glycogen are two distinct types of storage material present in Amoeba proteus. The former is very well seen with osmic acid, Sudan IV, and Nile blue tests.

Examining bivalve fecundity: oocyte viability revealed by Neutral Red vital staining

2021 ◽  
Author(s):  
Peter G. Beninger ◽  
Daphné Chérel ◽  
Lucie Kessler
Keyword(s):  
Vital Staining ◽  
Neutral Red
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