Staining of Blood Cells with Periodic Acid/Salicyloyl Hydrazide (PA-SH)

Blood ◽  
1966 ◽  
Vol 28 (5) ◽  
pp. 674-682 ◽  
Author(s):  
J. BURNS ◽  
P. B. NEAME
Keyword(s):  
Guinea Pig ◽  
Acute Leukemia ◽  
Blood Cells ◽  
Periodic Acid ◽  
Acid Oxidation ◽  
Periodic Acid Schiff ◽  
Type Method ◽  
Tissue Sections ◽  
Periodic Acid Oxidation ◽  
Schiff Reaction

Abstract Stoward1 conjugated acidified solutions of salicyloyl hydrazide with the dialdehydes formed from the periodic acid oxidation of vicinal glycols in guinea pig tissue sections. The method has now been utilized, with minor modification, to demonstrate glycogen in blood and marrow cells, and it has been compared with the periodic acid-Schiff reaction and with a fluorescent acriflavine Schiff-type method. It is felt that the PA-SH method will replace the existing Schiff-type fluorescent methods and that it will prove to be a useful technic to aid in the diagnosis of blood conditions, such as acute leukemia, where PAS positivity is known to occur.

scholarly journals Determination of the glycogen content in single neutrophil leukocytes using a micromodel of leukocyte glycogen.

1975 ◽  
Vol 23 (1) ◽  
pp. 59-64 ◽  
Author(s):  
G Gahrton ◽  
I Olsson ◽  
A Dahlqvist
Keyword(s):  
Periodic Acid ◽  
Liver Glycogen ◽  
Mean Value ◽  
Acid Oxidation ◽  
Periodic Acid Schiff ◽  
Periodic Acid Oxidation ◽  
Normal Human ◽  
Schiff Reaction ◽  
Neutrophil Leukocytes

The kinetics of the periodic acid oxidation as part of the periodic acid-Schiff reaction was studied by combined microinterferometry and microspectrophotometry in micromodel systems of liver glycogen and leukocyte glycogen as well as in neutrophil leukocytes. The initial formation of Schiff-positive chromogens was more rapid in neutrophil leukocytes than in liver or leukocyte glycogen. The chromogen formation was, however, practically complete within 60 min in both neutrophil leukocytes and leukocyte glycogen, but this did not appear to be the case in liver glycogen. Differences in the rate of chromogen formation may depend on various factors such as differences in the source and treatment of the glycogen. The complete periodic acid-Schiff reaction appears to be a measure of the glycogen amount in neutrophil leukocytes and the microdroplet system of leukocyte glycogen is considered to be an appropriate model for the estimation of the glycogen amount in single neutrophil leukocytes. A mean value of 13.3 10-12 g glycogen per normal human neutrophil was found.

The Histochemical Demonstration of Keratin by Methods involving Selective Oxidation

1951 ◽  
Vol s3-92 (20) ◽  
pp. 393-402
Author(s):  
A. G. EVERSON PEARSE
Keyword(s):  
Periodic Acid ◽  
Cobalt Nitrate ◽  
Histochemical Demonstration ◽  
Performic Acid ◽  
Cysteic Acid ◽  
Acid Oxidation ◽  
Reaction Products ◽  
Periodic Acid Schiff ◽  
Periodic Acid Oxidation ◽  
Schiff Reaction

1. Oxidation of tissues with performic acid gives rise to histochemically detectable reaction products particularly in two classes of material. These are keratin and lipoids of the phosphatide class. 2. Three methods have been evolved for visualizing the effect of performic acid on cystine-containing structures; two of these (performic acid/Schiff and perfoiTnic acid/cobalt nitrate) also record the effect on lipoids. 3. An attempt has been made to elucidate the chemistry of the reactions and it is suggested that oxidation of cystine in the tissues gives rise not only to cysteic acid (alanine-beta-sulphonic) but to another acid (alanine-beta-sulphinic). The latter is responsible for the positive reaction with Schiff's solution. 4. The Schiff reaction with performic acid oxidized lipoids is due to the formation of substances giving the reactions of aldehydes. It is possible that similar groups may be produced from lipoid molecules by periodic acid oxidation and that these and not polysaccharides (1.2 glycols) are responsible for the periodic acid-Schiff reaction in such cases.

scholarly journals Observations on the Foam Cell in Thalassemia

Blood ◽  
1960 ◽  
Vol 16 (1) ◽  
pp. 1039-1044 ◽  
Author(s):  
P. C. SEN GUPTA ◽  
J. B. CHATTERJEA ◽  
A. M. MUKHERJEE ◽  
ANJALI CHATTERJI
Keyword(s):  
Sulfuric Acid ◽  
Foam Cell ◽  
Histochemical Study ◽  
Periodic Acid ◽  
Periodic Acid Schiff ◽  
Tissue Sections ◽  
Histiocytic Cell ◽  
Acid Type ◽  
Schiff Reaction ◽  
Red Coloration

Abstract Histologic and histochemical study of the foam cell in the spleen in thalassemia has shown that the foamy appearance is due to the accumulation of an acidic mucopolysaccharide of the chondroitin sulfuric acid type in the cytoplasm of this histiocytic cell. This polysaccharide shows intense red coloration with the periodic acid Schiff reaction, which is thus of great value in demonstrating the foam cell in tissue sections. The possible sources of this mucopolysaccharide have been discussed.

scholarly journals The Cytochemistry of Acute Leukemia: Observations on Glycogen and Neutral Fat in Bone Marrow Aspirates

Blood ◽  
1969 ◽  
Vol 33 (2) ◽  
pp. 341-347 ◽  
Author(s):  
JOHN M. BENNETT ◽  
THOMAS F. DUTCHER
Keyword(s):  
Bone Marrow ◽  
Acute Leukemia ◽  
Periodic Acid ◽  
Periodic Acid Schiff ◽  
Cytochemical Study ◽  
Blast Cells ◽  
Schiff Reaction ◽  
Granulocytic Leukemia ◽  
Oil Red O

Abstract A cytochemical study of 54 cases of acute leukemia utilizing the periodic acid-Schiff reaction for glycogen and Oil Red O for neutral fat was performed on air dried bone marrow smears. The majority of leukemic lymphoblasts revealed significant amounts of glycogen and neutral fat. "Blast" cells in granulocytic leukemia were devoid of glycogen and of neutral fat. The recommendation is made that both of these histochemical stains be employed to attempt to resolve the problem of specific cellular identification in acute leukemia.

Microspectrofluorometry of a periodic acid-Schiff reaction in blood cells

1969 ◽  
Vol 56 (1) ◽  
pp. 59-68 ◽  
Author(s):  
X. Yataghanas ◽  
G. Gahrton ◽  
B. Thorell
Keyword(s):  
Blood Cells ◽  
Periodic Acid ◽  
Periodic Acid Schiff ◽  
Schiff Reaction

scholarly journals HISTOCHEMICAL DEMONSTRATION OF SOME OXIDIZED MACROMOLECULES WITH THIOCARBOHYDRAZIDE (TCH) OR THIOSEMICARBAZIDE (TSC) AND OSMIUM TETROXIDE

1965 ◽  
Vol 13 (8) ◽  
pp. 629-639 ◽  
Author(s):  
ARNOLD M. SELIGMAN ◽  
JACOB S. HANKER ◽  
HANNAH WASSERKRUG ◽  
HERMINE DMOCHOWSKI ◽  
LIONEL KATZOFF
Keyword(s):  
Osmium Tetroxide ◽  
Periodic Acid ◽  
Acid Hydrazide ◽  
Histochemical Demonstration ◽  
Basement Membranes ◽  
Acid Oxidation ◽  
Cell Nuclei ◽  
Periodic Acid Schiff ◽  
Naphthoic Acid ◽  
Periodic Acid Oxidation

Thiocarbohydrazide (TCH) and thiosemicarbazide (TSC) have been introduced as osmiophilic reagents for demonstrating aldehyde-containing macromolecules, produced by periodic acid oxidation of tissue sections. Because of the remarkable properties of osmium black, excellent pigment qualities are provided for light microscopy and the advantages of an amorphous, nonvolatile, electron-opaque end product are available for electron microscopy. The present communication describes the method and gives the scope of the histochemical reaction. Thiocarbohydrazide is the more reactive of the two reagents and is capable of demonstrating a wider variety of oxidized macromolecules than does thiosemicarbazide. The periodic acid-thiocarbohydrazide-osmium tetroxide method (PATCO method) was compared with those using thiosemicarbazide (PATO method), periodic acid-Schiff (PAS method), and with the method involving 3-hydroxy-2-naphthoic acid hydrazide (NAH) followed by coupling with fast blue B (PA-NAH-FBB method). All four methods demonstrated certain active aldehydes equally well, such as those produced by periodic acid oxidation of glycogen, mucopolysaccharide of gut and mucoprotein of Descemet's membranes of the cornea. The stain with the PATO method was somewhat weaker than the other three methods for glycoprotein of the cuticular border of gut and of the brush border of kidney tubules, and colloid of the thyroid gland. The PATO method was even less effective in demonstrating mucoprotein of basement membranes in kidney and no reaction for collagen and reticulin was noted. The PATCO method was almost as effective with basement membranes in kidney as the PAS method, but was not equal to the PAS method in staining collagen and reticulin. Furthermore, the aldehyde produced on Feulgen hydrolysis of deoxyribonucleic acid (DNA) of cell nuclei was not demonstrable with either PATO or PATCO methods its contrast to positive results with the PAS and PA-NAH-FBB methods. This suggests that TSC and TCH have less reactive hydrazino groups than NAH. Results with the electron microscope will be published separately.

scholarly journals The presence of ethylenic bonds and vic-glycol groups in neuromelanin and lipofuscin in the human brain.

1983 ◽  
Vol 31 (7) ◽  
pp. 849-858 ◽  
Author(s):  
H Barden
Keyword(s):  
Substantia Nigra ◽  
Oxalic Acid ◽  
Human Brain ◽  
Inferior Olive ◽  
Periodic Acid ◽  
Performic Acid ◽  
Acid Oxidation ◽  
Periodic Acid Oxidation ◽  
Schiff Reaction ◽  
Subsequent Staining

Through use of oxidation and blockading reactions, chemical group precursors of aldehyde demonstrable with Schiff reaction staining were identified in the soluble lipid-free lipofuscin component of neuromelanin of human substantia nigra and in lipofuscin of human inferior olive. Aldehyde generation was implied by moderate staining after bleaching neuromelanin and oxidizing lipofuscin with potassium permanganate followed by oxalic acid. Confirmation of aldehyde generation was achieved when diminished staining followed a sulfite addition blockade obtained by replacing oxalic acid with metabisulfite or bisulfite as well as by condensation blockades obtained with phenylhydrazine or aniline without replacing oxalic acid. Vic-glycol precursors of aldehyde were demonstrated in both pigments when acetylation or bromination preceded permanganate-oxalic acid and staining was unequivocally diminished only after acetylation. Vic-glycols were also demonstrated in lipofuscin by diminished staining when acetylation preceded periodic acid oxidation. Ethylenic precursors of aldehyde were suggested in performic acid-bleached neuromelanin when the minimal staining that followed this peracid's generation of Schiff reaction-negative dihydroxy groups became greatly intensified following an additional oxidation with periodic acid. This additional oxidation converted the dihydroxys to Schiff reaction-positive aldehyde. Ethylenes in lipofuscin were indicated when bromination before performic acid reduced subsequent staining.

scholarly journals HISTOCHEMICAL ACYLATION OF ALDEHYDES PRODUCED BY PERIODIC ACID OXIDATION

1966 ◽  
Vol 14 (7) ◽  
pp. 529-537 ◽  
Author(s):  
R. D. LILLIE
Keyword(s):  
Acetic Acid ◽  
Sulfuric Acid ◽  
Acetic Anhydride ◽  
Glacial Acetic Acid ◽  
Periodic Acid ◽  
Potassium Hydroxide Solution ◽  
Acid Oxidation ◽  
Enol Acetate ◽  
Periodic Acid Oxidation ◽  
Schiff Reaction

Aldehydes produced in tissue sections by periodic acid oxidation are readily acetylated or benzoylated so as to weaken or completely prevent the Schiff and other chromogenic reactions. The reactivity of acylated aldehydes to Schiff reagent is promptly restored by saponification in alcoholic potassium hydroxide solution (10-20 min, 1% KOH, 70% ethanol). Benzoylation with 5-10% benzoyl chloride in pyridine gave the promptest, most complete and consistent aldehyde blockade, which was somewhat inferior on addition of 0.5% sulfuric acid. Pyridine acetic anhydride mixtures, 25, 40 and 50% gave partial to complete blockade. Addition of 0.25-0.5% sulfuric acid did not give consistent effects. Acetic anhydride at 60°C gave partial blockade at ½-5 hr, on addition of 0.01-0.25% sulfuric acid total or subtotal blockade was achieved. Acetylation in alcohol gave inferior resutlts. Use of 25% acetic anhydride ims glacial acetic acid gave inferior results; addition of 0.25% sulfuric acid produced total to subtotal blockade at 4-5 hr. Glacial acetic acid was without appreciable blockade effect. Sulfation in 10% and 25% H2SO4/glacial acetic acid failed to blockade aldehydes. Experiments with the peracetic acid Schiff reaction for ethylene groups indicate that some proportion of enol monobenzoate may be formed, and that with the various acetylation techniques a smaller or negligible proportion of enol acetate is formed. Acetic anhydride with 0.25% H2S04 at 60° appears to form only aldehyde diacetate. Experiments with the bromination silver techniques did not give satisfactory results.

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