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.

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 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.

Periodic acid-thiocarbohydrazide-silver methenamine(PATS) reaction: An improved silver methodology for light and electron microscopy

1984 ◽  
Vol 42 ◽  
pp. 264-265 ◽  
Author(s):  
B. Giammara ◽  
T. Romaine ◽  
W. Ambrose ◽  
J. Hanker
Keyword(s):  
Electron Microscopy ◽  
Osmium Tetroxide ◽  
Periodic Acid ◽  
Patent Application ◽  
Light And Electron Microscopy ◽  
Basement Membranes ◽  
Full Description ◽  
Periodic Acid Schiff ◽  
Reticular Fibers ◽  
Pas Reaction

Many variations of the periodic acid-Schiff(PAS) reaction have been utilized for electron microscopy based on the Gomori periodic acid-silver methenamine reaction (1) or the periodic acid-thiocarbohydrazide-osmium tetroxide(PATCO) reaction (2,3). These reactions are widely employed and have been very useful for the demonstration of one or more biomacromolecules or structures such as glycogen, basement membranes, reticular fibers or lipopolysaccharide. However, these reactions have various drawbacks such as complexity of methodology, ability to stain only a limited number of these components, or lack of adaptability for both light and electron microscopy. Our newly devised PATS reaction is relatively easy to perform. A full description of the details must await the outcome of a pending patent application. It consists essentially of a stepwise treatment of the sample with periodic acid, thiocarbohydrazide(TCH) and silver methenamine.

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 Apparent failure of 2-hydroxy-3-naphthoic acid hydrazide to block the periodic acid-Schiff reactivity of sialic acids. Implications for the PANFOPAS method.

1983 ◽  
Vol 31 (9) ◽  
pp. 1142-1144 ◽  
Author(s):  
P E Reid ◽  
C F Culling
Keyword(s):  
Sialic Acid ◽  
Periodic Acid ◽  
Acid Hydrazide ◽  
Sialic Acids ◽  
Side Chain ◽  
Substitution Pattern ◽  
Periodic Acid Schiff ◽  
Naphthoic Acid ◽  
Vicinal Diols ◽  
Anionic Groups

Histochemical studies of the mechanism of staining of the periodic acid/2,-hydroxy-3-naphthoic acid hydrazide/Fast Black B/saponification/periodic acid-Schiff (PANFOPAS) method indicate that while 2-hydroxy-3-naphthoic acid hydrazide blocks the periodate engendered Schiff reactivity of vicinal diols unassociated with anionic groups, it fails to block such reactivity in sialic acid residues. It is suggested, therefore, that the positive Schiff staining observed following application of the PANFOPAS method to colonic epithelial mucins may be due to sialic acids without side chain substituents (or which are substituted at C7 or C9) in addition to sialic acids substituted at C8 (or which are di- or trisubstituted). Consequently the PANFOPAS method cannot be used to study the side chain substitution pattern of the sialic acids of epithelial mucins.

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