John A Kiernan

The University of Western Ontario | UWO · Department of Anatomy and Cell Biology

We investigated physicochemical characteristics of dye lots sold as “alcian blue” using the Biological Stain Commission (BSC) precipitation test, differential scanning calorimetry, high performance liquid chromatography, thin layer chromatography and UV/visible spectroscopy. Four blue phthalocyanine dyes were detected in 11 commercial dye lots. The...

Eriochrome cyanine R (C.I. 43820, Mordant blue 3), also known as chromoxane cyanine R and solochrome cyanine R, has been used as a biological stain since 1957. In conjunction with ferric ions, it provides selective blue coloration of the nuclei of cells in methods procedurally similar to commonly used progressive or regressive hemalum (aluminum-hem...

Alcian blue dyes are copper phthalocyanines with a variety of cationic side chains; they are useful for staining carbohydrate polyanions while avoiding staining of nucleic acids. The properties of the original alcian blue and of similar dyes with published chemical structures are reviewed here. Variation among samples submitted to the Biological St...

Previous investigators have disagreed about whether hemalum stains DNA or its associated nucleoproteins. I review here the literature and describe new experiments in an attempt to resolve the controversy. Hemalum solutions, which contain aluminum ions and hematein, are routinely used to stain nuclei. A solution containing 16 Al³⁺ ions for each hema...

Lipids are a heterogeneous group of substances characterized by their solubility in organic solvents and insolubility in water. Lipids can be found as normal components of different tissues and organs, and they can be affected by several pathological conditions. The histochemical identification of lipids plays an important role in histopathological...

. For the 5th fifth edition (2015) - ISBN 9781907904325 - see this link: http://www.scionpublishing.com/titles/215-9781907904325-histological-and-histochemical-methods-fifth-edition . The first two editions (1981, 1990) were published by Pergamon Press, The third editon (1999) was published by Butterworths. The fourth (2008) and fifth (2015) editio...

Only primates have temporal lobes, which are largest in man, accommodating 17% of the cerebral cortex and including areas with auditory, olfactory, vestibular, visual and linguistic functions. The hippocampal formation, on the medial side of the lobe, includes the parahippocampal gyrus, subiculum, hippocampus, dentate gyrus, and associated white ma...

A technique has been developed whereby osmium tetroxide (OSO4) is reclaimed from solutions used as fixatives in electron microscopy. The osmium compounds in the collected residues are oxidized to OSO4, which is then extracted into carbon tetrachloride. Osmium dioxide is precipitated with alcohol, recovered by filtration and dried. This is a safe an...

Sirius red F3B (CI 35780, Direct red 80) is a polyazo dye used principally in staining methods for collagen and amyloid. For certification by the Biological Stain Commission, a sample of the dye must exhibit an absorption spectrum of characteristic shape with a maximum at 528-529 nm, a small shoulder near 500 nm and narrow peaks at 372, 281-282 and...

Kiernan, J. A. 2010. On chemical reactions and staining mechanisms. Connection (Dako Inc. Scientific Magazine). Vol. 14, pp. 127-136. [What is the difference between argentaffin and argyrophilic reactiuons? (p. 127-128). How does pH influence the staining mechanism of Schiff's reagent in biological tissue samples? (p. 129-131). What is Giemsa’s sta...

In recent decades, graduate students and other research workers have received little formal education in the principles of microtechnique. Consequently they often encounter difficulties with simple staining methods and make mistakes as the result of insufficient understanding of the principles of microtechnique. This article describes a half-course...

The origins of repeated hematoxylin shortages are outlined. Lack of integration in the hematoxy-lin trade exacerbates the problems inherent in using a natural product. Separate corporations are engaged in tree growth and harvesting, dye extraction, processing of extracts to yield hematoxylin, and formulation and sale of hematoxylin staining solutio...

The origins of repeated hematoxylin shortages are outlined. Lack of integration in the hematoxylin trade exacerbates the problems inherent in using a natural product. Separate corporations are engaged in tree growth and harvesting, dye extraction, processing of extracts to yield hematoxylin, and formulation and sale of hematoxylin staining solution...

A numerical scoring system is presented for evaluating structural fixation of certain mammalian tissues for light microscopy. Small pieces of rat's kidney and brain, tissues for which artifacts of fixation are well documented, were fixed in various fluids. Random code numbers hid their identities, and paraffin sections were stained to show nuclear...

In the three earlier editions of News from the Biological Stain Commission (BSC), under the heading of "Regulatory affairs," the BSC's International Affairs Committee reported on the work of Technical Committee 212, Clinical Laboratory Testing and in Vitro Diagnostic Test Systems of the International Standards Organization (ISO/TC 212) and its work...

In the three earlier editions of News from the Biological Stain Commission (BSC), under the heading of “Regulatory affairs,” the BSC's International Affairs Committee reported on the work of Technical Committee 212, Clinical Laboratory Testing and in Vitro Diagnostic Test Systems of the International Standards Organization (ISO/TC 212) and its work...

INTRODUCTION For the study of microscopic anatomy and of pathological material, it is usual to stain sections of tissue in such a way as to impart a dark color to the nuclei of cells and a lighter, contrasting color to the cytoplasm and extracellular structures. Nuclear stains, including cationic, anionic, and metal complexing dyes, are considered...

INTRODUCTION For the study of microscopic anatomy and of pathological material, it is usual to stain sections of tissue in such a way as to impart a dark color to the nuclei of cells and a lighter, contrasting color to the cytoplasm and extracellular structures. In most of the general “oversight” methods used in histology, a blue, purple, or black...

This review summarizes the function, structure, and chemistry of the myelin sheaths of axons: discusses the mechanisms of several staining techniques using inorganic compounds, dyes, and antibodies; and provides technical instructions for 13 staining methods for normal and degenerating myelin. Myelin has alternating hydrophobic and hydrophilic laye...

Indoxyl esters and glycosides are useful chromogenic substrates for detecting enzyme activities in histochemistry, biochemistry and bacteriology. The chemical reactions exploited in the laboratory are similar to those that generate indigoid dyes from indoxyl-beta-d-glucoside and isatans (in certain plants), indoxyl sulfate (in urine), and 6-bromo-2...

Carmine is one of the original dyes certified by the Biological Stain Commission (BSC). Until now it has lacked both an assay procedure for dye content and a means to positively identify the dye. The methods for testing carmine in the laboratory of the BSC have been revised to include spectrophotometric examination at pH 12.5-12.6 to determine that...

Nuclear fast red (CI 60760), also known as Kernechtrot, is commonly used in conjunction with an excess of aluminum ions as a red nuclear counterstain following histochemical procedures that yield blue products. The dye has also been used as a histochemical and colorimetric reagent for calcium. Unsatisfactory samples of nuclear fast red are encounte...

Isolation of well-preserved pure cell populations is a prerequisite for sound studies of the molecular basis of any tissue-based biological phenomenon. This article reviews current methods for obtaining anatomically specific signals from molecules isolated from tissues, a basic requirement for productive linking of phenotype and genotype. The quali...

Isolation of well-preserved pure cell populations is a prerequisite for sound studies of the molecular basis of pancreatic malignancy and other biological phenomena. This chapter reviews current methods for obtaining anatomically specific signals from molecules isolated from tissues, a basic requirement for productive linking of phenotype and genot...

Isolation of well-preserved pure cell populations is a prerequisite for sound studies of the molecular basis of pancreatic malignancy and other biological phenomena. This chapter reviews current methods for obtaining anatomically specific signals from molecules isolated from tissues, a basic requirement for productive linking of phenotype and genot...

Hexazonium pararosaniline is a valuable reagent that has been used in enzyme activity histochemistry for 50 years. It is an aqueous solution containing the tris-diazonium ion derived from pararosaniline, an aminotriarylmethane dye, and it contains an excess of nitrous acid that was not consumed in the diazotization reaction. Other investigators hav...

Myogenesis is regulated by the MyoD class of myogenic regulatory factors (MRFs). These basic helix-loop-helix transcription factors dimerize with E proteins to bind conserved E-box sequences in the promoter regions of muscle-specific genes. Perhaps due to their expression in a wide array of tissues, the specific interactions of E proteins with diff...

The origins of repeated hematoxylin shortages are outlined. Lack of integration in the hematoxy-lin trade exacerbates the problems inherent in using a natural product. Separate corporations are engaged in tree growth and harvesting, dye extraction, processing of extracts to yield hematoxylin, and formulation and sale of hematoxylin staining solutio...

Here we describe a method for gross staining of gray matter in slices of formaldehyde-fixed human brain. After protection of white matter with 4% phenol at 60 degrees C for 5 min followed by a cold water wash, the gray matter was stained for 10-15 min at 20-25 degrees C with 1% aqueous copper(II) phthalocyanine tetrasulfonic acid tetrasodium salt (...

A classification of dyes and other colorants is proposed, based on the chemical features responsible for their visibility and generally consonant with the writings of modern color chemists. The scheme differs in several respects from that of the Colour Index (CI), but it retains some traditional small groups of dyes that include biological stains....

Osmium tetroxide is indeed wonderful stuff. Osmium is a rare element, so disposal of used solutions should consist of recycling, not dumping, even though osmium compounds are not considered environmentally hazardous (Smith et al. , 1978 Trace Metal in the Environment , vol 4. Ann Arbor Science Publishers). The colorless and soluble toxic tetroxide....

Aldehydes are the most commonly used fixatives. They serve to stabilize the fine structural details of cells and tissues prior to examination by light or electron microscopy. Research workers, technicians, pathologists and others who regularly use aldehyde fixatives frequently do not appreciate the nature and properties of these compounds or the re...

Cerebral aneurysms are composed principally of collagen, a birefringent protein which is responsible for withstanding the forces of blood pressure. The known correlation between collagen birefringence and its mechanics provides the basis for using polarizing microscopy to evaluate the strength of collagen, layer by layer across the aneurysmal wall....

3,3'-Diaminobenzidine (DAB) is widely used as a chromogen for visualization of horseradish peroxidase activity in neuroanatomical tracing experiments and in immunohistochemistry. The product of the enzymatically catalyzed oxidation of DAB by hydrogen peroxide is brown and nonfluorescent. In frozen sections of formaldehyde fixed rat and mouse brain...

Chemical dehydration can be accomplished using 2,2-dimethoxypropane (DMP). In the presence of an acid catalyst, this liquid reacts with water generating methanol and acetone as products. Although DMP is more expensive per milliliter than ethanol and other solvents used for dehydration, it is an economical alternative because a much smaller volume i...

The requirements for such a medium are that it won't deposit crystals when it evaporates and that it won't be incapable of evaporating. The second condition probably excludes anything containing glycerol. The first is more difficult. A polymer such as gum, polyvinylpyrollidone or gelatin might fiil the bill, especially in the presence of a sugar th...

Incluye bibliografía e índice

Anionic dyes, notably acid fuchsine, strongly stain the nuclei and cytoplasm of neurons severely damaged by injury or disease. We provide detailed instructions for staining nervous tissue with toluidine blue and acid fuchsine for optimal demonstration of injured neurons. Degeneration was induced in the hippocampus of the mouse by systemic administr...

Paraffin sections are usually rehydrated before staining. It is possible to apply aqueous dye solutions without first removing the wax. Staining then occurs more slowly, and only if the embedding medium has not melted or become unduly soft after cutting. To avoid this problem, sections are flattened on water no hotter than 45 C and dried overnight...

Endothelium, choroidal epithelium, and arachnoid exclude plasma proteins from most parts of the mammalian central nervous system (CNS). Nerve roots, in contrast, have permeable capillaries and permeable pia-arachnoid sheaths. Diffusion of plasma proteins into the cerebrospinal fluid is probably prevented by slow bulk flow along a pressure gradient...

The dyes commonly used in histological staining provide little information about the innervation of skeletal muscle because they reveal neither the terminal parts of axons nor the postsynaptic structures of the motor end-plate. Silver methods, which reveal axons of all types, are best applied to thick (50–100μm) frozen sections. The chemical reacti...

Ubiquitin targets proteins for attack by certain proteolytic enzymes, but the ubiquitinated cytoplasmic inclusions seen in some chronic neurodegenerative diseases may indicate the occurrence of reparative rather than destructive metabolic events. We have examined the production of ubiquitin in motor neurons of the rat's left hypoglossal nucleus aft...

Neuronal degeneration in the precentral gyrus alone cannot account for the occurrence of spastic paresis in motor neuron diseases. To look for more extensive cortical atrophy we measured MRIs of the upper parts of the frontal and parietal lobes in 11 sporadic cases of classical amyotrophic lateral sclerosis (ALS), eight patients with primary latera...

We have attempted to resolve apparently conflicting observations of previous investigators regarding the penetration of proteins into peripheral ganglia and nerves of the rat. Horseradish peroxidase, which is largely cleared from the blood and extracellular fluids in less than 30 min, entered all the extracellular spaces of ganglia, including the c...

Autoantibodies, a suggested cause of motor neurone degeneration in amyotrophic lateral sclerosis (ALS), were sought immunohistochemically after applying diluted sera to sections of normal human spinal cord. Serum from a case of paraneoplastic motor neurone disease provided a positive control, giving strong staining of motor neurones for IgG but not...

In amyotrophic lateral sclerosis, motor neurons in the spinal cord and brainstem shrink before they die. In 12 cases of sporadic amyotrophic lateral sclerosis, and in 11 control subjects, we have measured the neurons in the second sacral segment and the hypoglossal nucleus, and have calculated a ‘shape index’ that shows the convexity or concavity o...

Primary lateral sclerosis (PLS) bears close resemblance to cases of amyotrophic lateral sclerosis (ALS) presenting with spasticity, but histopathological studies have shown significant difference between the two conditions. When the lower motor neurons in cases of ALS and PLS are compared with the equivalent cells of control subjects, morphometric...

Eight patients with a homogeneous syndrome of progressive symmetric spinobulbar spasticity were studied. Clinical features were limited to those associated with dysfunction of the descending motor tracts and included spastic quadriparesis, pseudobulbar affect, spastic dysarthria, hyper-reflexia and bilateral Babinski signs. Lower motor neuron findi...

We have examined the gastric luminal content of Na+, K+, and protein and mucosal levels of myeloperoxidase in rats between the ages of 10 and 60 days in response to luminal instillation of ethanol (20 and 50% w/v). In control animals the appearances of ions and protein and myeloperoxidase activities were low and similar in all age groups. Luminal c...

It has been suggested that the degeneration of lower motor neurons in amyotrophic lateral sclerosis (ALS) is a transneuronal event, secondary to the loss of corticospinal and corticobulbar nuerons, In an attempt to test this hypothesis, the cross-sectional areas of phyramidal cells in layer 5 of the foot and tongue areas of the precentral gyri were...

7b the Editor.— The article "Frozen Pre-embryos"1 by the Board of Trustees of the American Medical Association contains the following statement: "Because fertilized eggs are frozen before the embryonic stage of development, they are referred to as pre-embryos."The early stages of development, including the formation of the blastocyst and inner cel...

Previous studies of the permeation of circulating protein tracers yielded conflicting results in the enteric nervous system. We show that within 5 min of intravenous injection, horseradish peroxidase (HRP) enters all the extracellular spaces of the ganglia and connecting strands of the submucous and myenteric plexuses of the rat's small intestine,...

Immunohistochemical methods were used to determine the localisation of immunoreactivities to a variety of antigens involved in neurotransmission in the myenteric plexus of the colon in the rat and mouse. The findings in the two species were closely similar. Five neuronal types have been identified. (i) The axons of extrinsic noradrenergic sympathet...

Earlier reports have suggested that anthraquinone purgatives in excessive amounts cause degeneration of neurons in the enteric nervous system. Danthron (1,8-dihydroxyanthraquinone) was administered to rats in their drinking water for four months. The effects of the drug on the total number of neurons and on the immunoreactivity of eight putative ne...

In the trachea and bronchi of the atropinized rat, the proportion of degranulating mast cells (defined as having one or more granules outside the body of the cell in a 10-microns thick section) was increased from 35-40% to 48-55% following electrical stimulation of one or both vagus nerves for 3 min. The increase occurred bilaterally, though it was...

Removal of the salivary glands (SALX) in rats has been shown to increase the susceptibility of gastric mucosa to ulcerogens. In the present study, we have investigated the role of specific salivary glands in this response. In addition, we have examined whether a functional link exists between the salivary glands, epidermal growth factor (EGF), and...

Effects of senna on the myenteric plexus of the colon were investigated in view of earlier reports that this anthraquinone cathartic depletes the plexus of its intrinsic neurons. Rats and mice were given purgative doses of sennosides in their drinking water for 4 and 5 months, respectively. Body growth was reduced, and the weight of the colon with...

A stenosis is produced when a rat's transected small intestine is repaired with a conventional inverting line of silk or catgut sutures. In the new technique, the cut surfaces are apposed over a splint made of dry macaroni of suitable diameter, and then joined end-to-end with polyglactin stitches. The internal splint is quickly softened and digeste...

1. We have investigated the binding of the tritiated forms of prostaglandin E2 (PGE2) and a stable analogue of prostacyclin (Iloprost) to isolated cells of rabbit oxyntic mucosa. 2. The highest degree of specific [3H]PGE2 binding occurred in a cellular fraction enriched in parietal cells. [3H]Iloprost binding occurred predominantly in cells identif...

Wholemounts consisting of both muscular layers of the rat's intestine have been stained with Cuprolinic blue and magnesium chloride. The procedure, which is specific for RNA, gives selective coloration of the somata of the neurons of the myenteric plexus. The neuronal nucleoli and Nissl substance are the only blue structures in such preparations. F...

Reinnervation of the skeletal muscle in the tongue following vago-hypoglossal anastomosis was studied by means of retrograde labelling with horseradish peroxidase and anterograde labelling with the autoradiographic tracing method combined with acetylcholinesterase staining for motor endplates. The proximal stump of the transected vagus nerve was an...

Nature is the international weekly journal of science: a magazine style journal that publishes full-length research papers in all disciplines of science, as well as News and Views, reviews, news, features, commentaries, web focuses and more, covering all branches of science and how science impacts upon all aspects of society and life.

Chromic salts have been studied as fixatives of mammalian tissues for light microscopy, and the binding of the metal has been examined histochemically. Tissues bind chromium(III) from aqueous solutions less acid than pH 2.5; the metal attaches mainly to collagen and basement membranes. Solutions containing chromium(III) as the only active ingredien...

The optic nerve of the rat has been examined by light and electron microscopy, and also for vascular permeability to fluorescently labelled albumin, 2 days to 34 weeks after crushing in the orbit. The operation was usually followed by loss of 20-70% of the retinal ganglion cells. Axons could be followed from the retina into the optic nerve at all p...

Atrophy in a denervated muscle results from the disuse caused by paralysis of the muscle, and from the loss of special nerve-derived trophic substances. Crude preparations of protein from rat or sheep sciatic nerves have been shown to prevent the nondisuse atrophy of the rat's extensor digitorum longus muscle when injected into the denervated muscl...