Combined Diaminobenzidine–Colloidal Gold Staining

Mutasa, H. C. F.

doi:10.1016/b978-0-12-333929-4.50019-9

Cited by 3 publications

(1 citation statement)

References 46 publications

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“…Labelling density ( LD ) expresses the number of observed gold particles per length or area unit (Bendayan et al , 1980; Bendayan, 1984b; Posthuma et al , 1987; Kalina et al , 1988; Mutasa, 1991; Bogers et al , 1996). This method is easily performed by counting all gold particles falling within the cellular compartment by applying at random a bi‐dimensional lattice over micrographs (Gundersen, 1977; Lucocq, 1994), called the counting frame.…”

Section: Labelling Estimators and Stereological Analysismentioning

confidence: 99%

Quantifying immunogold labelling in transmission electron microscopy

D'Amico¹,

Skarmoutsou

2008

Journal of Microscopy

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Summary Gold particles labelling on ultrathin sections is extensively used for antigen localization in transmission electron microscopy. In establishing absolute or relative counts in tissue sections, it would be expedient to use stereologically based unbiased estimates for quantitative results. Nowadays, quantitative immunoelectron microscopy has achieved good and satisfactory results to test whether the gold labelling follows a non‐random or a random pattern and then to draw statistical comparisons between cell subcompartments within a sample of cells or between experimental groups of cells. This brief informal review of literature focuses on the relative quantitative determinations of gold labelling of antigens as well as on the statistical distribution comparisons in transmission electron microscopy.

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Section: Labelling Estimators and Stereological Analysismentioning

confidence: 99%

Quantifying immunogold labelling in transmission electron microscopy

D'Amico¹,

Skarmoutsou

2008

Journal of Microscopy

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Osmium Tetroxide-Potassium Ferrocyanide Intensification of a Diaminobenzidine Product Obtained by Photoconversion of a Fluorescent Label: a Study of Human Neutrophil Granules

Mutasa

1995

Biotechnic & Histochemistry

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A diaminobenzidine posttreatment employing osmium tetroxide and potassium ferrocyanide was successfully used to intensify the diaminobenzidine stain formed by photoconversion of immunofluorescent labelling. Lactoferrin labelled granules became visible following the photoconversion process. Adequate diaminobenzidine staining was obtained after irradiating the cytospin preparations with ultraviolet light for 30-40 min. The diaminobenzidine stain had advantages over the fluorescent stain owing to its greater stability, greater density, and ability to be intensified. The enhancement procedure intensified both the color and density of the diaminobenzidine product. Consequently, shorter irradiation times could be used. Osmium tetroxide solutions of 3-4% increased the intensity with minimal background staining. Ultrastructural immunogold cytochemistry on ultrathin sections confirmed the existence of the lactoferrin labelled structures observed by light microscopy of cytospin preparations indicating that these were secondary granules. The photomicroscopy procedures used in this study were simple to perform and could be applied to studies of other cellular antigens prior to using immunoultramicroscopy.

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Changes in Neutrophil Granule Protein and Cytoplasmic Fibrils in Human Acute Myeloid Leukemias

Mutasa

1995

Biotechnic & Histochemistry

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Granule protein deficiencies in morphologically mature neutrophil cells of peripheral blood from human patients with acute myeloid leukemia was demonstrated using post-embedding immunocytochemistry. Abnormal immunoreactivity of granule proteins was detected in seven of nine patients. Decreased immunoreactivity patterns were found more for the primary granule markers elastase and myeloperoxidase than for the secondary granule marker lactoferrin. Leukemias with a predominant myeloid component, in contrast to those with a predominant monocytoid component, had more neutrophil cells showing immunodeficiencies for one or more granule markers. The proportion of neutrophil cells showing immunodeficiencies varied greatly for each granule marker; more variation was obtained for elastase, lactoferrin and myeloperoxidase than for lysozyme, possibly because lysozyme is a marker for both granule types. In addition, no correlation could be found between any of the immunoreactivity deficiencies for the neutrophil granule glycoproteins elastase, lactoferrin, lysozyme and myeloperoxidase and the abundance of a particular set of ultrastructural features in the circulating leukemic cells from any of the nine patients. Nonetheless, most of the immature myeloid cells from peripheral blood of leukemic patients showing neutrophil protein immunoreactivity abnormalities in one or more granule markers often and randomly displayed one or more unusual ultrastructural features. The clinical and pathological significance of neutrophil granule protein deficiencies and the abundance of fibrillar structures in malignant myeloid cells presently is uncertain.

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Combined Diaminobenzidine–Colloidal Gold Staining

Cited by 3 publications

References 46 publications

Quantifying immunogold labelling in transmission electron microscopy

Quantifying immunogold labelling in transmission electron microscopy

Osmium Tetroxide-Potassium Ferrocyanide Intensification of a Diaminobenzidine Product Obtained by Photoconversion of a Fluorescent Label: a Study of Human Neutrophil Granules

Changes in Neutrophil Granule Protein and Cytoplasmic Fibrils in Human Acute Myeloid Leukemias

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