Novel polychrome staining distinguishing osteochondral tissue and bone cells in decalcified paraffin sections

The morphologic pathologist may require information on the ultrastructure of a non-specific lesion seen under the light microscope before he can make a specific determination. Such lesions, when caused by infectious disease agents, may be sparsely distributed in any organ system. Tissue culture systems, too, may only have widely dispersed foci suitable for ultrastructural study. In these situations, when only a few, small foci in large tissue areas are useful for electron microscopy, it is advantageous to employ a methodology which rapidly selects a single tissue focus that is expected to yield beneficial ultrastructural data from amongst the surrounding tissue. This is in essence what "LIFTING" accomplishes. We have developed LIFTING to a high degree of accuracy and repeatability utilizing the Microlift (Fig 1), and have successfully applied it to tissue culture monolayers, histologic paraffin sections, and tissue blocks with large surface areas that had been initially fixed for either light or electron microscopy.

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Pathology of Bone Cells in Pigs with Experimental Turbinate Osteoporosis (Atrophic Rhinitis)

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100065560 ◽

1971 ◽

Vol 29 ◽

pp. 304-305

Author(s):

A. W. Fetter ◽

C. C. Capen

Keyword(s):

Mucous Membrane ◽

Bone Cells ◽

Atrophic Rhinitis ◽

Infectious Agents ◽

Metabolic Disturbances ◽

Progressive Reduction ◽

Naturally Occurring ◽

Fundamental Cause ◽

Osteocytic Osteolysis ◽

Uncertain Etiology

Atrophic rhinitis in swine is a disease of uncertain etiology in which infectious agents, hereditary predisposition, and metabolic disturbances have been reported to be of primary etiologic importance. It shares many similarities, both clinically and pathologically, with ozena in man. The disease is characterized by deformity and reduction in volume of the nasal turbinates. The fundamental cause for the localized lesion of bone in the nasal turbinates has not been established. Reduced osteogenesis, increased resorption related to inflammation of the nasal mucous membrane, and excessive resorption due to osteocytic osteolysis stimulated by hyperparathyroidism have been suggested as possible pathogenetic mechanisms.The objectives of this investigation were to evaluate ultrastructurally bone cells in the nasal turbinates of pigs with experimentally induced atrophic rhinitis, and to compare these findings to those in control pigs of the same age and pigs with the naturally occurring disease, in order to define the fundamental lesion responsible for the progressive reduction in volume of the osseous core.

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In situ microprobe quantitation of inorganic particle burden in paraffin sections of lung tissue

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100107010 ◽

1988 ◽

Vol 46 ◽

pp. 990-991

Author(s):

Jerrold L. Abraham

Keyword(s):

Lung Tissue ◽

Quantitative Information ◽

Particulate Material ◽

Paraffin Sections ◽

Tissue Samples ◽

Qualitative And Quantitative ◽

The Past ◽

Quantitative Analyses ◽

Data Result

Inorganic particulate material of diverse types is present in the ambient and occupational environment, and exposure to such materials is a well recognized cause of some lung disease. To investigate the interaction of inhaled inorganic particulates with the lung it is necessary to obtain quantitative information on the particulate burden of lung tissue in a wide variety of situations. The vast majority of diagnostic and experimental tissue samples (biopsies and autopsies) are fixed with formaldehyde solutions, dehydrated with organic solvents and embedded in paraffin wax. Over the past 16 years, I have attempted to obtain maximal analytical use of such tissue with minimal preparative steps. Unique diagnostic and research data result from both qualitative and quantitative analyses of sections. Most of the data has been related to inhaled inorganic particulates in lungs, but the basic methods are applicable to any tissues. The preparations are primarily designed for SEM use, but they are stable for storage and transport to other laboratories and several other instruments (e.g., for SIMS techniques).

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Discovery of intracellular 2,8 dihydroxyadenine crystals in bovine lymphatic and hepatic cells

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s042482010012881x ◽

1987 ◽

Vol 45 ◽

pp. 906-907

Author(s):

W. E. Rigsby ◽

D. M. Hinton ◽

V. J. Hurst ◽

P. C. McCaskey

Keyword(s):

Polarized Light ◽

Paraffin Sections ◽

Tissue Samples ◽

Whole Cells ◽

Tissue Sections ◽

Hepatic Cells ◽

Slaughter House ◽

Mammalian Tissues ◽

Carbon Coated ◽

Cellular Components

Crystalline intracellular inclusions are rarely seen in mammalian tissues and are often difficult to positively identify. Lymph node and liver tissue samples were obtained from two cows which had been rejected at the slaughter house due to the abnormal appearance of these organs in the animals. The samples were fixed in formaldehyde and some of the fixed material was embedded in paraffin. Examination of the paraffin sections with polarized light microscopy revealed the presence of numerous crystals in both hepatic and lymph tissue sections. Tissue sections were then deparaffinized in xylene, mounted, carbon coated, and examined in a Phillips 505T SEM equipped with a Tracor Northern X-ray Energy Dispersive Spectroscopy (EDS) system. Crystals were obscured by cellular components and membranes so that EDS spectra were only obtainable from whole cells. Tissue samples which had been fixed but not paraffin-embedded were dehydrated, embedded in Spurrs plastic, and sectioned.

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Detection of atrial natriuretic peptides (ANP) in rat atria by immunogold-silver staining (IGSS)

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100169250 ◽

1994 ◽

Vol 52 ◽

pp. 304-305

Author(s):

Robyn Rufner ◽

Gerhard W. Hacker ◽

Michele Forte ◽

Nancyleigh E. Carson ◽

Cristina Xenachis ◽

...

Keyword(s):

Natriuretic Peptides ◽

Silver Staining ◽

Sodium Thiosulfate ◽

Atrial Natriuretic Peptides ◽

Paraffin Sections ◽

Glass Slides ◽

Metallic Salts ◽

Wistar Kyoto ◽

Right Atria

The use of immunogold-silver staining (IGSS) to enhance label penetration and Localization for immunocytochemistry or in situ hybridization utilizing a variety of metallic salts has been documented. In this morphological study, the effects of silver acetate, silver lactate and silver nitrate were evaluated for immunogold-labeling of a trial natriuretic peptides (ANP) in rat right atria.Eight Wistar Kyoto retired breeders were sedated with pentobarbital, perfused with either 4% paraformaldehyde (LM) or Karnovsky's fixative (EM), and right atria were dissected, processed, embedded in paraffin or epon, respectively and sectioned according to conventional methods. For light microscopy, an indirect IGSS method according to Hacker (3) was performed. Paraffin sections on glass slides were washed in ddH2O, immersed in Lugol's iodine, washed in ddH2O and treated with 2.5% aqueous sodium thiosulfate for 20 sec. After additional washes in ddH2O and TBS-0.1% fish gelatin, 10% normal goat serum (PBS with 1% BSA) was applied for 20 min before an overnight incubation at 4°C with a polyclonal α-ANP primary antibody (Peninsula Labs, 1:1000 in TBS/BSA).

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In vitro - studies with antler bone cells: Structure forming capacity, osteocalcin production and influence of sex steroids

Osteologie/Osteology ◽

10.1024/1019-1291.15.4.245 ◽

2006 ◽

Vol 15 (04) ◽

pp. 245-257 ◽

Cited By ~ 3

Author(s):

H. J. Rolf ◽

K. G. Wiese ◽

H. Siggelkow ◽

H. Schliephake ◽

G. A. Bubernik

Keyword(s):

Sex Steroids ◽

Bone Cells ◽

In Vitro Studies ◽

Osteocalcin Production

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Platelet Distribution and Survival in Plasma Clots and Thrombi

Thrombosis and Haemostasis ◽

10.1055/s-0038-1654578 ◽

1961 ◽

Vol 06 (03) ◽

pp. 470-484 ◽

Cited By ~ 2

Author(s):

Peter Wolf

Keyword(s):

Blood Vessels ◽

Calcium Ions ◽

Paraffin Sections ◽

Cardiac Infarction ◽

Cardiac Blood ◽

Before And After

SummaryViscous metamorphosis of platelets in native and sequestrene plasma, before and after thrombin and plasmin action, has been studied. A method for the examination of platelet distribution in plasma clots and tissue paraffin sections is described.It was found that not all platelets undergo viscous metamorphosis in plasma clots or in intravascular thrombi. Platelets before and after viscous metamorphosis are not digested by plasmin. After plasmin action intact platelets can still undergo viscous metamorphosis and the fibrils which are then produced are not made of fibrin.Thrombin in the absence of calcium ions will not cause platelets to undergo viscous metamorphosis.Total blockage of cardiac blood vessels by platelet masses in cases of cardiac infarction is demonstrated. The significance of these findings in relation to blood sludging, and future lines of treatment are discussed.

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