Histochemical and immunohistochemical techniques on acrylate embedded bone biopsies

Vykoupil, K. F.; Thiele, Jürgen; Georgii, A.

doi:10.1007/bf00995915

Cited by 28 publications

(4 citation statements)

References 9 publications

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“…Tissues fixed and embedded at 4°C in methacrylate (Ashford et al, 1972;Vykoupil et al, 1976;Beckstead & Bainton, 1980;Beckstead et al, 1981;Dobyan et al, 1982;Beckstead, 1983;Namba et al, 1983;Soufleris et al, 1983;Litwin, 1985;Bogdanffy et al, 1986;Murray et al, 1987;Pretlow et al, 1987a,b;Turner et al, 1987) have been used for an increasing number of studies including the analyses of stromal cells in human colonic carcinoma (Pretlow et al, 1984;Luebbers et al, 1985). This study with 24~m-thick sections of methacrylate-embedded segments of normal rat colon facilitated detection and/or more precise localization of enzyme activities (Fig.…”

Section: Discussionmentioning

confidence: 99%

In situ localization of enzymes and mucin in normal rat colon embedded in plastic

et al. 1989

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Several enzymes were investigated histochemically in the colons of normal male F344 rats in order to understand the function of different types of cells in this tissue. Serial methacrylate-embedded sections (2-4 microns) allowed the precise localizations of several enzymes including acid phosphatase, alkaline phosphatase, gamma-glutamyl transpeptidase, N-acetyl-beta-D-glucosaminidase (hexosaminidase), alpha-naphthyl butyrate esterase and 5'-nucleotidase. Sites reactive with periodic acid-Schiff were also localized. Gradients of enzyme activity were observed between caecum and rectum and/or from the luminal surfaces to the bases of the crypts for hexosaminidase, esterase and gamma-glutamyl transpeptidase. To our knowledge this is the first histochemical demonstration of gamma-glutamyl transpeptidase in normal rat colonic epithelial cells. The utilization of the methacrylate-embedding technique has revealed previously undescribed gradients of enzyme activity and has allowed the localization of enzyme activities not previously reported in normal rat colonic mucosa.

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Section: Discussionmentioning

confidence: 99%

In situ localization of enzymes and mucin in normal rat colon embedded in plastic

et al. 1989

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“…In addition to the smears of peripheral blood, trephine biopsies of the bone marrow were performed following the technique of Burkhardt (19)(20)(21) or Jamshidi & Swaim (22). Further processing included resin embedding, semithin sectioning and several staining procedures (Giemsa, Gomori's silver impregnation, Goldner's trichrome, methylgreen-pyronine, Prussian Blue reaction and periodic acid-Schiff (PAS) as recorded by Vykoupil et al (23). From marrow samples of the 6th patient (Table 1) the following cytochemical and immunohistochemical reactions were applied on smears and sections: myelo-and platelet peroxidase, naphthol-AS-D-chloroacetate esterase, Sudan-Black B, alkaline phosphatase and antifactor VIII.…”

Section: Examination Of the Bone Marrowmentioning

confidence: 99%

Malignant (acute) myelosclerosis ‐ a clinical and pathological study in 6 patients

Thiele

Krech

Vykoupil

et al. 1984

Scandinavian Journal of Haematology

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Clinical and morphological findings are described in 6 patients with malignant (acute) myelosclerosis/fibrosis (MMS). Haematological data are characterized by severe anaemia and thrombocytopenia, frequently accompanied by a leucopenia with an increase in myeloblasts and promyelocytes in the peripheral blood count. There is an absence of, or a minimal hepatosplenomegaly and the survival times after onset of clinical symptoms to death range from 4–12 months. The histopathology of the bone marrow shows a conspicuous proliferation of blasts (myeloblasts, promyelocytes and megakaryoblasts) in a variable amount, besides a fibrosclerosis consisting of reticulin and collagen fibrils. A comparison of MMS with ordinary myelofibrosis/osteomyelosclerosis (MF/OMS) of a chronic course implicates two important facts: (a) evolution of fibrosclerosis takes a considerable period of time for manifestation, which ranges between 20–30 months; (b) the histopathology of MMS is identical with those features observable in the rare event of a terminal stage, i.e. blastic transformation of chronic MF/OMS. Consequently MMS should be designated as an accelerated variant of MF/OMS with a rather early occurrence of a blastic crisis. The insidious onset with the dominant clinical finding of anaemia is probably responsible for the relatively late appearance of symptoms, while the progressive course prevents an overt myeloid metaplasia with a massive hepatosplenomegaly.

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“…In 1964, Farnes and Barker (6) applied the enzyme stain to the analysis of the composition of different cell types in bone marrow. Although the preservation of acid phosphatase staining has been studied in regard to fixation and processing (7)(8)(9)(10)(11)(12), embedding material (13)(14)(15)(16)(17)(18)(19), and substrates used (20)(21)(22)(23)(24)(25)(26), there has not been a study to determine the storage conditions of blocks and unstained slides for optimal preservation of the enzyme. Accurate representation of alkaline and acid phosphatase enzymes in bone requires attention to more than the fixation, processing, and staining protocols.…”

Section: Introductionmentioning

confidence: 99%

The Effects of Storage Conditions on the Preservation of Enzymatic Activity in Bone

Cosby¹,

Troiano²,

Kacena³

2008

journal of histotechnology

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Alkaline phosphatase and acid phosphatase are two major enzymatic measures of osteoblastic and osteoclastic activity, respectively. As a result, the preservation of the enzymes in bone specimens to near in vivo accuracy is essential. Despite standardization of the staining process, several factors related to the storage of blocks and slides before sectioning and staining impact the level of enzymes detected in the tissue. Block condition (intact, faced, or unstained) as well as environment (temperature and length of time in storage) affect alkaline phosphatase preservation while the acid phosphatase enzyme remains unaffected. We conclude that to optimally preserve alkaline phosphatase enzyme, methacrylate-embedded undecalcified murine bones should be stored as intact blocks. After sectioning, the faced blocks should be stored at 4°C for optimal enzyme staining of future sections. Furthermore, it is best to stain sections immediately after sectioning.

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Histochemical and immunohistochemical techniques on acrylate embedded bone biopsies

Cited by 28 publications

References 9 publications

In situ localization of enzymes and mucin in normal rat colon embedded in plastic

In situ localization of enzymes and mucin in normal rat colon embedded in plastic

Malignant (acute) myelosclerosis ‐ a clinical and pathological study in 6 patients

The Effects of Storage Conditions on the Preservation of Enzymatic Activity in Bone

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