Fig.3 AgNOR staining with peripheral blood lymphocytes. . The technical aspect of AgNOR staining procedure should be carefully dealt with. Attention should. Conclusions: AgNOR staining can be considered as a useful adjunct to adjunct procedures which can diagnose malignancy at the earliest. Silver staining results are comparable to those obtained. [A standardized AgNOR stain method for formalin fixed and paraffin embedded .. to be a good adjunct to presently available frozen section techniques and cytology.

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Although amount of silver nitrate is in some relation to the amount of activity, or number of active, rDNA copies; it cannot be used for an accurate quantitative study, partly because of differences of silver staining strengths within and between slides depending on fixation procedures, density of nuclei, preparation quality and age of slides.

Modified method of AgNOR staining for tissue and interpretation in histopathology

Improvement in the staining and the visualization of the argyrophilic proteins of the nucleolar organizer region at the optical level. For quick results incubate at C for min. If black precipitate occurs, slides were not clean or the silver nitrate solution was contaminated. In previous methods counter stain Neutral red was used.

Modified method of AgNOR staining for tissue and interpretation in histopathology

Small-capped bottles were used in which different quantities of silver nitrate like 1, 2, 3 or 4 g were kept. Two grams of silver nitrate and hence 6 ml of final working solution were found sufficient to stain five sections.

AgNOR analysis in multiple myeloma allowed the identification of atypical plasma cells with different proliferative activity in bone marrow biopsies Pich et al.

We compared the results both with and with out counter stain with both tonic agents gold nitrate, sodium thiosulphate Table 1. Courses and Science Pictures. Rinse in distilled water. The grading of size variation was performed according to Khan et al. Too long staining results in black nucleoli and eventually black nuclei. These results are consistent with the findings of Khan et al. Support Center Support Center. Solution B It comprised silver nitrate and deionized water.


Should not be used because of the blurring of AgNOR staining. The mean dispersion of AgNOR per cell in normal tissue and reactive gliosis was nil, while low in low-grade astrocytoma and high in high-grade astrocytoma mean 1. It is very costly and use the amount required for the number of cases. The quantity of silver nitrate adequate to stain the number of slide in a particular batch was used. The tissue was then incubated in acid alcohol three parts ethanol: After silver-staining, the NORs can be easily identified as black dots exclusively localised throughout the nucleolar area, and are called “AgNORs”.

Four-micron unstained sections were obtained from the selected areas of tissue to perform the silver staining.

Silver Staining for Nucleolar Organizer Regions

Over the past 12 years, the “AgNOR method” has been applied in tumour pathology for both diagnostic and prognostic purposes. Nuclei should be visible without phase contrast.

Silver nitrate stains clothes lab benches and fingers black, normally after about 6h. We selected 60 samples, 10 samples each of normal brain tissue, reactive gliosis, pilocytic astrocytoma, astrocytoma grade II, astrocytoma grade III, and glioblastoma multiform grade IV astrocytoma. Our results are in accordance with studies on bladder carcinoma and multiple myeloma showing association between mean AgNOR count and histological grade of differentiation Pich et al.

We recommend the use of morphometry for AgNOR size in future. In our study, the results of size and dispersion of AgNORs were clearer without the use of counterstain. It was helpful in the classification of urothelial carcinoma in different grades of malignancy and in subdividing patients with grade II transitional cell carcinomas into low- and high-risk groups Helpap et al.


Estimation of AgNORs parameters number, size and distribution has been applied in tumour pathology both for diagnostic and prognostic purposes.

Make sure nylon mesh is soaked thoroughly. There was moderate to high variation in the size of AgNOR dots in high procedurre astrocytoma mean 1.

Nucleolus organizing region activity. For storage, leave the slide on the bench until all the xylene has evaporated, remove the coverslip carefully and put slide in a box.

AgNOR staining and quantification.

This method gives excellent contrast, but you have to cope with a little bit of xylene smell. In our study, the mAgNOR and pAgNOR proliferative index were high in more proliferative and mitotically active tumours cells as we moved up in different grades of astrocytoma i.

AgNOR size and dispersion may be more important than counting alone: Naveed, who provided the opportunities and facilities for this research work and encouraged to write it for international readers. One of these was originally introduced by Ploton et al. Patients and methods We selected 60 samples, 10 samples each of normal brain tissue, reactive gliosis, pilocytic astrocytoma, astrocytoma grade II, astrocytoma grade III, and glioblastoma multiform grade IV astrocytoma.

Relationship between AgNOR proteins, Ki antigen, p53 immunophenotype and differentiation markers in archival breast carcinomas. Abstract This study was conducted in the department of Pathology King Edward Medical University, from June to December to introduce the new method of AgNOR staining and its interpretation to increase its reliability.