Strategies to maximize adhesion of thick paraffin sections of the brown trout liver for stereological purposes

• Type: Article in International Scientific Journal
• Date: 2001

Authors

marcos, r (Author) Other; rocha, e (Author) ICBAS; monteiro, raf (Author) ICBAS

Journal

Title: Journal of Histotechnology

Vol. 24

Pages: 37-42

ISSN: 0147-8885

Publisher: Taylor & Francis

Indexing

ISI Web of Knowledge - 1 Citation

Scopus - 3 Citations

Scientific classification

FOS: Natural sciences > Biological sciences

Other information

Authenticus ID: P-000-W98

Abstract (EN): Thick histological sections are essential for implementing many modern stereological techniques because most rely on the optical disector technique both for sampling and counting objects (namely cells). In conducting any stereological evaluation, objects must be unambiguously identified. In many instances, only immunocytochemistry allows the required clear cell identification in sections. In immunohistochemical procedures adhesion media are routinely used, especially when including aggressive treatments for antigen retrieval leg, microwave exposure) and tissue permeabilization leg, Triton X-100). Immunohistochemical techniques performed on thick sections (30-40 mum) can cause additional adhesion problems, reinforcing the need to seek the best adhesion strategies for particular situations. Formalin fixed liver of the brown trout was used in this study. After pre-treating the slides with 2N hydrochloric acid (HCl) and acetone, we compared 4 of the most used adhesion media, namely Mayer solution, poly-l-lysine, aminopropyltriethoxysilane (APES), and chromalum-gelatin. A control procedure with a pre-treatment step was also used. For each adhesion medium and the control, 5 thick paraffin sections (40 mum) were used. After aggressive treatment with microwave irradiation and Triton X-100 and with an immunohistochemical avidin-biotin-like protocol, the sections were blindly and semiquantitatively evaluated. Only in treatments with APES and chromalum-gelatin was the adhesion satisfactory. APES was slightly more effective. A short illustration exercise using the single hepatocyte nucleus as the counting unit demonstrated the feasibility of the optical disector. This technique offers the best option because the chromalum-gelatin costs less and there are only small differences between the 2 strategies.

Language: English

Type (Professor's evaluation): Scientific

No. of pages: 6