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Acid ethanol fixation and polyester wax embedding combines preservation of antigenic determinants with good
P J Roholl1, H F Dullens, J Kleijne
1IVEG-TNO, Rijswijk, The Netherlands.
Insights
This study presents a new method for preserving lymphoid tissues and blood cells using polyester wax. This technique enhances cell visibility for accurate immunophenotyping and cell identification in research.
Area of Science:
- Immunology
- Cell Biology
- Histology
Background:
- Accurate immunophenotyping of lymphoid tissues and blood cells is crucial for various research applications.
- Traditional tissue preservation methods can sometimes compromise cellular morphology and antigenicity.
Purpose of the Study:
- To evaluate the efficacy of acid ethanol fixation and polyester wax embedding for preserving lymphoid tissue and peripheral blood mononuclear cells.
- To assess the suitability of this method for detailed cytomorphological analysis and immunophenotyping.
Main Methods:
- Lymphoid tissues and peripheral blood mononuclear cells were fixed in acid ethanol.
- Samples were embedded in polyester wax with a melting point of 37°C.
- Sections were stained with various poly- and monoclonal antibodies for immunophenotyping.
Main Results:
- The polyester wax embedding method preserved excellent cytomorphology, enabling differentiation of various lymphoid and non-lymphoid cells.
- Satisfactory immunophenotyping was achieved for histiocytes, endothelial, mesenchymal, epithelial cells, lymphocytes (including subtypes), and lymphokine-activated killer (LAK) cells.
- Staining patterns on polyester wax sections were comparable to those on frozen sections, with successful double labeling of bromodeoxyuridine-incorporated cells.
Conclusions:
- Acid ethanol fixation and polyester wax embedding is a valuable technique for preserving cellular detail and antigen integrity in lymphoid tissues and blood cells.
- This method facilitates accurate immunophenotyping and cytomorphological analysis, comparable to frozen sections.
- The technique supports advanced applications like double labeling for cell proliferation studies.
Abstract:
Lymphoid tissue, and/or isolated peripheral mononuclear blood cells were fixed in acid ethanol and embedded in polyester wax (melting point 37 C). The excellent cytomorphology obtained allowed distinguishing different types of individual lymphoid and nonlymphoid cells. Furthermore, this procedure was satisfactory in the immunophenotyping of histiocytes, endothelial, mesenchymal, epithelial cells, different (sub-) types of lymphocytes and also of lymphokine activated killer (LAK) cells. The staining patterns obtained with the different poly- and monoclonal antibodies on polyester wax sections were not only analogous to those obtained on frozen sections, but cells which had incorporated bromodeoxyuridine could be double labeled with specific antiserum.
