Related Experiment Video
Updated: Jun 17, 2026
Working with Human Tissues for Translational Cancer Research
Published on: November 26, 2015
Processing of tissue specimens
1Department of Gynecologic and Breast Pathology, Armed Forces Institute of Pathology, Washington, DC, USA.
Insights
Tissue specimens require fixation, paraffin embedding, and thin sectioning for antibody testing. Pretreatment steps like rehydration, antigen retrieval, and blocking are crucial for accurate results.
Area of Science:
- Histopathology
- Immunohistochemistry
Background:
- Antibody testing of tissue specimens is essential for diagnosis.
- Proper tissue preparation is critical for successful antibody detection.
Purpose of the Study:
- To outline the standard procedure for preparing tissue specimens for antibody testing.
- To detail the critical steps involved in tissue fixation, embedding, sectioning, and pretreatment.
Main Methods:
- Tissue fixation using 10% buffered formalin.
- Paraffin embedding and microtome sectioning.
- Rehydration, antigen retrieval (enzymatic or heat-based), and blocking of endogenous peroxidase.
Main Results:
- Standardized protocols ensure tissue integrity and antigen accessibility.
- Effective pretreatment optimizes antibody binding and signal detection.
- Properly prepared slides are ready for antibody incubation.
Conclusions:
- Successful antibody testing relies on meticulous tissue preparation and pretreatment.
- Adherence to established protocols is key for reproducible and accurate histopathological analysis.
Abstract:
In order to test tissue specimens with antibody, they first have to be preserved in fixative, embedded in paraffin, and sectioned very thinly onto glass microscope slides. Any piece of tissue, immediately after excision, must be placed into an adequate volume of fixative. Fixatives vary, but the standard one is 10% buffered formalin. After an optimum fixation time (for formalin, about 16 h), the sample must be embedded in paraffin and sectioned on a microtome. Paraffin-embedded sections placed on positively charged slides (either coated or commercially prepared) are then ready for various pretreatment steps. First, the paraffin must be replaced with water through a series of rehydration steps. Then, depending on the antigen to be tested, the section can be proteolytically digested with enzymes or heat-treated in low or high pH solutions. Following that, the endogenous peroxidase enzyme or oxidative compounds can be quenched in a hydrogen peroxide solution. The sections are then ready to be tested with antibody after an incubation in a normal serum solution blocks any available charged sites.
