Related Experiment Video
Updated: Apr 24, 2026
Histological-Based Stainings Using Free-Floating Tissue Sections
Published on: August 25, 2020
Destaining hematoxylin and eosin stains and restaining for immunohistochemistry has diagnostic value for cytology
Mohammad M Al-Attar1, Tianle Zou2, Salwa Khedr2
1Department of Pathology, Mass General Brigham, Boston, United States.
Insights
Destaining and restaining (DSRS) improves immunohistochemical (IHC) interpretation accuracy for sparse neoplastic cells. This technique enhances diagnostic accuracy in challenging cytology cases, offering a cost-effective solution.
Area of Science:
- Pathology
- Cytopathology
- Immunohistochemistry
Background:
- Immunohistochemical (IHC) staining interpretation is challenging with sparse, mixed, or obscured neoplastic cells.
- Accurate IHC requires precise localization of antibodies to specific neoplastic cells.
Purpose of the Study:
- To develop and validate a destaining and restaining (DSRS) technique for IHC on cytology cell block sections.
- To assess the utility of DSRS for improving diagnostic accuracy in limited fine needle aspiration (FNA) samples.
Main Methods:
- A destaining and restaining (DSRS) protocol was applied to hematoxylin and eosin (H&E) stained cytology cell block sections.
- Slide scanning and image synchronization were used to track neoplastic cells after DSRS and IHC.
- IHC results were validated without protocol modification.
- Diagnoses from limited FNA samples using DSRS were compared to those from more ample specimens.
Main Results:
- The DSRS technique did not compromise tissue or IHC quality.
- Diagnostic concordance between limited FNA samples (using DSRS) and final diagnoses improved to 89% (8/9) from 22% (2/9) prior to DSRS.
- Immunostain interpretation accuracy was 100% (9/9) with the DSRS method.
Conclusions:
- DSRS enables accurate IHC staining attribution to specific cells in challenging cytology specimens.
- This technique is valuable for diagnosing cases with sparse cellularity, mixed cell populations, or obscured cell morphology.
- DSRS offers an economical alternative to advanced techniques like multiplex IHC or immunofluorescence.
Objectives:
The use of one antibody per slide for immunohistochemical (IHC) studies is difficult to interpret when neoplastic cells are sparse, mixed with complex mixtures of other cells, or are obscured by the IHC stain itself. To accurately assign IHC results to particular neoplastic cells, we developed and validated a technique of destaining hematoxylin and eosin (H&E) stains and restaining (DSRS) by IHC on cytology cell block sections and studied its utility.
Materials And Methods:
We identified 9 patients with fine needle aspiration (FNA) samples performed for a variety of tumors. Specimens were collected and made into ThinPreps and cell blocks. Cell block serial H&E stains revealed a rich background of benign cells, with rare scattered atypical cells arranged singly or in small clusters. Select H&E slides were scanned, destained, and then restained with one IHC biomarker per H&E. Slide scanning and image synchronization were used in tracking neoplastic cells. IHC results were validated for the DSRS process for all antibodies without the need for modification of the IHC protocol. Diagnoses rendered on the limited FNA samples were compared with those made on core biopsies, resections, or cytologic samples with ample cell quantity.
Results:
DSRS of the limited FNA samples did not compromise the quality of tissues or IHC, and comparison of the limited FNA diagnosis and final diagnosis rendered on more adequate specimens revealed concordant and accurate diagnosis in 89% (8/9) of cases, in contrast to a concordance rate of 22% (2/9) before the use of DSRS while the immunostain interpretation accuracy rate is 100% (9/9).
Conclusion:
DSRS of cytology cell block sections allows IHC stains to be ascribed to particular cells, enabling diagnosis when material is sparse, when diagnostic cells are admixed with other cell types, and when the IHC stain itself would otherwise obscure the identity of cells. DSRS provides an inexpensive alternative to other, more advanced techniques such as multiplex IHC or immunofluorescence.
More Related Videos
Related Concept Videos
01:03Fixation and Sectioning
The simplest type of preparation is the wet mount, in which the specimen is placed in a drop of liquid on the slide. A liquid specimen can be directly deposited on the slide using a dropper. Solid specimens, such as skin scraping, can be placed on the slide before adding a drop of liquid to prepare the wet mount. Sometimes the liquid is simply water, but stains are often added...
01:24Simple Staining Technique
01:22Immunocytochemistry and Immunohistochemistry
These...