Related Experiment Video
Updated: Aug 11, 2026
Automated Multiplex Immunofluorescence Panel for Immuno-oncology Studies on Formalin-fixed Carcinoma Tissue Specimens
Published on: January 21, 2019
Validation of commonly used immunostains on cell-transferred cytologic specimens
Yun Gong1, Teresa Joseph, Nour Sneige
1Department of Pathology, The University of Texas M. D. Anderson Cancer Center, Houston, Texas 77030, USA. yungong@mdanderson.org
Insights
The cell-transfer technique reliably enables multiple immunocytochemical staining (ICC) on limited samples. This method shows high concordance with previous ICC results, making it a valuable tool for diagnostic pathology.
Area of Science:
- Pathology
- Immunohistochemistry
- Cytopathology
Background:
- Immunocytochemical staining (ICC) is crucial for diagnosis but often hindered by limited sample material.
- The cell-transfer technique allows dividing smear material for multiple ICC slides, addressing material limitations.
- Systematic evaluation of the reliability of ICC on cell-transferred specimens was lacking.
Purpose of the Study:
- To validate the reliability of immunocytochemical staining (ICC) performed on cell-transferred tissue specimens.
- To assess the concordance of ICC results from cell-transferred materials with previous staining results.
Main Methods:
- Validated 21 common antibodies on cell-transferred materials from 20 tumor and 2 reactive effusion samples.
- Compared ICC staining results on cell-transferred materials with prior ICC results from cell blocks or smears of the same specimens.
Main Results:
- Out of 100 immunostained transferred materials, 95 were analyzed after 5% loss.
- 97% (92/95) of cell-transferred specimens showed staining results concordant with previous ICC.
- Three discordant cases (neuroendocrine tumor, mesothelioma, metastatic lung carcinoma) showed negative results on cell-transferred material where prior ICC was focally positive.
Conclusions:
- Immunocytochemical staining (ICC) is generally reliable on cell-transferred tissue specimens for the tested markers.
- Caution is advised when interpreting ICC results from cell-transferred specimens with limited cellularity or heavy background staining.
Background:
Immunocytochemical staining (ICC) is often limited by the lack of cell blocks that contain diagnostic cells or by the unavailability of smear materials. The cell-transfer technique, in which original smear material is divided into several pieces and then transferred to multiple slides, can facilitate multiple ICC on limited materials. However the reliability of the staining results has not been systematically evaluated.
Methods:
The authors validated 21 commonly used antibodies (cytokeratin [CK] 7, CK 20, TTF-1, panCK, vimentin, HMB-45, Mart-1, leukocyte common antigen, chromogranin, synaptophysin, estrogen receptor, progesterone receptor, prostate-specific antigen, prostatic acid phosphatase, calretinin, CK5/6, WT-1, BerEP4, MOC-31, p63, and thyroglobulin) on the cell-transferred materials of various tumor samples (n = 20) and reactive effusion samples (n = 2). The staining results were compared with the previously available ICC results (performed either on cell block sections or on smears) of the same specimens.
Results:
One hundred pieces of transferred materials were immunostained, of which 5 (5%) were lost during the staining procedure. Of the remaining 95 pieces, 92 (97%) showed staining results that agreed with those of the previous ICC, whereas 3 (3%) showed results that disagreed. The original ICC in the latter three specimens was performed on cell block sections and showed focally positive synaptophysin staining in a sample with a neuroendocrine tumor, focally positive CK5/6 staining in a pleural effusion sample with mesothelioma, and positive TTF-1 staining in a lymph node sample with metastatic lung carcinoma. Their corresponding ICC of cell-transferred materials showed negative results. An increase in background staining was observed in 1 of the 95 tissue pieces, but no false-positive results were observed.
Conclusions:
In general, ICC can be reliably performed on cell-transferred tissue specimens for markers tested. However, caution should be taken when interpreting ICC results in tissue specimens with limited cellularity and heavy background staining.
