Skip to main content

Lund University Publications

LUND UNIVERSITY LIBRARIES

The impact of different fixatives on immunostaining of lung adenocarcinomas in pleural effusion cell blocks

Mansour, Mohammed S I LU orcid ; Pettersson, Louise LU orcid ; Seidal, Tomas ; Strömberg, Ulf ; Mager, Ulrich ; Ali, Lana ; Kumbaric, Sana ; Hejny, Kim ; Taheri-Eilagh, Fereshteh and Mufti, Joudy , et al. (2024) In Cancer Cytopathology
Abstract

BACKGROUND: Cell blocks (CBs) are widely used for biomarker analyses such as immunostaining. Although immunohistochemistry on formalin-fixed paraffin-embedded tissues is standardized, there are multiple preparation methods and fixatives for cytology. Our objective was to investigate the effect of different common fixatives on the immunoreactivity of pleural effusion CBs with metastatic lung adenocarcinomas.

METHODS: This prospective study included 24 malignant pleural effusions from different patients with lung adenocarcinoma. From each case, four identical CBs were fixed in 10% neutral buffered formalin, PreservCyt, CytoLyt, and CytoRich Red (only 17 of the cases), respectively. Samples containing <100 malignant cells were... (More)

BACKGROUND: Cell blocks (CBs) are widely used for biomarker analyses such as immunostaining. Although immunohistochemistry on formalin-fixed paraffin-embedded tissues is standardized, there are multiple preparation methods and fixatives for cytology. Our objective was to investigate the effect of different common fixatives on the immunoreactivity of pleural effusion CBs with metastatic lung adenocarcinomas.

METHODS: This prospective study included 24 malignant pleural effusions from different patients with lung adenocarcinoma. From each case, four identical CBs were fixed in 10% neutral buffered formalin, PreservCyt, CytoLyt, and CytoRich Red (only 17 of the cases), respectively. Samples containing <100 malignant cells were excluded. All CBs were stained with thyroid transcription factor 1 (TTF-1; clones 8G7G3/1 and SPT24), napsin A, claudin 4, CEA, CK7, and epithelial cell adhesion molecule (EpCAM; clones BS14, Ber-Ep4, and MOC-31). The fraction and intensity of stained cells were evaluated.

RESULTS: Of the investigated markers, a significant difference in staining proportion was seen for TTF-1 clone 8G7G3/1 and EpCAM clone MOC-31, especially with cases being negative in CytoLyt (33.3% and 83.3% positive, respectively) and PreservCyt (62.5% and 83.3%) whereas being positive in CytoRich Red (76.5% and 94.1%) and formalin (both 95.8%). A significantly weaker intensity of staining was seen for all alcohol-based fixatives compared to formalin for TTF-1 clone 8G7G3/1, napsin A, and EpCAM clone MOC-31, whereas EpCAM clone Ber-Ep4 was significantly weaker only in PreservCyt compared with formalin.

CONCLUSIONS: Immunocytochemical expression and concordance with formalin-fixed CBs differ depending on the used fixative as well as the antibody and clone, warranting investigation of the reliability of each biomarker for non-formalin-fixed cytology.

(Less)
Please use this url to cite or link to this publication:
author
; ; ; ; ; ; ; ; and , et al. (More)
; ; ; ; ; ; ; ; ; ; and (Less)
organization
publishing date
type
Contribution to journal
publication status
epub
subject
in
Cancer Cytopathology
publisher
Wiley-Blackwell
external identifiers
  • pmid:38829774
  • scopus:85194901823
ISSN
1934-6638
DOI
10.1002/cncy.22833
project
Biomarkers in mesothelioma and non-small cell lung cancer: Investigation of cytological specimens with correlation to histology
Improvement of diagnostic and prognostic cytopathology of lung cancer and malignant mesothelioma.
language
English
LU publication?
yes
additional info
© 2024 The Authors. Cancer Cytopathology published by Wiley Periodicals LLC on behalf of American Cancer Society.
id
b0ee80fa-32bb-4845-8248-6df65640ac34
date added to LUP
2024-08-06 18:49:21
date last changed
2024-08-21 05:21:29
@article{b0ee80fa-32bb-4845-8248-6df65640ac34,
  abstract     = {{<p>BACKGROUND: Cell blocks (CBs) are widely used for biomarker analyses such as immunostaining. Although immunohistochemistry on formalin-fixed paraffin-embedded tissues is standardized, there are multiple preparation methods and fixatives for cytology. Our objective was to investigate the effect of different common fixatives on the immunoreactivity of pleural effusion CBs with metastatic lung adenocarcinomas.</p><p>METHODS: This prospective study included 24 malignant pleural effusions from different patients with lung adenocarcinoma. From each case, four identical CBs were fixed in 10% neutral buffered formalin, PreservCyt, CytoLyt, and CytoRich Red (only 17 of the cases), respectively. Samples containing &lt;100 malignant cells were excluded. All CBs were stained with thyroid transcription factor 1 (TTF-1; clones 8G7G3/1 and SPT24), napsin A, claudin 4, CEA, CK7, and epithelial cell adhesion molecule (EpCAM; clones BS14, Ber-Ep4, and MOC-31). The fraction and intensity of stained cells were evaluated.</p><p>RESULTS: Of the investigated markers, a significant difference in staining proportion was seen for TTF-1 clone 8G7G3/1 and EpCAM clone MOC-31, especially with cases being negative in CytoLyt (33.3% and 83.3% positive, respectively) and PreservCyt (62.5% and 83.3%) whereas being positive in CytoRich Red (76.5% and 94.1%) and formalin (both 95.8%). A significantly weaker intensity of staining was seen for all alcohol-based fixatives compared to formalin for TTF-1 clone 8G7G3/1, napsin A, and EpCAM clone MOC-31, whereas EpCAM clone Ber-Ep4 was significantly weaker only in PreservCyt compared with formalin.</p><p>CONCLUSIONS: Immunocytochemical expression and concordance with formalin-fixed CBs differ depending on the used fixative as well as the antibody and clone, warranting investigation of the reliability of each biomarker for non-formalin-fixed cytology.</p>}},
  author       = {{Mansour, Mohammed S I and Pettersson, Louise and Seidal, Tomas and Strömberg, Ulf and Mager, Ulrich and Ali, Lana and Kumbaric, Sana and Hejny, Kim and Taheri-Eilagh, Fereshteh and Mufti, Joudy and Nakdali, Dawla and Brunnström, Hans}},
  issn         = {{1934-6638}},
  language     = {{eng}},
  month        = {{06}},
  publisher    = {{Wiley-Blackwell}},
  series       = {{Cancer Cytopathology}},
  title        = {{The impact of different fixatives on immunostaining of lung adenocarcinomas in pleural effusion cell blocks}},
  url          = {{http://dx.doi.org/10.1002/cncy.22833}},
  doi          = {{10.1002/cncy.22833}},
  year         = {{2024}},
}