Introduction: In effusion cytology, immunocytochemistry is a useful staining approach to provide important information for diagnosis. Effusion cytology is performed not only for pleural effusions and ascites but also for peritoneal and needle washing from fine needle aspirations or instruments. Although various solutions are used for washing cytology, the effect of the solution type on immunocytochemical reactivity is not fully understood. In this study, we examined the immunocytochemical reactivity of cytological samples after storage in various solutions. Methods: Cell block specimens were obtained from ascites of patients with peritoneal cancer and pleural effusions of patients with diffuse malignant mesothelioma. Various solutions, including physiological saline (PS), Ringer solution, a low-molecular-weight dextran L injection, Voluven 6% solution, Mixid L injection, RPMI-1640 medium, and horse serum were added to the sediment layers of aliquots. All samples were kept at 4°C, and aliquots were subsequently processed at specific time points (0, 1, 2, 4, 7, and 14 days). Formalin-fixed, paraffin-embedded, cell block samples were prepared for immunocytochemical staining. Immunocytochemical results were analyzed for differences in the percentages of positive cells, using the effusion sample stored for 1 h as standard (100%). Results: For all solutions other than PS, the median and central 50% of values were <100% (with respect to the effusion sample as a standard) after 1 h of storage. Immunoreactivity decreased for most solutions as time progressed. Conclusion: Of note, immunocytochemistry results obtained using a washing solution are different from those using an effusion sample. For cytology, when a washing solution was used or when a sample was stored for a long time, the accuracy of the immunocytochemical results was low.

1.
Engels
M
,
Michael
C
,
Dobra
K
,
Hjerpe
A
,
Fassina
A
,
Firat
P
.
Management of cytological material, pre-analytical procedures and bio-banking in effusion cytopathology
.
Cytopathology
.
2019
Jan
;
30
(
1
):
31
8
.
[PubMed]
0956-5507
2.
Ikeda
K
,
Tate
G
,
Suzuki
T
,
Mitsuya
T
.
Comparison of immunocytochemical sensitivity between formalin-fixed and alcohol-fixed specimens reveals the diagnostic value of alcohol-fixed cytocentrifuged preparations in malignant effusion cytology
.
Am J Clin Pathol
.
2011
Dec
;
136
(
6
):
934
42
.
[PubMed]
0002-9173
3.
Ikeda
K
,
Tate
G
,
Suzuki
T
,
Kitamura
T
,
Mitsuya
T
.
Diagnostic usefulness of EMA, IMP3, and GLUT-1 for the immunocytochemical distinction of malignant cells from reactive mesothelial cells in effusion cytology using cytospin preparations
.
Diagn Cytopathol
.
2011
Jun
;
39
(
6
):
395
401
.
[PubMed]
8755-1039
4.
Ikeda
K
,
Tate
G
,
Suzuki
T
,
Kitamura
T
,
Mitsuya
T
.
IMP3/L523S, a novel immunocytochemical marker that distinguishes benign and malignant cells: the expression profiles of IMP3/L523S in effusion cytology
.
Hum Pathol
.
2010
May
;
41
(
5
):
745
50
.
[PubMed]
0046-8177
5.
Renshaw
SA
,
Gupta
S
,
Campos
M
,
Hodes
L
,
Renshaw
AA
,
Gould
EW
.
The addition of RPMI significantly improves the cellularity of cerebrospinal fluid cytology specimens over time.
Cancer Cytopathol. May;121(5):271–4.
6.
Manosca
F
,
Schinstine
M
,
Fetsch
PA
,
Sorbara
L
,
Maria Wilder
A
,
Brosky
K
, et al.
Diagnostic effects of prolonged storage on fresh effusion samples
.
Diagn Cytopathol
.
2007
Jan
;
35
(
1
):
6
11
.
[PubMed]
8755-1039
7.
Michael
CW
,
Davidson
B
.
Pre-analytical issues in effusion cytology
.
Pleura Peritoneum
.
2016
Mar
;
1
(
1
):
45
56
.
[PubMed]
2364-768X
8.
Antonangelo
L
,
Vargas
FS
,
Acencio
MM
,
Corá
AP
,
Teixeira
LR
,
Genofre
EH
, et al.
Effect of temperature and storage time on cellular analysis of fresh pleural fluid samples
.
Cytopathology
.
2012
Apr
;
23
(
2
):
103
7
.
[PubMed]
0956-5507
9.
Ikeda
K
,
Sato
S
,
Chigira
H
,
Shibuki
Y
,
Hiraoka
N
.
Characterizing the effect of automated cell sorting solutions on cytomorphological changes
.
Acta Cytol
.
2019
Jun
;
24
:
1
9
.
[PubMed]
0001-5547
10.
Risberg
B
,
Davidson
B
,
Dong
HP
,
Nesland
JM
,
Berner
A
.
Flow cytometric immunophenotyping of serous effusions and peritoneal washings: comparison with immunocytochemistry and morphological findings
.
J Clin Pathol
.
2000
Jul
;
53
(
7
):
513
7
.
[PubMed]
0021-9746
11.
Guzman
J
,
Arbogast
S
,
Bross
KJ
,
Finke
R
,
Costabel
U
.
Effect of storage time of pleural effusions on immunocytochemical cell surface analysis of tumor cells
.
Anal Quant Cytol Histol
.
1992
Jun
;
14
(
3
):
203
9
.
[PubMed]
0884-6812
12.
Guzman
J
,
Arbogast
S
,
Bross
KJ
,
Finke
R
,
Costabel
U
.
Effect of storage time on the analysis of lymphocyte subpopulations in pleural effusions
.
Acta Cytol
.
1993
May-Jun
;
37
(
3
):
267
71
.
[PubMed]
0001-5547
Copyright / Drug Dosage / Disclaimer
Copyright: All rights reserved. No part of this publication may be translated into other languages, reproduced or utilized in any form or by any means, electronic or mechanical, including photocopying, recording, microcopying, or by any information storage and retrieval system, without permission in writing from the publisher.
Drug Dosage: The authors and the publisher have exerted every effort to ensure that drug selection and dosage set forth in this text are in accord with current recommendations and practice at the time of publication. However, in view of ongoing research, changes in government regulations, and the constant flow of information relating to drug therapy and drug reactions, the reader is urged to check the package insert for each drug for any changes in indications and dosage and for added warnings and precautions. This is particularly important when the recommended agent is a new and/or infrequently employed drug.
Disclaimer: The statements, opinions and data contained in this publication are solely those of the individual authors and contributors and not of the publishers and the editor(s). The appearance of advertisements or/and product references in the publication is not a warranty, endorsement, or approval of the products or services advertised or of their effectiveness, quality or safety. The publisher and the editor(s) disclaim responsibility for any injury to persons or property resulting from any ideas, methods, instructions or products referred to in the content or advertisements.
You do not currently have access to this content.