Time-resolved fluorescence in immunocytochemical detection of prostate-specific antigen in prostatic tissue sections
Bjartell, Anders LU ; Laine, Sari ; Pettersson, Kim ; Nilsson, Elise LU ; Lövgren, Timo and Lilja, Hans LU
(1999)
In Histochemical Journal
31(1).
p.45-52
- Abstract
Chelates with fluorescent lanthanides such as europium and terbium are widely used in immunofluorometric assays, e.g. for the measurement of different molecular forms of prostate-specific antigen (PSA) in serum for detection and monitoring of prostate cancer. These chelates have also been introduced as non-radioactive labels in immunocytochemistry and in situ hybridization. In the present study, sections of non-malignant prostate were investigated using monoclonal IgGs against PSA. Detection of specific immunostaining employing time-resolved fluorescence with europium-labeled streptavidin was compared with conventional detection by streptavidin conjugated to horse-radish peroxidase. The high PSA concentration in the tissue produced high... (More)
Chelates with fluorescent lanthanides such as europium and terbium are widely used in immunofluorometric assays, e.g. for the measurement of different molecular forms of prostate-specific antigen (PSA) in serum for detection and monitoring of prostate cancer. These chelates have also been introduced as non-radioactive labels in immunocytochemistry and in situ hybridization. In the present study, sections of non-malignant prostate were investigated using monoclonal IgGs against PSA. Detection of specific immunostaining employing time-resolved fluorescence with europium-labeled streptavidin was compared with conventional detection by streptavidin conjugated to horse-radish peroxidase. The high PSA concentration in the tissue produced high intensity, specific time-resolved fluorescence signals in the epithelial cells of the prostate gland without disturbance from non-specific tissue autofluorescence. This allowed short exposure times to be used which resulted in insignificant photobleaching. Two of the three europium-chelates evaluated yielded high signal intensities. Counterstaining was found to be optimal with Gill No. 1-Haematoxylin solution and Merckoglas(TM) was the best mounting medium for the europium chelates tested. In conclusion, time-resolved fluorescence imaging is an attractive alternative to conventional detection of streptavidin conjugated to horse-radish peroxidase, as it provides linear, high intensity, specific signals subsequent to the decay of non-specific tissue autofluorescence.
(Less)
- author
- Bjartell, Anders
LU
; Laine, Sari
; Pettersson, Kim
; Nilsson, Elise
LU
; Lövgren, Timo
and Lilja, Hans
LU
- organization
- publishing date
- 1999
- type
- Contribution to journal
- publication status
- published
- subject
- in
- Histochemical Journal
- volume
- 31
- issue
- 1
- pages
- 45 - 52
- publisher
- Springer
- external identifiers
-
- scopus:0032980411
- pmid:10405822
- ISSN
- 0018-2214
- DOI
- 10.1023/A:1003504115690
- language
- English
- LU publication?
- yes
- additional info
- Funding Information: The authors wish to thank Birgitta Frohm and Ingrid Wigheden at the Department of Clinical Chemistry, Lund University, University Hospital Malmö, Sweden, for their expert technical assistance. The study was supported by generous grants from the Biomed 2 Program, Area 4.1.7. (contract number BMH4-CT96-0453), the Swedish Medical Research Council (project number 7903), the Swedish Cancer Society (project number 3555), the Faculty of Medicine at Lund University, the Research Fund and the Cancer Research Fund at University Hospital, Malmö, the Swedish Society of Medicine (SLS), the Foundation for Urology Research in Malmö, the Magnus Bergwall Foundation, the Bergquist Foundation, Crafoord Foundation, the Gunnar, Arvid, and Elisabeth Nilsson Foundation; and the Fundacion Frederico S.A.
- id
- fe631730-8fd7-4034-8f90-71a93b016ca7
- date added to LUP
- 2022-12-06 17:42:40
- date last changed
- 2024-01-13 13:35:28
@article{fe631730-8fd7-4034-8f90-71a93b016ca7,
abstract = {{<p>Chelates with fluorescent lanthanides such as europium and terbium are widely used in immunofluorometric assays, e.g. for the measurement of different molecular forms of prostate-specific antigen (PSA) in serum for detection and monitoring of prostate cancer. These chelates have also been introduced as non-radioactive labels in immunocytochemistry and in situ hybridization. In the present study, sections of non-malignant prostate were investigated using monoclonal IgGs against PSA. Detection of specific immunostaining employing time-resolved fluorescence with europium-labeled streptavidin was compared with conventional detection by streptavidin conjugated to horse-radish peroxidase. The high PSA concentration in the tissue produced high intensity, specific time-resolved fluorescence signals in the epithelial cells of the prostate gland without disturbance from non-specific tissue autofluorescence. This allowed short exposure times to be used which resulted in insignificant photobleaching. Two of the three europium-chelates evaluated yielded high signal intensities. Counterstaining was found to be optimal with Gill No. 1-Haematoxylin solution and Merckoglas(TM) was the best mounting medium for the europium chelates tested. In conclusion, time-resolved fluorescence imaging is an attractive alternative to conventional detection of streptavidin conjugated to horse-radish peroxidase, as it provides linear, high intensity, specific signals subsequent to the decay of non-specific tissue autofluorescence.</p>}},
author = {{Bjartell, Anders and Laine, Sari and Pettersson, Kim and Nilsson, Elise and Lövgren, Timo and Lilja, Hans}},
issn = {{0018-2214}},
language = {{eng}},
number = {{1}},
pages = {{45--52}},
publisher = {{Springer}},
series = {{Histochemical Journal}},
title = {{Time-resolved fluorescence in immunocytochemical detection of prostate-specific antigen in prostatic tissue sections}},
url = {{http://dx.doi.org/10.1023/A:1003504115690}},
doi = {{10.1023/A:1003504115690}},
volume = {{31}},
year = {{1999}},
}