Automated analysis of multiple sections for the detection of occult cells in lymph nodes.
OBJECTIVE
At present, reverse transcription (RT)-PCR against carcino-embryonic antigen mRNA is one of the few research tools for the detection of occult cells in histopathologically assessed negative lymph nodes from patients with colorectal cancer. The aim of this study was to investigate the suitability of supervised low-resolution image analysis of immunohistochemically stained sections as alternative.


METHODS
Multiple sections (n = 50) of regional lymph nodes from patients with colorectal cancer were immunohistochemically stained and analyzed by applying low-resolution image analysis (flatbed scanning) for semiautomated detection of cytokeratin (CK)-positive stained cells. The sensitivity of this approach was demonstrated for 20 patients with stage II colorectal cancer and compared with RT-PCR regarding the detection of clinically assessed recurrence of disease within 10 years.


RESULTS
CK(+) cells were detected in all of the patients (n = 6; 100%) with recurrence, compared with five patients (83%) found positive by carcinoembryonic antigen RT-PCR. From patients (n = 14) who did not develop a recurrence, eight (57%) had positive lymph nodes. In all patients with recurrence, we visually identified at least one group of CK(+) cells (>/==" BORDER="0">2 cells).


CONCLUSIONS
Automated image analysis is a promising tool for the detection of occult cells in histopathologically negative nodes. It is potentially more sensitive but less specific for detecting recurrence of disease than conventional histopathology or RT-PCR and is particularly useful for the evaluation of sentinel nodes. Furthermore, it opens new ways for basic research of occult cells based on molecular profiling after laser-microdissection.
Laboratory for Cytochemistry and Cytometry, Department of Molecular Cell Biology, Leiden University Medical Center, The Netherlands. W.E.Mesker@LUMC.NL
Wilma E
WE
Mesker
Fania S
FS
Doekhie
Hans
H
Vrolijk
Rob
R
Keyzer
Willem C R
WC
Sloos
Hans
H
Morreau
Paddy S
PS
O'Kelly
Geertruida H
GH
de Bock
Rob A E M
RA
Tollenaar
Hans J
HJ
Tanke
D001331:Automation; D001671:Biochemistry / Q000379:methods*; D002272:Carcinoembryonic Antigen / Q000096:biosynthesis; D015179:Colorectal Neoplasms / Q000175:diagnosis / Q000473:pathology; D006801:Humans; D007091:Image Processing, Computer-Assisted; D007150:Immunohistochemistry; D007633:Keratins / Q000096:biosynthesis; D008198:Lymph Nodes / Q000473:pathology*; D008207:Lymphatic Metastasis; D009382:Neoplasms, Unknown Primary / Q000473:pathology*; D012333:RNA, Messenger / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012680:Sensitivity and Specificity
D002272:Carcinoembryonic Antigen; D012333:RNA, Messenger; D007633:Keratins