Analysis of the reliability of manual and automated immunohistochemical staining procedures. A pilot study.
OBJECTIVE
To study the variation in the number of stained cells and staining intensity comparing 2 immunostainers and manual staining for estrogen receptor (ER) expression in breast carcinoma.


METHODS
In 5 cases, 15 consecutive paraffin sections were investigated after simultaneous immunohistochemical ER staining. The slides were evaluated using a CM-2 TV image analysis system (Hund, Wetzlar, Germany). One viewing field, identified around a histologic structure present on all 15 sections, was analyzed. The percentage of immunoreactive cells (PP), mean grey values of the immunopositive (GVpos.) and immunonegative nuclei (GVneg.), and immunohistochemical staining intensity (SI, defined as GVneg.-GVpos.) were calculated.


RESULTS
The mean PP values were higher for immunostainers A (70.2%) and B (53.8%) than for manual staining (40.8%). The results were significantly different comparing the 2 immunostainers (P = .0143) or immunostainer A and manual staining (P < .0001). Also, the mean SI values were higher for immunostainers A (24.5 +/- 2.8% [CV]) and B (18.5 +/- 31.1%) than for manual staining (10.8 +/- 33.8%). These differences revealed statistical significance comparing the immunostainers with manual staining (.0001 < P = .0048).


CONCLUSIONS
Our results underline the higher staining quality using immunostainers in comparison with manual staining.
Department of Pathology, Technical University of Aachen, Pauwelsstrasse 30, 52057 Aachen, Federal Republic of Germany. biesterfeld@pathologie-re.de
Stefan
S
Biesterfeld
Helen Louise
HL
Kraus
Thorsten
T
Reineke
Leon
L
Muys
Ana Maria
AM
Mihalcea
Christian
C
Rudlowski
D001331:Automation; D001943:Breast Neoplasms / Q000473:pathology; D005260:Female; D006801:Humans; D007150:Immunohistochemistry / Q000295:instrumentation* / Q000379:methods / Q000592:standards; D011960:Receptors, Estrogen / Q000032:analysis; D015203:Reproducibility of Results; D013194:Staining and Labeling / Q000295:instrumentation* / Q000379:methods / Q000592:standards
D011960:Receptors, Estrogen