Automated microscopy of amniotic fluid cells: detection of FISH signals using the FastFISH imaging system.
OBJECTIVE
FISH (fluorescence in situ hybridization) analysis is a valuable adjunct to cytogenetics that provides a rapid screen for common abnormalities. However, FISH is expensive, labor-intensive, and requires a high skill level and subjective signal interpretation. A fully automated system for FISH analysis could improve laboratory efficiency and potentially reduce errors and costs.


METHODS
In this study we blindly compared automated FISH signal acquisition and display against standard FISH analysis. A total of 62 amniocentesis samples were prepared using the AneuVysion multicolor DNA probe kit and probed for chromosomes 13, 18, 21, X, and Y. Two sets of slides were produced from each sample. Fifty cells were scored in each slide. One set was evaluated using standard manual microscopy and the other using the automated image acquisition and display capabilities of the Ikoniscope fastFISH amnio Test System. This system uses epifluorescence optics, along with optimized slide management to process slides automatically.


RESULTS
A 100% concordance was observed between the results obtained using manual microscopy and the automated system. There was also 100% concordance between the FISH results and those obtained by conventional karyotyping.


CONCLUSIONS
Our data suggest that the automated system is capable of providing accurate and rapid identification and display of cells and FISH signals.
Evans
Mark I
MI
Comprehensive Genetics, New York, NY 10021, USA. Evans@CompreGen.com
Sharp
Michael
M
Tepperberg
James
J
Kilpatrick
Michael W
MW
Tsipouras
Petros
P
Tafas
Triantafyllos
T
D000293:Adolescent; D000328:Adult; D000649:Amniocentesis; D025063:Chromosome Disorders / Q000175:diagnosis*; D005260:Female; D006801:Humans; D007091:Image Processing, Computer-Assisted / Q000295:instrumentation; D017404:In Situ Hybridization, Fluorescence / Q000379:methods*; D007621:Karyotyping / Q000379:methods; D008856:Microscopy, Fluorescence / Q000295:instrumentation; D008875:Middle Aged; D011247:Pregnancy; D012680:Sensitivity and Specificity