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6th ESACP Congress, Heidelberg, April 7-11, 1999 |
A101
The Laser Scanning Cytometer (LSC) enables multicolor fluorescence and light
scatter measurements on a slide featuring relocation of single cells for
further investigation and the ability to confirm and capture investigated
cells via a CCD camera.
März H, Baumgartner A, Hambsch J, Nüsse M, Schmid T,
Schneider P, Zotz R, Tarnok A
Heart Center Leipzig GMBH - University of Leipzig,
Pediatric Cardiology, Leipzig, Germany
The use of flow cytometry (FCM) is limited in certain cases of FISH assays.
While investigating sperm of human and mice aneuploid chromosomes are
identified due to the amounts of FISH spots. Since FCM only detects integrated
fluorescence signals of single cells, it cannot provide the resolution which
is therefore demanded while the LSC can do. Due to its technique, the Laser
Scanning Cytometer enables to detect immobilised cell specimen on a slide.
Therefore, it provides a distinguishable distribution of FISH spots on sperm which
is applied for an automated scoring technique.
In relation to time-consuming manual scoring using a fluorescence microscope,
the LSC enabled to determine more than 15,000 sperm in about 30 minutes.
Furthermore, we took advantage of another LSC feature, automatic cell cycle
analysis. This feature is directly supported by the software but due to the
relocation after the measurement it is also possible to validate the
correlated cells preventing false positive events in calculation of the
apoptosis index. In the next future, we are aiming to establish the LSC for
our clinical diagnostic facilities for neonates undergoing heart surgery.
Therefore, we hope to reduce the demanded blood sample volume from 400
µl to 5-10 µl. For the moment, we succeeded to minimize the needed volume
to approximately 100 µl.