The image bellow was obtain after sequential hybridization of telomeric PNA probe (FITC:green) followed by hybridization of Alu-PCR product (Rhodamin: Red):
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| Pairs of chromosomes are ordered by columns from HSA 1 (left) to XY (right) . Metaphases are ordered by row. |
Image acquisition was performed with a low resolution 8bits camera mounted on a Leica DMRB fluorescence microscope (100x). Raw images were transferred from a Unix Cytovision station with 3 inch 1/2 floppy disk to a power Macintosh 9500.
Example of a metaphase after alignment of Alu image on DAPI / telomeres images:
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| Alu images were aligned on DAPI by hand using Photoshop 3 (Here no background correction, nor contrast enhancement)
Whole chromosome painting was also performed sequentially, simultaneously on HSA-13 and HSA-6 and HSA-X:
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Karyotyping:
Combining R bands (Alu), G bands (DAPI) and chromosomal painting allows to classify chromosomes in a karyogram:
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| Left:R Bands (Alu sequences). Right: G bands (Inverse DAPI+DoG filter). |
