Karyotype of ten fibroblasts after Alu sequences and telomeres hybridization

The image bellow  was obtain after sequential hybridization of telomeric PNA probe (FITC:green) followed by hybridization of Alu-PCR product (Rhodamin: Red):

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:

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:

Karyotyping:

Combining R bands (Alu), G bands (DAPI) and chromosomal painting allows to classify chromosomes in a karyogram:

Left:R Bands (Alu sequences). Right: G bands (Inverse DAPI+DoG filter).