TM-FISH Fig 01 - legend

Fig. 1a-c. TM-FISH analysis of a case with 46,XX add(1)(1q41), based on the 1/5 detection scheme. Using Photoshop (Adobe), grayscale images of the same metaphase in FITC (F), rhodamine-Cy3 (R), DNP-Cy3.5 (N), BIO-Cy5 (B), DIG-DEAC (D) and DAPI channels were combined and pseudocolored into two RGB images : FR (including FITC-green, Rhodamine-red and DAPI-blue) and NBD (including DNP/Cy3.5-red, BIO/Cy5-blue and DIG/DEAC-green). The arrows in a and b indicate the extra material on 1qter with a fluorescent signal carrying the color signature of chromosome 8 telomeres. Numbers in a show the position of all human chromosomes. The two small arrows in the upper right corner point to the same translocation as detected by M-FISH (very weak signal) or G-banding (the q arm of human chromosome 1). See main text for details. For a better signal visualization, in c, the image of one chromosome of every human pair was split in its original grayscale images. The vertical lines separate the FR from NBD images. The first chromosome in every group as well as the first chromosome after the vertical white line depicts the merged color image, as it appears in figures a and b. The white circle indicates the DAPI image of that chromosome. The red, green or blue dots above each chromosome indicate the pseudo-color assigned for that channel in the respective RGB image (FR or NBD). The same legend appears in the bottom right corner of c. The color chart in d depicts the expected color of every human subtelomeric probe in the FR and NBD images respectively, and also indicates (letters) what haptenes (see discussion above) were used to label each telomere. For example, chromosome 1 telomeric probes were colored red in the FR image and magenta in NBD, indicating that they were labeled by rhodamine (R), DNP (N) and BIO (B). The 2/3 detection scheme (not shown) can be displayed the same way as in a and b, the only difference being that the two images, FR and NBD, come from different metaphases. The same chart (d) can be used for the mouse TM-FISH, with some exceptions, also shown in Table 1. Thus, mouse chromosome 5 was labeled FNB (green in the FR and magenta in the NBD images - similar to human 21); chromosome 6 was labeled R (red in the FR image - similar to human 5) and X was labeled N (red in the NBD image - similar to human 20). No probes for the mouse Y chromosome were used.