Myeloid Leukemia

Chimerism Analysis 285
Fig. 2. Calculation of chimerism in percent recipient T-cells. A–D show paradigms
for the calculation of chimerism. In the formulas indicated, the letters A–D represent
either peak heights or areas (see Note 16). Alleles shared between donor and recipient
(C, allele B) are disregarded in the calculation. Abbreviations: Don = Donor; Rec =
Recipient; Chim = Chimeric situation.
Microsatellites for initial genotyping and selection of markers for the follow-up
of chimerism are described in Table 1 and Notes 21–24.
3.4. FISH Analysis of X and Y Chromosomes for Chimerism Analysis
in the Sex-Mismatched Transplant Setting (see Fig. 3)
3.4.1. Application of Cells to the Slide(s)
1. Centrifuge cell suspension (e.g., flow-sorted cell populations) in Eppendorf tubes
for 10 min at approx 4000g.
2. Remove supernatant, leaving about 5 µL liquid in the tube.
3. Resuspend cells and apply the suspension to an appropriately labeled chamber on
the slide.
4. Air-dry the slide(s) at room temperature (RT) for at least 2 h.