Ed. Spots were accepted (counted) if they had a minimum distance
Ed. Spots were accepted (counted) if they had a minimum distance of 0.5 m and a minimum intensity of 27 compared to the brightest spot in the same PubMed ID:https://www.ncbi.nlm.nih.gov/pubmed/25636517 cell. Reject if > 2 Cells with more than 2 red spots were automatically rejected. SpotCounts (14,78) The number of green FISH spots was determined. Spots were accepted (counted) if they had a minimum distance of 1.4 m and a minimum intensity of 78 compared to the brightest spot in the same cell. Reject if > 2 Cells with more than 2 green spots are automatically rejected. Yes Cells not showing any X signals are automatically rejected.100/15/Most important parameters for the classifier used for analyzing the patient samples.1,1-Dimethylbiguanide hydrochloride molecular weight ConclusionAutomated chimerism analysis is a robust and sensitive method which can be used in routine diagnosis to detect residual cells effectively and economically. Simultaneous hybridization of gonosomes and disease-marker represent a sensitive method to detect disease-marker positive residual cells with a very low cut off level. The amount of residual cells correlates with survival. There are patients with residual cells < 4 without tendency of relapse. The detection of disease-marker positive residual cells up to 1.3 does not correlate with relapse. Disease-marker positive and disease-marker negative residual cells can appear at the same time in one sample. The definite origin of disease-marker negative residual cells is unclear and should be investigated in a large multicenter study.row transplantation (BMT). The median age of the transplanted patients was 46 years (2?9 years). 12 patients showed cytogenetic disease-marker in their malignant cells, i.e. a bcr/abl-fusion in nine and a trisomy 8 in three patients. Overall, 19 patients died, either due to relapse (n = 9), a graft-versus-host-disease (n = 3) or an infection (n = 4). In 3 patients the reason of death remained unclear.Cytogenetics and molecular cytogenetics including FISH analysis Standard techniques were used to cultivate leukocytes out of venous blood, prepare chromosome-preparations [43], and to perform interphase FISH analysis [44]. Commercially available probes (Abbott, Wiesbaden, Germany) for LSI-ES bcr/abl, centromere 8, X and Y were applied. Automatic chimerism analysis For automated analysis we used an Axioplan 2 Imaging microscope (Carl Zeiss Jena, Germany) equiped with CCD-camera CV-M1, 1280 ?1024 pixel (Jai Glostrup, Denmark) and a motorized stage with 8 slide positions (M zh ser, Wetzlar, Germany). All components were connected to a personal computer (Dell, Langen, Germany) running the Metafer/MetaCyte-Software from MetaSystems (Altlussheim, Germany).MethodsControls Peripheral blood samples of 21 clinically healthy male (11) and female (10) between 6 and 67 years were studied as controls. Patients A total of 28 female and 30 male patients were analyzed retrospectively after sex-mismatched stem cell transplantation which were performed between 1995 and 2006 at the University Medical Centre Jena. As shown in Table 1, there were 24 acute myloid leukemia (AML), 16 chronic myeloid leukemia (CML), 5 acute lymphatic leukemia (ALL), 4 multiple myeloma (MM), 3 severe aplastic anemia (SAA), 2 myelodysplastic syndrome (MDS), and 1 patient each with Non-Hodgkin lymphoma (NHL), chronic lymphatic leukemia (CLL), lymphoma (Lym) and osteopetrosis (OP). Conditioning regimens were dose reduced in 30 patients or myeloablative in 23 patients [44-46] 54 patients underwent peripheral blood stem-cell transplantatio.