1. SAMPLE PREPARATION
Tissue samples were frozen in liquid nitrogen for transport and conservation until the time of analysis. They were mechanically disaggregated in citrate buffer pH 7.6. The tissue was finally chopped and squeezed with tweezers. 1-2 10^6 cells/nuclei were centrifuged at 300xg for 10 minutes. Once the supernatant was aspirated, the pellet was subjected to DNA staining acording to Vindelöv and Christensen technique (Cytometry 11, 753-770, 1990) .
2. ANALYSIS CONDITIONS
A FACScan cytometer from Becton Dickinson was used. The signal was collected in the FL2 detector at 585 nm. 10-15,000 events per sample were collected. The instrument linearity was check using chicken erithrocyte nuclei (DNA QC Particles, Becton Dickinson) and the diploid channel was adjusted using a diploid colon sample.
The Cell Cycle and Ploidy analysis were evaluated using "Multicycle Autofit Ver.2.50 from Phoenix Flow Systems, San Diego, CA" software.
3. CLINICAL DATA
Each case is accompanied by a brief summary of its clinical history.
Case 1
Case 2
Case 3
Case 4
Case 5
Case 6
Case 7
Case 8
Case 9
Case 10
Tlf. 510 36 59 - 510 36 60
e-mail: andres@imago.quimica.uniovi.es