IMPROVEMENTS IN SLIDE PREPARATION FROM ARCHIVAL MATERIAL FOR AUTOMATED DNA MEASUREMENT BY IMAGE-ANALYSIS

OBJECTIVE: To optimize slide preparation for DNA content measurement b y automated image analysis and to obtain a rapid and easy method for r outine use. STUDY DESIGN: Some improvements in previously described me thods were achieved: automatic dewaxing, reduction of washing times, a ccurate adjustment of final concentration of nuclei, sedimentation and temperate drying of nuclei. RESULTS: The complete preparation of 12 s lides required four intermittent working hours. Accurate adjustment of the final concentration and sedimentation of nuclei resulted in a con stant and homogeneous distribution of nuclei. Cell loss teas prevented and the most fragile structures preserved. CONCLUSION: High-quality p reparations of nuclei lead to high-resolution histograms obtained from a large collection of events. This promotes automated image analysis method instead of flow cytometry when only archival material is availa ble for DNA content measurement and proliferating fraction evaluation.