In this study by means of acridine organge (AO) staining cell and Hadamard transform microscopic image analysis system, some factors that affect fluorescence intensity of cellular AO-DNA complex and accuracy of analytical results of human breast tumor cell nuclear DNA content were investigated. The results show that: (1) ethanol is the best fixative for cell, and aldehydes have significant negative influence on cell fluorescence; (2) cells smit stronger fluorescence when AO contains Triton X-100; (3) the optimum concentration of AO is 50μg/mL; (4) when tumor cell nulear DNA content is analyzed, satisfactory results can be obtained with either external control method (lymphocyte in blood smear or lymph node specimen smear is used as standard diploid cell) or internal control method (normal epithelial cell in the same tumor specimen is used as standard diploid cell), but the latter is preferable; (5) reliable results can be obtained when no less than sixty tumor cells are measured at random for each case.

Key words: ACRIDINE ORANGE, IMAGE ANALYSIS, MAMMARY NEOPLASMS