The DNA amount can be measured for each chromosome of the karyotype by image cytometry. Image cytometry (ICM) associates microscopy, digital image and software technologies and has been particularly useful in spatial and densitometric cytological analyses. ICM integrates the methodologies of microscopy optical calibration, standard density filters, digital CCD camera and image analysis softwares for quantitative applications. Besides of all system calibration and setup, cytological protocols must perform good slide preparations for efficient and reliable ICM analysis.