Improvements in slide preparation from archival material for automated DNA measurement by image analysis.
OBJECTIVE
To optimize slide preparation for DNA content measurement by automated image analysis and to obtain a rapid and easy method for routine use.


METHODS
Some improvements in previously described methods were achieved: automatic dewaxing, reduction of washing times, accurate adjustment of final concentration of nuclei, sedimentation and temperate drying of nuclei.


RESULTS
The complete preparation of 12 slides required four intermittent working hours. Accurate adjustment of the final concentration and sedimentation of nuclei resulted in a constant and homogeneous distribution of nuclei. Cell loss was prevented and the most fragile structures preserved.


CONCLUSIONS
High-quality preparations 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 available for DNA content measurement and proliferating fraction evaluation.
Duigou
F
F
Department of Pathology, Centre François Baclesse, Caen, France.
Galle
I
I
Herlin
P
P
Mandard
A M
AM
D002467:Cell Nucleus / Q000737:chemistry / Q000473:pathology; D004273:DNA, Neoplasm / Q000032:analysis*; D016591:Histocytological Preparation Techniques; D006801:Humans; D007091:Image Processing, Computer-Assisted / Q000379:methods*; D016612:Paraffin Embedding / Q000379:methods; D011003:Ploidies
D004273:DNA, Neoplasm