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The number of cells was estimated by optical fractionator probe using a computer assisted image analysis system with a Zeiss Axioplan 2 microscope connected to a digital camera.
The images were analyzed using a computer assisted image analyzer system consisting of a microscope (Olympus CX-41 Tokyo, Japan) equipped with a high-resolution video camera (Olympus DP21, Japan).
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Colonies with more than 50 cells were counted using digital images of Petri dishes containing fixed and stained clones, associated with a computer assisted image analysis system (Samba Technologies, Meylan, France) as previously described (Biston et al, 2003), which allows to avoid subjective bias in the procedure.
Then, after staining with alkaline phosphatase, the spots in each well were counted using a computer assisted video image analysis system (Zeiss Co., Germany).
Microscopic images were transferred to a monitor (PVM 1444QM; Sony Corp., Tokyo, Japan) by a low light camera (Kappa CF 8/1; Kappa Messtechnik, Gleichen, Germany) and recorded on a video tape for later evaluation using a computer assisted system for microcirculation analysis (Cap image; Zeintl, Heidelberg, Germany).
Images were analysed using a computer assisted method (Byng et al, 1994, 1996a, b).
The qualitative data analysis was undertaken using a computer assisted qualitative data analysis software: NVivo©.
Trained interviewers interviewed each case and control using a computer assisted telephone interview.
Trained interviewers carried out interviews using a computer assisted personal interviewing (CAPI) system.
Treated semen parameters were analyzed using a computer assisted semen analyzer (CASA).
Statistical analysis was performed on results using a computer assisted statistical program (Epi Info, revision 6.04 b, CDC, USA).
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