9 Human Sperm Cells
135
Comments
FISH analysis on interphase nuclei generally suffers from systematic errors. This means, that the number of detected signals varies from the number of actual DNA binding sites. The signal number may be artificially
decreased by:
- insufficient tissue penetrability resulting in minor hybridization efficiency,
- a certain amount of two actual signals lying so close together that they
appear as one,
- analysis of damaged or overlapping cells,
or increased by:
- signal splitting due to dispersed DNA
- signal imitating by bacteria or other background fluorescence.
False negative signals from insufficient tissue penetrability may be reduced by optimal tissue pretreatment, while the fact that a certain number
of signals stick together, simulating a single signal has to be addressed by a
statistical approach. False positive signals by background fluorescence can
be reduced by working as cleanly as possible, by optimized washing procedures or by the use of direct labeled DNA probes.
Applications
FISH analyses of human sperm cells have been applied for basic studies of
aneuploidy (Martin et al. 1995, Hu et al. 1996, Martin et al. 1996, Spriggs et
al. 1996, Rademaker et al. 1997, Mennicke et al. 1997), to study chromosomal aberrations in infertile men (Moosani et al. 1995, Martin 1996, Lahdetie et al. 1997) and men exposed to potential mutagens (Martin and
Barclay 1996, Martin 1997, Martin et al. 1997, Robbins et al. 1997, Robbins
et al. 1997, Harkonen et al. 1999, Sram et al. 1999) as well as to analyze the
frequency of structural abnormal chromosomes (Vanhummelen et al.
1996, Pfeiffer et al. 1998, Martini et al. 1998, Colls et al. 1998, Baumgartner
et al. 1999, Durak et al. 1999).
Acknowledgements. I am grateful to Dr. U. Trautmann and Dr. P. Binder-Wirth for sharing
their technical experience.
135
Comments
FISH analysis on interphase nuclei generally suffers from systematic errors. This means, that the number of detected signals varies from the number of actual DNA binding sites. The signal number may be artificially
decreased by:
- insufficient tissue penetrability resulting in minor hybridization efficiency,
- a certain amount of two actual signals lying so close together that they
appear as one,
- analysis of damaged or overlapping cells,
or increased by:
- signal splitting due to dispersed DNA
- signal imitating by bacteria or other background fluorescence.
False negative signals from insufficient tissue penetrability may be reduced by optimal tissue pretreatment, while the fact that a certain number
of signals stick together, simulating a single signal has to be addressed by a
statistical approach. False positive signals by background fluorescence can
be reduced by working as cleanly as possible, by optimized washing procedures or by the use of direct labeled DNA probes.
Applications
FISH analyses of human sperm cells have been applied for basic studies of
aneuploidy (Martin et al. 1995, Hu et al. 1996, Martin et al. 1996, Spriggs et
al. 1996, Rademaker et al. 1997, Mennicke et al. 1997), to study chromosomal aberrations in infertile men (Moosani et al. 1995, Martin 1996, Lahdetie et al. 1997) and men exposed to potential mutagens (Martin and
Barclay 1996, Martin 1997, Martin et al. 1997, Robbins et al. 1997, Robbins
et al. 1997, Harkonen et al. 1999, Sram et al. 1999) as well as to analyze the
frequency of structural abnormal chromosomes (Vanhummelen et al.
1996, Pfeiffer et al. 1998, Martini et al. 1998, Colls et al. 1998, Baumgartner
et al. 1999, Durak et al. 1999).
Acknowledgements. I am grateful to Dr. U. Trautmann and Dr. P. Binder-Wirth for sharing
their technical experience.