For reliability and reproducibility of results, we studied karyotypes of up to 10 roots and leaves from each individual plant and in each preparation we counted chromosomes in 5 to 10 metaphase plates
. Chromosome numbers were determined by using microscopes Biomed-2 (Biomed-Service Ltd., Moscow, Russia) and Axioskop-40 (Carl Zeiss AG, Oberkochen, Germany).
As the newly formed daughter nuclei pull apart and move away from the equatorial plate (metaphase plate
) region of the cell, small vesicles migrate to that area and associate with microtubule fibrils found there.
Oocyte was exposed to UV lights for localization of metaphase plate
and PB1 just before enucleation.
Therefore, higher rates of metaphase plate
and condensed chromatin in non-activated oocytes as compared to the activated ones in this study (Table 1) might be related to the lack of artificial induction of oocyte stimulation following ICSI in the former.
Digital images of well-spread metaphase plates
were captured using the Genus software (Applied Imaging Genetix, Queensway, UK).
The plane of the meiotic metaphase plate
of primary oocytes is parallel to the animal-vegetal axis.
The mean values and standard deviations of the relative lengths and total lengths of 18 chromosomes pairs were estimated from arm length measurements in well-spread unstained metaphase plates
from the larvae using chromosomal image analysis, the maximum length of the chromosomes was 5.67 [+ or -] 0.56 [micro]m and the minimum was 3.75 [+ or -] 0.071 [micro]m (Table 1).
Twenty-five newborns were included in each study group, but 27 newborns were excluded from the study because sufficient metaphase plates
could not be obtained and SCE frequency could not be studied.
Numerical abnormal metaphase plates
may contain more than 1 chromosome abnormality that each one by itself can be expressed and Ivisualized as MN in subsequent generating embryos.
Satisfactory results were obtained in 1 8 of the 25 specimens analysed and after counting 238 metaphase plates
a modal diploid number of 56 chromosomes were determined.
At least 50 well-spread metaphase plates
were counted and dividing cells in late prophase/early metaphase with about 450 band resolutions were karyotyped using the karyoimager software (Carl Zeiss, Germany).
Owing to the small number of suitable spermatogonial mitotic metaphase plates
, we used sister metaphase II for analysis of karyotypes in males.