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Moreover, Figure 4c shows that SA β-Gal activity and large nuclei content were increased in aged hepatocytes, and Figure 4d shows the images for the increased SA β-Gal positive staining in aged hepatocytes that correlated with the increased p21 and p16 and diminished RAC3 expression detected by immunofluorescence performed in liver sections.
Frozen tissue was embedded in optimal cutting temperature compound and cryosections from the top and middle of specimens were stained with hematoxylin and eosin prior to being evaluated by a pathologist for the presence of >60% tumor nuclei content or absence of tumor (i.e., normal).
Both of these assays, in their commercial forms, have implemented quality control strategies to account for tumor nuclei content.
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Moreover, when cells from fresh tumors were dissociated and cultured overnight on glass coverslips, fused cells were observed by nuclei contents analysis by FISH (Figure 5B).
Although a significant elevation in CLI was found in aneuploid tumours this is likely to represent the systemic bias of the calculation methods, with no difference being seen between aneuploid and diploid tumours when BrdUrd labelling was measured with-out reference to the nuclei DNA content.
To quantitate DNA content, nuclei images of DAPI-stained animals were taken with a Zeiss AxioCam, and images were analyzed with NIH ImageJ 1.40 g software.
In addition, confocal microscopy revealed that some single cells had nuclei with abnormal content which may have resulted from a fusion event (nucleus with one Y and several X chromosomes) (data not shown).
Decellularized liver scaffolds processed from all SDS-based protocols are free of significant DNA content, nuclei, and cytoplasmic proteins.
In control experiments, AD cerebellar nuclei showed DNA content profiles similar to lymphocytes, characterized by histograms with sharp peaks and narrow bases.
Progression of meiosis was monitored by assessing the number of nuclei and DNA content per cell from fractions collected at 30-minute or 1-hour intervals during the synchronization procedure (Fig. 1b).
If a case classified as DNA diploid or DNA tetraploid demonstrated a 9c ER >0, this case was reclassified as DNA aneuploid, as the presence of tumour nuclei with DNA content exceeding 9c is believed to be evidence of aneuploidy (Motherby et al, 2002).
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