Cells were fixed by adding 4% paraformaldehyde in PBS directly into the culture medium at a 1:1 para:medium ratio and incubated for 15 min at RT. Cells were then washed 3× in PBS and stained with DRAQ5 (Biostatus Limited) 1:5,000 in PBS. Automated imaging steps were performed using an Opera system (Perkin Elmer). Images were taken using a 488/640 nm excitation laser (1st acquisition) and a 568 nm excitation laser (2nd acquisition). Images were analyzed using the Acapella software package (Perkin-Elmer). The Green/Red ratio was calculated as the ratio between the average nuclear intensity signal in the 488 nm channel and the average nuclear signal in the 568 nm channel. Minimum of a 1000 nuclei were analyzed in each experiment condition.
Acapella software package
Manufactured by PerkinElmer
Acapella is a software package developed by PerkinElmer to analyze and process high-content screening data. It provides tools for image segmentation, object identification, and feature extraction from microscopy images.
Automatically generated - may contain errors
Lab products found in correlation
3 protocols using acapella software package
1
Cell Fixation and Fluorescence Imaging
2
Cell Fixation and Fluorescence Imaging
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Cells were fixed by adding 4% paraformaldehyde in PBS directly into the culture medium at a 1:1 para:medium ratio and incubated for 15 min at RT. Cells were then washed 3× in PBS and stained with DRAQ5 (Biostatus Limited) 1:5,000 in PBS. Automated imaging steps were performed using an Opera system (Perkin Elmer). Images were taken using a 488/640 nm excitation laser (1st acquisition) and a 568 nm excitation laser (2nd acquisition). Images were analyzed using the Acapella software package (Perkin-Elmer). The Green/Red ratio was calculated as the ratio between the average nuclear intensity signal in the 488 nm channel and the average nuclear signal in the 568 nm channel. Minimum of a 1000 nuclei were analyzed in each experiment condition.
3
Quantitative Fluorescence Imaging Protocol
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Cells were fixed by adding 4% paraformaldehyde in phosphate buffered saline (PBS) directly into the culture medium and incubated for 15 min at RT. Cells were then washed 3× in PBS and stained with DRAQ5 (Biostatus Limited) 1:5, 000 in PBS. The automated imaging steps were performed using an Opera system (Perkin Elmer). Images were taken using a 488/640 nm excitation laser (1st acquisition) and a 568 nm excitation laser (2nd acquisition). Images were analyzed using the Acapella software package (Perkin-Elmer). The Green/Red ratio was calculated as the ratio between the average nuclear intensity signal in the 488 nm channel and the average nuclear signal in the 568 nm channel.
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