|
Experimental Approach In this experiment, we used the Cellomics iQ approach to collect 100 to 1,000 objects to investigate the effects of a targeted siRNA on the cell cycle of A549 cells. Polo-like kinase 1 (PLK1) is a check point enzyme which regulates G2M progression in mammalian cells1. - If less than 1,000 objects are acquired, the confidence (P) values are not statistically significant between the negative controls and experimental PLK1 group.
- Selecting a fixed number of fields will lead to poor assay results in wells that show response variability.
Figure 1. The number of objects collected greatly impacts statistical relevance. The number of fields required to obtain a certain number of objects in the G0/G1 (2N) phase of the cell cycle for both the negative controls as well as cells transfected with the siRNA PLK1. * = The top of each bar are the confidence (P) values for the percentage of cells in 2N for PLK1 versus negative controls. Statistical relevance is based on ANOVA with post hoc test. How can Cellomics iQ decrease time to decision? With Cellomics iQ, scientists get to the right number of cells sooner, plate after plate, increasing productivity and reproducibility of results reducing the risk of a failed screen. The diagram below describes the typical workflow using the Cellomics iQ method to screen the Human Genome (22,000 targeted genes) as compared to off-line or primitive on-line cell counting systems. 
Figure 2. The number of objects collected greatly impacts time to data and decision. This table outlines the design to accomplish entire siRNA genome screens of mouse or human origin on 96-well plates. *Calculations assume 40 siRNAs run in duplicate on each 96-well plate plus appropriate controls and 16 minutes or less per plate to obtain 1,000 imaged and analyzed objects per well; number of days is based on imaging 24 hours per day; siRNA library sizes were obtained from www.dharmacon.com 
iQ leads to quicker, cheaper and more efficient decision making. 1Ree, AH, et al. (March-April 2004) "Ionizing Radiation Inhibits the PLK Cell Cycle Gene in a G2 Checkpoint-Dependent Manner." Anticancer Research. 24(2B): 555-562. Cellomics is a registered trademark of Cellomics, Inc.
© 2007 Thermo Fisher Scientific Inc. All rights reserved.
Cell Cycle – LC01571300 |
Home 
How many cells is enough? Targeting the right number of cells.
top