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  7. Cell Cycle Analysis With The Attune® Flow Cytometer

Cell Cycle Analysis With The Attune® Flow Cytometer

Cell Cycle Analysis With The Attune® Flow Cytometer

Accurate cell cycle phase assignments rely on precise measurements of fluorescence intensity differences between multiple cell populations. The Attune® Acoustic Focusing Cytometer delivers this precision regardless of the sample throughput rate. You no longer need to sacrifice throughput for sensitivity.

Precision, Regardless the Sample Throughput Rate


  Figure 3. Minimal data variation at high sample rates with the Attune® Acoustic Focusing Cytometer. Jurat cells were fixed and stained with propidium iodide, treated with RNase, and analyzed at a concentration of 1 x 104 cells/mL on a high-end instrument that uses hydrodynamic focusing, and on the Attune® Acoustic Focusing Cytometer at different sample rates. The left peak in all graphcs reflects cells in G0/G1 phase, while the right peak reflects cells in G2/M phase. Note that as sample rates increase on the instrument that uses hydrodynamic focusing, the width of the G0/G1 and G2/M peaks increase, whereas for the Attune® cytometer the peaks are relatively stable, even at the highest sample rate of 1,000 µL/min. These results are quantitated in Table 1.

 
Figure 4. Percent CV of G0/G1 at different flow rate on the Attune® cytometer and a competitor (C) at different smaple rates. Note the minimal change in variability (%CV) for the Attune® Acoustic Focusing Cytometer, even at a high sample rate.

Table 1.


  Sample Rate
 %G0/G1
 %S* %G2/M G0/G1 Ratio %CV
High-End Hydrodynamic Cytometer 12 µL/min 41.73 38.73 20.44 1.96 4.83
35 µL/min 40.16 38.95 20.89 1.96 6.12
60 µL/min 44.60 26.17 29.23
 1.90 7.76
Attune® Acoustic Focusing Cytometer 25 µL/min 45.20 40.29 14.51 1.94 3.22
100 µL/min 42.81 38.72 18.74 1.94 3.17
1,000 µL/min 40.25 38.55 21.20 1.94 4.21

More Cell Types


In another cell cycle study, five cell types (listed below) were analyzed in the Attune® cytometer and in a high-end hydrodynamic focusing cytometer (ModFit analysis was performed in each case). The sample throughput rate of 25 µL/min was used for the analysis.

Additional cell types analyzed with the Attune® Flow Cytometer:

  • Ramos B CellsKG-1
  • KG-1 a Myeloblast Cells
  • HL60 Promyeloblast Cells
  • U266
  • Myeloma  Cells
  • ST486 B Cells

 



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