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 Flow Cytometer is designed to deliver this precision regardless of the sample throughput rate. You no longer need to sacrifice throughput for sensitivity.

Learn more about the Attune® Cytometer

Download the cell cycle application note

Precision, Regardless of the Sample Throughput Rate

The figures below show the low-variability data obtained with the Attune® Acoustic Focusing Cytometer, even at high sample rates.

 Figure 3. Minimal data variation at high sample rates with the Attune® Acoustic Focusing Cytometer. Jurkat 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 rates 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/GRatio  %CV
High-End Hydrodynamic
Cytometer
12 µL/min41.7338.7320.441.96 4.83
35 µL/min40.1638.9520.891.96 6.12
60 µL/min44.6026.1729.231.90 7.76
Attune® Acoustic
Focusing Cytometer
25 µL/min45.2040.2914.511.94 3.22
100 µL/min42.8138.7218.741.94 3.17
1,000 µL/min40.2538.5521.201.94 4.21

More Cell Types

In another cell cycle study, five cell types (listed below) were analyzed with the Attune® cytometer and with 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