Error Prevention During Quality Control on the AQUIOS CL System

The AQUIOS CL system employs several features that work to prevent errors during quality control. These prevention methods can be broken down into the following:

  • Automatically Pause when QC Fails
    Refer to Laboratory SOP: Pause for QC in the Daily Quality Control on the AQUIOS CL System for the Tetra Application product bulletin for more about the Pause for QC function.
  • QC Checks
    Unlike traditional flow cytometers the AQUIOS CL flow cytometer does not require additional reagents such as fluorospheres to perform QC checks when running the Tetra application. The system’s advanced algorithm checks a variety of internal criteria and will simply display pass or fail. The AQUIOS relies on quantitative data to determine the pass/fail parameters. There are four categories of QC checks on the AQUIOS CL flow cytometer:
    • Daily QC (Controls)
      Daily QC is performed to ensure optical, fluidics and electronic stability within the system are operating within the instrument specifications. When enabled, the system provides a reminder to run controls. Refer to the Daily Quality Control on the AQUIOS CL System for the Tetra Application Product Bulletin for a detailed look at the QC process on the AQUIOS CL flow cytometer.
    • Real Time QC (Patient Sample)
      Real time QC for the Patient Sample automatically monitors several variables during a sample run including: separation quotient (SQ), mean channel, stable data rates, and compensation checks. Since there is no range established for a given sample, the system will not flag it. However, the system will display a generic notification if there are any inconsistencies.

Application-Specific Quality Checks (Patient Sample) 
AQUIOS Tetra Application. The AQUIOS Tetra application consists of three tests: the Tetra 1 test (CD45-FITC/CD4-RD1/CD8-ECD/CD3-PC5), the Tetra-2+ test (CD45-FITC/(CD56+CD16+) RD1/CD19- ECD/CD3-PC5), and the Tetra Combo test (AQUIOS Tetra 1 test followed by AQUIOS Tetra 2+ test). 
The AQUIOS Tetra 1 Panel reagent also functions as an analytic reliability check for a specimen by monitoring the Total CD3+ absolute count. The CD3+ Reliability Check is the sum of the percentages of CD3+CD4+ and CD3+CD8+ cells and should be within ±5% of the total percentage of CD3+. (5)

The combination of AQUIOS Tetra 2+ Panel monoclonal antibody reagents can function as a quality control check for a specimen in terms of total lymphocyte percentage determined using the following formula: (5)

FLOWC-HIV-aquios formula - Section 9

The combination of monoclonal antibody reagents in the Tetra Combo test can function as a quality control check for a specimen in terms of total lymphocyte percentage determined using the same formula described above.


CD3+ Intrapanel Check in the Tetra Combo test is the variability between AQUIOS Tetra 1 and AQUIOS Tetra 2+ for CD3+ and serves as an internal control. Differences between replicate CD3 percent positive results should be ±3.5%.

System Checks

The system automatically verifies the laser power and that the vacuum is acceptable before each sample is run. The system also verifies internal communications and that the hardware is functioning correctly at each step of the process. In addition, the system performs a fluidic check at the start of each run to ensure there is enough AQUIOS Sheath Solution available for the requested run.

With the AQUIOS CL flow cytometry system, QC checks associated with instrument performance essentially follow the same principles that are behind the fluorosphere-based QC checks performed by general purpose flow cytometers. The key difference is that the AQUIOS CL flow cytometer is designed to perform these QC checks automatically without the need for additional reagents. This helps to simplify the process making QC checks associated with the instrument quick and efficient.

AQUIOS CL Flow Cytometer References

1. Evaluation of a novel automated volumetric flow cytometer for absolute CD4+ T lymphocyte quantitation
Gossez M, Malcus C, Demaret J, Frater J, Poitevin-Later F, Monneret G
Cytometry B Clin Cytom. 2017 Nov;92(6):456-464. doi: 10.1002/cyto.b.21360. Epub 2016 Feb 23.
https://onlinelibrary.wiley.com/doi/full/10.1002/cyto.b.21360
2. Comparison of methods and TAT assessment: Volumetric AQUIOS CL and bead-based FACS CANTO II cytometers
Grossi V, Infantino M, Meacci F, Bellio E, Bellio V, Ciotta G, Priami F, Sarzi-Puttini P, Atzeni F, Li Gobbi F, Damiani A, Benucci M, Manfredi M
Cytometry B Clin Cytom. 2018 Jul;94(4):674-678. doi: 10.1002/cyto.b.21513. Epub 2017 Feb 15.
https://onlinelibrary.wiley.com/doi/abs/10.1002/cyto.b.21513
3. Performance verification of the new fully automated Aquios flow cytometer PanLeucogate (PLG) platform for CD4-T-lymphocyte enumeration in South Africa
Coetzee LM, Glencross DK
PLoS One. 2017; 12(11): e0187456. Published online 2017 Nov 3.  doi: 10.1371/journal.pone.0187456
https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0187456
4. T lymphocyte immunophenotyping in bronchoalveolar lavage on AQUIOS CL cytometer.
Humeau C, Darrouzain F, Gouilleux-Gruart V
J Immunol Methods. 2018 Oct;461:91-99. doi: 10.1016/j.jim.2018.06.022. Epub 2018 Jul 3.
https://www.sciencedirect.com/science/article/pii/S0022175918300668?via=ihub
5. Analytical performance of an automated volumetric flow cytometer for quantitation of T, B and natural killer lymphocytes.
Degandt S, Peeters B, Jughmans S, Boeckx N, Bossuyt X.
Clin Chem Lab Med. 2018 Jul 26;56(8):1277-1288. doi: 10.1515/cclm-2017-0638.
https://www.degruyter.com/view/j/cclm.2018.56.issue-8/cclm-2017-0638/cclm-2017-0638.xml
6. Acceptable "Real-Life" Variability for Lymphocyte Counts by Flow Cytometry
Ticchioni M, Brouzes C, Durrieu F, Lambert C; Association Française de Cytométrie accreditation working group (AFC-AG).
Cytometry B Clin Cytom. 2018 Dec 7. doi: 10.1002/cyto.b.21751. [Epub ahead of print]
https://onlinelibrary.wiley.com/doi/abs/10.1002/cyto.b.21751
7. Advantages of the Use of AQUIOS CL Flow Cytometer in Evaluation of CD4 and CD8 in HIV Patients: CMCVAMC Experience
Patel S, Wolf B, Jhala D
American Journal of Clinical Pathology, Volume 150, Issue suppl_1, 21 September 2018, Pages S70, https://doi.org/10.1093/ajcp/aqy093.170
https://academic.oup.com/ajcp/article/150/suppl_1/S70/5103090

Efficiently manage your high

volume routine applications

with AQUIOS CL,  which enables

automation of your most

routine, repetitive tasks.

FLOWC_HIV CD4 London health Sciences AQUIOS - Section 7

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