The AQUIOS CL Flow Cytometer is intended for use with in vitro diagnostic flow cytometric applications using up to four fluorescent detection channels using a blue (488 mm) laser, two light scatter detection volume (EV). It is used in conjunction with the following reagents and software package.

AQUIOS Tetra-1 Panel and AQUIOS Tetra-2+ Panel monoclonal antibody reagents are for use on the AQUIOS CL Flow Cytometer with peripheral whole blood for immunophenotyping. These reagents are indicated for use in the immunologic assessment of patients having, or suspected of having, immune deficiency. These reagents provide identification and enumeration of;

AQUIOS Tetra-1 Panel Monoclonal Antibody Reagent

· Total CD3+, CD3+CD4+,CD3+CD3+, CD3+CD4+/CD3+CD8+ (ratio only) lymphocyte percentages and absolute counts.

• CD45+ absolute count

· CD45+ Low SS (lymphocytes) percentage and absolute count
AQUIOS Tetra-2+ Panel Monoclonal Antibody Reagent
· Total CD3+, CD3-CD19+, CD3-CD56+ and/or CD16+ lymphocyte percentages and absolute counts.
CD45+ absolute count
· CD45+ Low SS (lymphocytes) percentage and absolute count

AQUIOS Flow Cytometry Software may be run on an independent computer workstation for off-line analysis of results generated by the AQUIOS CL Flow Cytometer with the monoclonal antibody reagents listed above. The off-line analysis must be performed in accordance with the product labeling.

AQUIOS Tetra-1 Panel Monoclonal Antibody Reagent

· Total CD3+, CD3+CD4+,CD3+CD8+, CD3+CD4+/CD3+CD8+(ratio only) lymphocyte percentages and absolute

counts.

CD45+ absolute count
· CD45+ Low SS (lymphocytes) percentage and absolute count
AQUIOS Tetra-2+ Panel Monoclonal Antibody Reagent
· Total CD3+, CD3-CD19+, CD3-CD56+ and/or CD16+ lymphocyte percentages and absolute counts.
CD45+ absolute count
· CD45+ Low SS (lymphocytes) percentage and absolute count

AQUIOS IMMUNO-TROL Cells are assayed, lysable whole blood quality control product for immunophenotyping analysis using monoclonal antibody reagents and flow cytometry. It provides a positive cell control that is processed in the same manner as a whole blood sample. This allows verification of instrument and reagent performance. It also verifies the methods used for staining targeted cells, lysing erythrocytes, and analyzing samples by the AQUIOS CL Flow Cytometer.

AQUIOS IMMUNO-TROL Low Cells are assayed, lysable whole blood quality control product for immunophenotyping analysis using monoclonal antibody reagents and flow cytometry. It provides a positive cell control that is processed in the same manner as a whole blood sample. This allows verification of instrument and reagent performance. It also verifies the methods used for staining targeted cells, lysing erythrocytes, and analyzing samples by the AQUIOS CL Flow Cytometer.

AQUIOS Lysing Reagent Kit is used as part of the AQUIOS flow cytometer system. The kit consists used by AQUIOS flow cytometers to prepare whole blood samples for analysis of white blood cells.

Device Description

The AQUIOS CL Flow Cytometry System is composed of the following components:

. AQUIOS CL Flow Cytometer
AQUIOS System Software ●
AQUIOS Tetra-1 Panel CD45-FITC/CD4-RD1/CD8-ECD/CD3-PC5 ●
AQUIOS Tetra-2+ Panel CD45-FITC/(CD56+CD16)-RD1/CD19-ECD/CD3-PC5 ●
AQUIOS Immuno-Trol Cells ●
AQUIOS Immuno-Trol Low Cells ●
. AQUIOS Lysing Reagent Kit

The AQUIOS CL Flow Cytometer uses flow cytometric principles to determine qualitative and quantitative measurements of biological and physical properties of cells and other particles. These properties are measured when the cells pass through the laser beam(s) in single file.

The AQUIOS System Software is designed for the AQUIOS CL flow cytometer. It includes the algorithms and test definitions that provide automated analysis and results for AQUIOS Tetra-1 and 2+ reagents; this application cannot be modified by the user.

The AQUIOS Flow Cytometry System also offers an optional standalone offline workstation. This workstation is identical to the workstation that is physically connected to the instrument and can be used for off-line analysis of results generated by the AQUIOS CL Flow Cytometer with AQUIOS Tetra-1 and Tetra-2+ reagents and AQUIOS System software according to the product labeling.

AQUIOS Tetra-1 Panel CD45-FITC/CD4-RD1/CD8-ECD/CD3-PC5 reagent provides identification and enumeration of CD45+, CD45+ Low SS, and CD3+/CD4+, CD3+/CD8+, and CD3+ lymphocyte percentages and absolute counts in peripheral whole blood. AQUIOS Tetra-2+ Panel CD45-FITC/(CD56+CD16)-RD1/CD19-ECD/CD3-PC5 provides identification and enumeration of CD45+, CD45+ Low SS, and CD3+, CD3-/CD19+ and CD3-/CD56+CD16+ lymphocyte percentages and absolute counts in peripheral whole blood. Additionally, both panels provide for CD45+ absolute count and CD45+ Low SS absolute count and percentage.

AQUIOS Immuno-Trol and Immuno-Trol Low Cells are assayed, lysable whole blood quality control product for immunophenotyping analysis using monoclonal antibody reagents and flow cytometry. It provides a positive cell control that is processed in the same manner as a whole blood sample. This allows verification of instrument and reagent performance. It also verifies the methods used for staining targeted cells, lysing erythrocytes, and analyzing samples by the AQUIOS CL Flow Cytometer.

The AQUIOS CL Flow Cytometer uses on-board sample preparation as part of the overall system workflow. The AQUIOS Lysing Reagent Kit is comprised of two readyto-use reagents: Reagent A lyses the red blood cells, Reagent B quenches the solution, slowing the lyse reaction down in preparation for analysis. This reagent system provides a rapid, no-wash, standardized, whole blood lysing solution for sample to sample, and laboratory to laboratory reproducibility.

AI/ML Overview

Here's an analysis of the acceptance criteria and study findings for the AQUIOS CL Flow Cytometry System, based on the provided document:

1. Table of Acceptance Criteria and Reported Device Performance

The document provides summary results rather than explicit, quantified acceptance criteria for many tests. However, it consistently states that the device "meets performance requirements" or "was demonstrated." Where specific performance measures or outcomes are mentioned, they are included below.

Study Category	Specific Study/Characteristic	Acceptance Criteria (Implied/Stated)	Reported Device Performance
Instrument Performance	Fluorescence Linearity	Linearity of fluorescence measurements (implied: within acceptable limits)	Linearity of fluorescence measurements was demonstrated.
	Electronic Volume Linearity	Linearity of Electronic Volume measurements (implied: within acceptable limits)	Linearity of Electronic Volume measurements was demonstrated.
	Laser Performance Characteristics	Stability of the laser performance over time (implied: within acceptable limits)	The AQUIOS CL laser performance is stable over time.
	Analyzer Carryover (Whole Blood)	Meet performance specifications for carryover	Meets the whole blood carryover performance requirements.
	Instrument Settings Stability	Stability of instrument settings to support adequacy of QC methodology	Stability of the AQUIOS CL instrument settings support the quality control methodology.
	Gravimetrics (Dispensing Accuracy/Precision)	Accuracy and precision of dispensing lysing/antibody reagents and specimen aspiration/dispense	Accuracy and precision of dispensing AQUIOS lysing and antibody reagents, and specimen aspiration/dispense are demonstrated.
Reagent/Assay Performance	Assay Linearity (Lymphocyte Subsets)	Linear range of absolute values for each lymphocyte subset population (implied: within acceptable limits)	Meets the linearity performance requirements.
	Assay Carryover (Whole Blood and Reagents)	Meet performance specifications for carryover	Meets carryover performance requirements.
	Detection Capability (LoB, LLoD, LLoQ)	Meet performance requirements for LoB, LLoD, and LLoQ in whole blood	Meets the performance requirements for LoB, LLoD, and LLoQ in whole blood.
	Specimen and Prepared Sample Stability	Verify specimen and prepared sample stability claims (implied: within specified time/conditions)	Meets the requirements for specimen and prepared sample stability.
	Method Comparison (vs. Predicate)	Bias between subject device and predicate (implied: within acceptable limits)	Meets the performance requirements when compared to the predicate device.
	Precision - Long Term Imprecision	Demonstrates system imprecision (implied: within acceptable limits)	Meets performance requirements for Long Term Imprecision.
	Precision - Whole Blood Repeatability	Estimate repeatability at various medical decision levels and data percentiles (implied: within acceptable limits)	Meets performance requirements for Whole Blood Repeatability.
	Comparability (Anticoagulant, Sample Mode, Test Panel)	Equivalency across anticoagulants, modes, and panels	Within-method comparisons are equivalent.
	Adult Reference Intervals	Establish intervals consistent with published values for T, B, and NK lymphocyte subsets	Established and confirmed consistent with published values.
Reagent Stability/Variability	AQUIOS Tetra-1 & Tetra-2+ Reagent Stability	Demonstrate shelf life (implied: meets specified duration and conditions)	Meets performance requirements in support of stability claims.
	AQUIOS Tetra-1 & Tetra-2+ Lot Variability	Acceptable lot variability of material	Has acceptable lot variability performance.
	AQUIOS Immuno-Trol & Low Cells Assay Value Assignment/Ranges	Determine and verify expected ranges/process for assignment	Assay value assignment process and expected ranges were established and verified.
	AQUIOS Immuno-Trol & Low Cells Stability	Demonstrate shelf life (implied: meets specified duration and conditions)	Meets performance requirements in support of stability claims.
	AQUIOS Immuno-Trol & Low Cells Lot Variability	Acceptable lot variability of material	Has acceptable lot variability performance.
	AQUIOS Lysing Reagent Kit Stability	Demonstrate shelf life (implied: meets specified duration and conditions)	Meets performance requirements in support of stability claims.
	AQUIOS Lysing Reagent Kit Lot Variability	Acceptable lot variability of material	Has acceptable lot variability performance.

2. Sample Sizes Used for the Test Set and Data Provenance

The document does not explicitly state the sample sizes used for the test set portion of the studies (which would typically involve independent validation after development). Instead, it describes general testing approaches. It also does not specify the country of origin for the data or whether it was retrospective or prospective.

For example, for "Method Comparison," it merely states "To evaluate bias between the subject device versus the predicate" and uses CLSI EP09-A3 (Method Comparison and Bias Estimation Using Patient Samples), which implies patient samples were used, but no quantity or demographics are given. Similarly, "Adult Reference Intervals" implies a study on adults, but sample size and demographics are not specified.

3. Number of Experts Used to Establish Ground Truth for the Test Set and Qualifications

The document does not mention the use of experts to establish a "ground truth" for a test set in the traditional sense of image interpretation or complex diagnostic decision-making. The AQUIOS CL Flow Cytometry System is an automated cell counter. Its "ground truth" for the test set would typically be established by established reference methods, predicate devices, or validated laboratory assays (e.g., manual differential counts, confirmed cell populations by expert flow cytometrists, or validated internal methods). The document refers to "predicate devices" for comparison, which themselves are legally marketed and validated, serving as a de facto "ground truth" in terms of established performance.

4. Adjudication Method for the Test Set

Not applicable. As "ground truth" is not explicitly established by human experts in a subjective interpretation process (like in radiology), an adjudication method in the sense of resolving inter-reader disagreements is not described or implied. The system relies on comparisons to predicate devices and validated methods.

5. If a Multi-Reader Multi-Case (MRMC) Comparative Effectiveness Study Was Done

No. An MRMC comparative effectiveness study, which typically compares human reader performance with and without AI assistance, was not conducted. This device is an automated in vitro diagnostic system for cell counting and immunophenotyping, not a system that assists human readers in interpreting complex cases. Its "effectiveness" is measured by its analytical performance (accuracy, precision, linearity, etc.) against established laboratory methods and predicate devices.

6. If a Standalone (Algorithm Only Without Human-in-the-Loop) Performance Was Done

Yes, implicitly. The entire set of performance studies described (Fluorescence Linearity, Electronic Volume Linearity, Laser Performance, Analyzer Carryover, Instrument Settings Stability, Gravimetrics, Comparability, Assay Linearity, Assay Carryover, Detection Capability, Specimen and Prepared Sample Stability, Method Comparison, Precision, Adult Reference Intervals, Reagent Stability, Lot Variability) evaluates the device's autonomous performance. The AQUIOS CL Flow Cytometer, along with its reagents and software, is designed to perform these tasks automatically. While human operators are involved in sample preparation and loading, the core measurements and classifications are driven by the instrument's algorithms and hardware, making these standalone performance assessments. The "AQUIOS System Software" also includes algorithms for automated analysis.

7. The Type of Ground Truth Used

The ground truth for the performance studies appears to be based on:

Comparison to predicate devices: For analytical accuracy (e.g., "Method Comparison" against FACSCalibur or UniCel DxH 800). This means the established, cleared performance of the predicate serves as the standard.
Validated internal methods/specifications: For various instrument and assay characteristics like linearity, precision, detection limits, and stability.
Published values: For establishing "Adult Reference Intervals," which were confirmed to be "consistent with published values for T, B, and NK lymphocyte subsets."
CLSI (Clinical and Laboratory Standards Institute) guidelines and standards: These provide the methodological framework and often implicit performance thresholds for many tests (e.g., CLSI EP06-A for linearity, CLSI EP5-A2 for precision, CLSI H26-A2 for hematology analyzers).

8. The Sample Size for the Training Set

The document does not specify a separate "training set" or its size. For an IVD device like this, which performs quantitative measurements based on biophysical properties, the development process generally involves extensive internal testing and refinement (calibration, optimization, verification) using a wide range of samples, rather than a distinct "training set" in the machine learning sense. The performance studies described in the document are primarily for validation and verification.

9. How the Ground Truth for the Training Set Was Established

Since a "training set" in the AI/ML context is not explicitly mentioned, the concept of establishing ground truth for it is not directly addressed. Instead, the analytical methods (flow cytometry principles, reagent chemistries, software algorithms) are based on well-established scientific principles and calibrated against reference materials and methods during the device's development and manufacturing.

Summary

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Image /page/0/Picture/1 description: The image shows the seal of the Department of Health & Human Services (HHS) of the United States of America. The seal features the department's name encircling a stylized eagle emblem. The eagle is depicted in profile, with its head facing right, and its feathers are represented by a series of curved lines.

Food and Drug Administration 10903 New Hampshire Avenue Document Control Center - WO66-G609 Silver Spring, MD 20993-0002

January 27, 2016

Beckman Coulter, Inc. c/o Mr. Anthony Dennis Staff Regulatory Affairs Specialist 11800 SW 147th Avenue, M/S 31-B06 Miami, FL 33196-2500

Re: K141932

Trade/Device Name: AQUIOS CL Flow Cytometry System AQUIOS Tetra-1 and Tetra-2+ Monoclonal Antibody Reagents AQUIOS Lysing Reagent Kit AQUIOS Immuno-Trol and Immuno-Trol Low Cells Regulation Number: 21 CFR §864.5220 Regulation Name: Automated differential cell counter Regulatory Class: Class II Product Code: OYE, JPK, GGK Dated: March 5, 2015 Received: March 9, 2015

Dear Mr. Dennis:

This letter corrects our substantially equivalent letter of April 10, 2015.

We have reviewed your Section 510(k) premarket notification of intent to market the device referenced above and have determined the device is substantially equivalent (for the indications for use stated in the enclosure) to legally marketed predicate devices marketed in interstate commerce prior to May 28, 1976, the enactment date of the Medical Device Amendments or to devices that have been reclassified in accordance with the provisions of the Federal Food, Drug, and Cosmetic Act (Act) that do not require approval of a premarket approval application (PMA). You may, therefore, market the device, subject to the general controls provisions of the Act. The general controls provisions of the Act include requirements for annual registration, listing of devices, good manufacturing practice, labeling, and prohibitions against misbranding and adulteration. Please note: CDRH does not evaluate information related to contract liability warranties. We remind you, however, that device labeling must be truthful and not misleading.

If your device is classified (see above) into either class II (Special Controls) or class III (PMA), it may be subject to additional controls. Existing major regulations affecting your device can be found in the Code of Federal Regulations, Title 21, Parts 800 to 898. In addition, FDA may publish further announcements concerning your device in the Federal Register.

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Please be advised that FDA's issuance of a substantial equivalence determination does not mean that FDA has made a determination that your device complies with other requirements of the Act or any Federal statutes and regulations administered by other Federal agencies. You must comply with all the Act's requirements, including, but not limited to: registration and listing (21 CFR Part 807); labeling (21 CFR Parts 801 and 809] ); medical device reporting (reporting of medical device-related adverse events) (21 CFR 803); good manufacturing practice requirements as set forth in the quality systems (OS) regulation (21 CFR Part 820); and if applicable, the electronic product radiation control provisions (Sections 531-542 of the Act); 21 CFR 1000-1050.

If you desire specific advice for your device on our labeling regulation (21 CFR Parts 801 and 809]), please contact the Division of Industry and Consumer Education at its toll-free number (800) 638-2041 or (301) 796-7100 or at its Internet address http://www.fda.gov/MedicalDevices/Resourcesfor You/Industry/default.htm. Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21CFR Part 807.97). For questions regarding the reporting of adverse events under the MDR regulation (21 CFR Part 803), please go to http://www.fda.gov/MedicalDevices/Safety/ReportaProblem/default.htm for the CDRH's Office of Surveillance and Biometrics/Division of Postmarket Surveillance.

You may obtain other general information on your responsibilities under the Act from the Division of Industry and Consumer Education at its toll-free number (800) 638-2041 or (301) 796-7100 or at its Internet address

http://www.fda.gov/MedicalDevices/ResourcesforYou/Industry/default.htm.

Sincerely yours,

Leonthena R. Carrington -S

Leonthena R. Carrington, MS, MBA, MT(ASCP) Director Division of Immunology and Hematology Devices Office of In Vitro Diagnostics and Radiological Health Center for Devices and Radiological Health

Enclosure

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510(k) Number (if known) K141932

Device Name AQUIOS CL Flow Cytometry System

Indications for Use (Describe)

AQUIOS Tetra-1 Panel Monoclonal Antibody Reagent

· Total CD3+, CD3+CD4+,CD3+CD3+, CD3+CD4+/CD3+CD8+ (ratio only) lymphocyte percentages and absolute counts.

• CD45+ absolute count

· CD45+ Low SS (lymphocytes) percentage and absolute count
AQUIOS Tetra-2+ Panel Monoclonal Antibody Reagent
· Total CD3+, CD3-CD19+, CD3-CD56+ and/or CD16+ lymphocyte percentages and absolute counts.
CD45+ absolute count
· CD45+ Low SS (lymphocytes) percentage and absolute count

Type of Use (Select one or both, as applicable)

Prescription Use (Part 21 CFR 801 Subpart D)
Over-The-Counter Use (21 CFR 801 Subpart C)

CONTINUE ON A SEPARATE PAGE IF NEEDED.

This section applies only to requirements of the Paperwork Reduction Act of 1995.

DO NOT SEND YOUR COMPLETED FORM TO THE PRA STAFF EMAIL ADDRESS BELOW.

The burden time for this collection of information is estimated to average 79 hours per response, including the time to review instructions, search existing data sources, gather and maintain the data needed and complete and review the collection of information. Send comments regarding this burden estimate or any other aspect of this information collection, including suggestions for reducing this burden, to:

Department of Health and Human Services Food and Drug Administration Office of Chief Information Officer Paperwork Reduction Act (PRA) Staff PRAStaff@fda.hhs.gov

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510(k) Number (if known) K141932

Device Name

AQUIOS Tetra-1 and Tetra-2+ Monoclonal Antibody Reagents

Indications for Use (Describe)

AQUIOS Tetra-1 Panel Monoclonal Antibody Reagent

· Total CD3+, CD3+CD4+,CD3+CD8+, CD3+CD4+/CD3+CD8+(ratio only) lymphocyte percentages and absolute

counts.

CD45+ absolute count
· CD45+ Low SS (lymphocytes) percentage and absolute count
AQUIOS Tetra-2+ Panel Monoclonal Antibody Reagent
· Total CD3+, CD3-CD19+, CD3-CD56+ and/or CD16+ lymphocyte percentages and absolute counts.
CD45+ absolute count
· CD45+ Low SS (lymphocytes) percentage and absolute count

Type of Use (Select one or both, as applicable)
-------------------------------------------------

|X Prescription Use (Part 21 CFR 801 Subpart D)

| | Over-The-Counter Use (21 CFR 801 Subpart C)

CONTINUE ON A SEPARATE PAGE IF NEEDED.

This section applies only to requirements of the Paperwork Reduction Act of 1995.

DO NOT SEND YOUR COMPLETED FORM TO THE PRA STAFF EMAIL ADDRESS BELOW.

Department of Health and Human Services Food and Drug Administration Office of Chief Information Officer Paperwork Reduction Act (PRA) Staff PRAStaff@fda.hhs.gov

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510(k) Number (if known) K141932

Device Name AQUIOS Lysing Reagent Kit

Indications for Use (Describe)

AQUIOS Lysing Reagent Kit is used as part of the AQUIOS flow cytometer system. The kit consists used by AQUIOS flow cytometers to prepare whole blood samples for analysis of white blood cells. Type of

Type of Use (Select one or both, as applicable)
× Prescription Use (Part 21 CFR 801 Subpart D)	Over-The-Counter Use (21 CFR 801 Subpart C)

CONTINUE ON A SEPARATE PAGE IF NEEDED.

This section applies only to requirements of the Paperwork Reduction Act of 1995.

DO NOT SEND YOUR COMPLETED FORM TO THE PRA STAFF EMAIL ADDRESS BELOW.

Department of Health and Human Services Food and Drug Administration Office of Chief Information Officer Paperwork Reduction Act (PRA) Staff PRAStaff@fda.hhs.gov

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510(k) Number (if known) K141932

Device Name

AQUIOS Immuno-Trol and Immuno-Trol Low Cells

Indications for Use (Describe)

Type of Use (Select one or both, as applicable)

☒ Prescription Use (Part 21 CFR 801 Subpart D)
☐ Over-The-Counter Use (21 CFR 801 Subpart C)

CONTINUE ON A SEPARATE PAGE IF NEEDED.

This section applies only to requirements of the Paperwork Reduction Act of 1995.

DO NOT SEND YOUR COMPLETED FORM TO THE PRA STAFF EMAIL ADDRESS BELOW.

Department of Health and Human Services Food and Drug Administration Office of Chief Information Officer Paperwork Reduction Act (PRA) Staff PRAStaff@fda.hhs.gov

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510(k) Summary for AQUIOS CL Flow Cytometry System

510(k) Owner / Submitter Information

Name: Beckman Coulter Inc. Address: 11800 SW 147th Ave., Miami, FL 33196 Phone #: (305) 380-4509 Fax #: (305) 380-4344 Contact Person: Anthony Dennis Email Address: adennis@beckman.com Date Updated: April 9, 2015

Device Information

Trade Name: AOUIOS CL Flow Cytometry System Common Name: AQUIOS CL Classification Name: Automated differential cell counter (21 CFR 864.5220) Classification: Class II Product Code: OYE Panel: Hematology

Trade Name: AQUIOS Tetra-1 and Tetra-2+ Monoclonal Antibody Reagents Common Name: AQUIOS Tetra-1 Panel CD45-FITC/CD4-RD1/CD8-ECD/CD3-PC5 and AQUIOS Tetra-2+ Panel CD45-FITC/(CD56+CD16)-RD1/CD19-ECD/CD3-PC5 Classification Name: Automated differential cell counter (21 CFR 864.5220) Classification: Class II Product Code: OYE Panel: Hematology

Trade Name: AOUIOS Lysing Reagent Kit

Common Name: AQUIOS Lysing Reagent Kit Classification Name: Red cell lysing reagent (21 CFR 864.8540), Automated differential cell counter (21 CFR 864.5220) Classification: Class I Non-Exempt [due to limitations of the exemptions of 21 CFR 864.9(c)(1) and (c)(3)] Product Code: GGK, OYE [as used as part of the AQUIOS flow cytometry system] Panel: Hematology

Trade Name: AOUIOS Immuno-Trol and Immuno-Trol Low Cells Common Name: AOUIOS Immuno-Trol and Immuno-Trol Low Classification Name: Hematology quality control mixture (21 CFR 864.8625) Classification: Class II Product Code: JPK Panel: Hematology

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Predicate Device Information

Predicate Product	510(k)Number	DateCleared	Classification	21 CFR	ProductCode
CaliBRITE 3 Color andFACSComp	K961623	6/07/96	Class II	864.5220	GKZ
CaliBRITE APC beads CaliBRITE4 kit (unlabeled, FITC-, PE-,PerCP- and APC- labeledCaliBRITE beads), andFACSComp software(FACSCalibur)	K973483	2/17/98	Class II	864.5220	GKZ
MultiTEST CD3/CD8/CD45/CD4	K974360	03/11/98	Class II	864.5220	GKZ
MultitestCD3/CD16+56/CD45/CD19Reagent and Multitest IMK KitLysing Solution with BD TruCountTubes	K980858	5/22/98	Class II	864.5220	GKZ
Unicel DxH 800	K140911	09/05/14	Class II	864.5220	GKZ
Immuno-Trol Control Cells	K984216	03/16/99	Class II	864.8625	JPK
Immuno-Trol Low Cells	K013842	12/13/01	Class II	864.8625	JPK

Device Description

The AQUIOS CL Flow Cytometry System is composed of the following components:

. AQUIOS CL Flow Cytometer
AQUIOS System Software ●
AQUIOS Tetra-1 Panel CD45-FITC/CD4-RD1/CD8-ECD/CD3-PC5 ●
AQUIOS Tetra-2+ Panel CD45-FITC/(CD56+CD16)-RD1/CD19-ECD/CD3-PC5 ●
AQUIOS Immuno-Trol Cells ●
AQUIOS Immuno-Trol Low Cells ●
. AQUIOS Lysing Reagent Kit

The AQUIOS CL Flow Cytometer uses flow cytometric principles to determine qualitative and quantitative measurements of biological and physical properties of cells

{8}------------------------------------------------

and other particles. These properties are measured when the cells pass through the laser beam(s) in single file.

Intended Use:

AQUIOS CL Flow Cytometry System Indications for Use:

The AQUIOS CL Flow Cytometer is intended for use with in vitro diagnostic flow cytometric applications using up to four fluorescent detection channels using a blue (488 nm) laser, two light scatter detection channels and electronic volume (EV). It is used in conjunction with the following reagents and software package.

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AQUIOS Tetra-1 Panel and AQUIOS Tetra-2+ Panel monoclonal antibody reagents are for use on the AQUIOS CL Flow Cytometer with peripheral whole blood for immunophenotyping. These reagents are indicated for use in the immunologic assessment of patients having, or suspected of having, immune deficiencv. These reagents provide identification and enumeration of;

AQUIOS Tetra-1 Panel Monoclonal Antibody Reagent -
- · Total CD3+, CD3+CD4+,CD3+CD8+, CD3+CD4+/CD3+CD8+ (ratio only) lymphocyte percentages and absolute counts.
- CD45+ absolute count
- CD45+ Low SS (lymphocytes) percentage and absolute count
AQUIOS Tetra-2+ Panel Monoclonal Antibody Reagent –
- · Total CD3+, CD3-CD19+, CD3-CD56+ and/or CD16+ lymphocyte percentages and absolute counts.
- CD45+ absolute count
- · CD45+ Low SS (lymphocytes) percentage and absolute count

AQUIOS Tetra-1 and Tetra-2 Indications for Use:

AQUIOS Tetra-1 Panel Monoclonal Antibody Reagent -
- · Total CD3+, CD3+CD4+,CD3+CD8+, CD3+CD4+/CD3+CD8+ (ratio only) lymphocyte percentages and absolute counts.
- CD45+ absolute count
- CD45+ Low SS (lymphocytes) percentage and absolute count
AQUIOS Tetra-2+ Panel Monoclonal Antibody Reagent -
- · Total CD3+, CD3-CD19+, CD3-CD56+ and/or CD16+ lymphocyte percentages and absolute counts.
- CD45+ absolute count
- · CD45+ Low SS (lymphocytes) percentage and absolute count

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AQUIOS Immuno-Trol and Immuno-Trol Low Cells Indications for Use:

AQUIOS Lysing Reagent Kit:

AQUIOS Lysing Reagent Kit is used as part of the AQUIOS flow cytometer system. The kit consists of two reagents used by AQUIOS flow cytometers to prepare whole blood samples for analysis of white blood cells.

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Technological Characteristics Comparisons to Predicate

AQUIOS CL Flow Cytometer Device Comparison Tables:

Similarities
	Predicate		Device
		FACSCalibur
Characteristic	CaliBRITE APC beads CaliBRITE 4kit (unlabeled, FITC-, PE-, PE-, PerCP-and APC- labeled CaliBRITE beads),and FACSComp software	CaliBRITE 3 Color and FACSComp	AQUIOS CL Flow Cytometer
Intended Use*	For flow cytometer set up and	For flow cytometer set up and	The AQUIOS CL Flow Cytometer is
	monitoring of instrument	monitoring of instrument	intended for use with in vitro diagnostic
	performance prior to performing	performance.	flow cytometric applications using up to
	reticulocyte enumeration or		four fluorescent detection channels using
	immunophenotyping applications.		a blue (488 nm) laser, two light scatter
	Flow cytometry has been found		detection channels and electronic
	useful in monitoring some forms of		volume (EV). It is used in conjunction
	immune disease.		with the following reagents and software
			package.
			AQUIOS Tetra-1 Panel and AQUIOS
			Tetra-2+ Panel monoclonal antibody
			reagents are for use on the AQUIOS CL
			Flow Cytometer with peripheral whole
	* Only similar parameters of both		blood for immunophenotyping. These
	devices are presented here. Refer to		reagents are indicated for use in the
	DxH 800 SE table for other		immunologic assessment of patients
	parameters.		having, or suspected of having, immune
			deficiency. These reagents provide
Similarities
Characteristic	PredicateFACSCalibur	Device
	CaliBRITE 3 Color and FACSComp	AQUIOS CL Flow Cytometer
		identification and enumeration of;
CaliBRITE APC beads CaliBRITE 4 kit (unlabeled, FITC-, PE-, PerCP- and APC- labeled CaliBRITE beads), and FACSComp software		- AQUIOS Tetra-1 Panel Monoclonal Antibody Reagent Total CD3+,CD3+CD4+,CD3+CD8+,CD3+CD4+/CD3+CD8+ (ratio only) lymphocyte percentages and absolute counts. - AQUIOS Tetra-2+ Panel Monoclonal Antibody Reagent Total CD3+, CD3-CD19+,CD3-CD56+ and/or CD16+ lymphocyte percentages and absolute counts. AQUIOS Flow Cytometry Software may be run on an independent computer workstation for off-line analysis of results generated by the AQUIOS CL Flow Cytometer with the monoclonal antibody reagents listed above. The off-
Similarities
	PredicateFACSCalibur		Device
Characteristic	CaliBRITE APC beads CaliBRITE 4 kit (unlabeled, FITC-, PE-, PerCP- and APC- labeled CaliBRITE beads), and FACSComp software	CaliBRITE 3 Color and FACSComp	AQUIOS CL Flow Cytometer
			line analysis must be performed in accordance with the product labeling.
Device Classification	Class II864.5220		Same
Sample Analysis	Principle of analysis – Flow cytometricDetection hardware – Lasers, fluidics, optics, electronicsSample analysis pathway		Same
Optics	Laser light delivered by mirrors, prisms, and lensesEmitted light delivered by mirrors		Same
	Software functionality to allow –
Workstation	Patient data management – storage, review, reporting to LISControl data management – storage, review, reportingSystem configuration managementSystem service test and adjustment procedures		Same
	Differences
Characteristic	PredicateFACSCalibur	CaliBRITE 3 Color andFACSComp	Device
Characteristic	CaliBRITE APC beadsCaliBRITE 4 kit (unlabeled, FITC-, PE-, PerCP- and APC-labeled CaliBRITE beads), andFACSComp software		AQUIOS CL Flow Cytometer
Product Code	GKZ		OYE
SystemConfiguration	BD FACSCalibur with FACS Loader Option and BD FACStation™ Mac Procomputer and Cell Quest SW		AQUIOS System with Reagent Cart andAll-in-One Touch Screen Computerloaded with AQUIOS System Software,Database, and Tetra Tests (Tetra 1, Tetra2, and Tetra Combo)
OperatingSystem	MAC OS 9		Microsoft Windows 7
Controllingsoftware	FACSComp		AQUIOS System Software
Lasers	Blue - 488 nm argon ionRed - 635 nm diode laser		Blue - 488 nm Blue Laser (20mW)
SamplePreparation	Manual		Automated On-board
Lyse Timing	15 minute incubation in the dark at room temperature.		Less than or equal to 3 minutes in thedark at room temperature
Lyse Reagents	BD FACS lysing solution		AQUIOS Lysing Reagent Kit
MonoclonalAntibodyAssay	BD Trucount tubesTwo tube antibody reagent assay:CD3-FITC/CD8-PE/CD45-PerCP/CD4-APC		Two tube monoclonal antibody reagentassay:o CD45-FITC/CD4-RD1/CD8-
Differences
	PredicateFACSCalibur		Device
Characteristic	CaliBRITE APC beadsCaliBRITE 4 kit (unlabeled,FITC-, PE-, PerCP- and APC-labeled CaliBRITE beads), andFACSComp software	CaliBRITE 3 Color andFACSComp	AQUIOS CL Flow Cytometer
	◦ CD3-FITC/CD16-PE+CD56-PE/CD45-PerCP/CD19-APC		ECD/CD3-PC5◦ CD45-FITC/CD56+CD16-RD1/CD19-ECD/CD3-PC5
SpecimenIntroduction	N/A		Specimens are loaded in the Autoloaderusing cassettes / or the single loader assingle specimen tubes
PreparedSampleIntroduction	FACS Loader (K953302)		96 well plate
SpecimenIdentification	Manual entry into Worklist		Barcode - positive sample identificationor manual entry
PreparedSampleIdentification	Worklist and Carousel position		Software tracked by well position.
Standardization	Automatic voltage/gain adjustments Compensation matrix calculation		A standardization check is performed oneach control run
SampleAnalysis	Manual gating of cellular populations per instrument IFU or automated gatingof cellular populations with BD Multiset software		Automated gating of cellular populationby AQUIOS System Software. Thesoftware only allows regionreadjustments by the user
Count Method	Cellular events are compared to bead events.	Automated method: Absolute counts are determined by MultiSETTM software.Manual data analysis using software such as CellQuestTM, absolute counts are	Syringe based volumetric method usingAQUIOS System Software
Differences
	PredicateFACSCalibur	Device
Characteristic	CaliBRITE APC beadsCaliBRITE 4 kit (unlabeled, FITC-, PE-, PerCP- and APC-labeled CaliBRITE beads), andFACSComp software	AQUIOS CL Flow Cytometer
	determined by division of the number of positive cellular events by thenumber of bead events multiplied by the TruCOUNT bead concentration.
PhotomultiplerTubes (PMTs) /Colors	FSC DiodeSSC 488/104 FL Detectors:• FL1 530/30 nm• FL2 585/42 nm• FL3 670 (LP) nm• FL4 661/16 nm	FS Solid State DetectorSS Solid State Detector – (488LP Dichroic)4 FL Detectors:• 525nm BP Filter• 575nm BP Filter• 620nm BP Filter• 675nm BP Filter
	Detectors/Parameters	Six (FS, SS, FL1-FL4)	Seven (EV, FS, SS, FL1 – FL4)
	Off-lineanalysissoftware	BD Multiset and BD Cell Quest Pro	Stand-alone workstations with AQUIOSSystem Software to review data offline
	Similarities
	Characteristic	Predicate		Device
		MultiTESTCD3/CD8/CD45/CD4	MultitestCD3/CD16+56/CD45/CD19Reagent and Multitest IMK KitLysing Solution with BDTruCount Tubes (PREDICATE)	AQUIOS Tetra-1 and AQUIOS Tetra-2+
Intended Use*	The BD Multitest IMK kit is a four-color directimmunofluorescence reagent kit for use with a suitably equippedflow cytometer to identify and determine the percentages andabsolute counts of the following mature human lymphocytesubsets in erythrocyte-lysed whole blood: T lymphocytes(CD3+), B lymphocytes (CD19+), helper/inducer T lymphocytes(CD3+CD4+), suppressor/cytotoxic T lymphocytes(CD3+CD8+), and natural killer (NK) lymphocytes (CD3-CD16+and/or CD56+). BD Trucount tubes are used for determiningabsolute counts. BD Multitest reagents and BD Trucount tubescan be used with the BD FACS Loader.* Only similar parameters of both devices are presented here.Refer to DxH 800 SE table for other parameters.		AQUIOS Tetra-1 Panel and AQUIOS Tetra-2+ Panelmonoclonal antibody reagents are for use on theAQUIOS CL Flow Cytometer with peripheral wholeblood for immunophenotyping. These reagents areindicated for use in the immunologic assessment ofpatients having, or suspected of having, immunedeficiency. These reagents provide identification andenumeration of;- AQUIOS Tetra-1 Panel Monoclonal AntibodyReagent• Total CD3+, CD3+CD4+,CD3+CD8+,CD3+CD4+/CD3+CD8+ (ratio only)lymphocyte percentages and absolutecounts.- AQUIOS Tetra-2+ Panel MonoclonalAntibody Reagent• Total CD3+, CD3-CD19+, CD3-CD56+
Similarities
	Predicate		Device
Characteristic	MultiTESTCD3/CD8/CD45/CD4	MultitestCD3/CD16+56/CD45/CD19Reagent and Multitest IMK KitLysing Solution with BDTruCount Tubes (PREDICATE)	AQUIOS Tetra-1 and AQUIOS Tetra-2+
DeviceClassification	Class II		Same
Reagent Assay	BD MultitestCD3/CD8/CD45/CD4reagent provided in 1 ml ofbuffered saline with 0.1%sodium azide	BD MultitestCD3/CD16+CD56/CD45/CD19reagent provided in 1 ml ofbuffered saline with 0.1% sodiumazide	Two test monoclonal antibody reagent assay:o CD45/CD4/CD8/CD3o CD45/CD56+CD16/CD19/CD3

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510(k) Submission for AQUIOS CL Flow Cytometry System Section 5: 510(k) Summary

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AQUIOS Tetra-1 and Tetra-2+ Device Comparison Tables:

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Differences
	Predicate		Device
Characteristic	MultiTESTCD3/CD8/CD45/CD4	MultitestCD3/CD16+56/CD45/CD19Reagent and Multitest IMK KitLysing Solution with BDTruCount Tubes (PREDICATE)	AQUIOS Tetra 1 and AQUIOS Tetra-2+
Product Code	GKZ		OYE
InstrumentQuality ControlTechniques	BD Multi-Check control, BD Multi-Check CD4 Low control andCaliBRITE Beads		AQUIOS Immuno-Trol and AQUIOS Immuno-TrolLow
AntibodyIncubationTime	15 minutes in the dark at room temperature		15-45 minutes in the dark at room temperature

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Similarities
Characteristic	PredicateUniCel DxH 800	DeviceAQUIOS Tetra 1 and AQUIOS Tetra-2+
Intended Use*	The UniCel® DxH 800 Analyzer is a quantitative multi-parameter, automated hematology analyzer for in vitro diagnostic use in screening patient populations found in clinical laboratories.The UniCel® DxH 800 Analyzer identifies and enumerates the parameters indicated below on the following sample types:Whole Blood (Venous) WBC, LY%, LY# * Only similar parameters of both devices are presented here	AQUIOS Tetra-1 Panel and AQUIOS Tetra-2+ Panel monoclonal antibody reagents are for use on the AQUIOS CL Flow Cytometer with peripheral whole blood. These reagents are indicated for use in the immunologic assessment of patients having, or suspected of having, immune deficiency. These reagents provide identification and enumeration of ;AQUIOS Tetra-1 Panel Monoclonal Antibody Reagent CD45+ absolute count CD45+ Low SS (lymphocytes) percentage and absolute count AQUIOS Tetra-2+ Panel Monoclonal Antibody Reagent CD45+ absolute count CD45+ Low SS (lymphocytes) percentage and absolute count
DeviceClassification	Class II	Same
Specimen AgeClaim	24 hours room temperature	Same
Similarities
Characteristic	PredicateUniCel DxH 800	DeviceAQUIOS Tetra 1 and AQUIOS Tetra-2+
SamplePreparation	Whole blood sample diluted with reagents	Same
Quality ControlTechniques	Daily Instrument ChecksCommercial ControlsInter-laboratory Quality Assurance Program (IQAP)	Daily Instrument ChecksCommercial ControlsInter-laboratory Quality Assurance Program (IQAP)
SamplingMechanism	Single aspiration probe used for all sampling.Single-tube presentation – open and closed vial sampling – specimen manually mixedAutomated presentation – closed vial sampling from five-position cassette accepting a variety of defined specimen tubes. Cassette containing specimens mixed prior to starting sampling and between specimens.	Same
Mechanismsfor Processing	Mechanisms to achieve process of :- automated cassette transportation and specimen mixing (by rocking)- sample aspiration- sample preparation- sample and reagent presentation to analytical modules- sample analysis- raw data collection- algorithmic processing- data reporting	Same
Differences
Characteristic	Predicate	Device
Intended Use	The UniCel® DxH 800 Analyzer is a quantitative multi-parameter, automated hematology analyzer for in vitrodiagnostic use in screening patient populations found inclinical laboratories.The UniCel® DxH 800 Analyzer identifies and enumeratesthe parameters indicated below on the following sampletypes:Whole Blood (Venous and Capillary) RBC, HGB, HCT, MCV, MCH, MCHC, RDW,RDW-SD, PLT, MPV, NE%, NE#, MO%,MO#, EO%, EO#, BA%, BA#, NRBC%,NRBC#, RET%, RET#, MRV, IRF Pre-Diluted Whole Blood (Venous and Capillary) WBC, RBC, HGB, HCT, MCV, MCH, MCHC,RDW, RDW-SD, PLT, MPV Body Fluids (cerebrospinal, serous and synovial) TNC and RBC	AQUIOS Tetra-1 and AQUIOS Tetra-2+AQUIOS Tetra-1 Panel and AQUIOS Tetra-2+ Panelmonoclonal antibody reagents are for use on theAQUIOS CL Flow Cytometer with peripheral wholeblood. These reagents are indicated for use in theimmunologic assessment of patients having, or suspectedof having, immune deficiency. These reagents provideidentification and enumeration of;AQUIOS Tetra-1 Panel Monoclonal AntibodyReagent Total CD3+, CD3+CD4+,CD3+CD8+,CD3+CD4+/CD3+CD8+ (ratio only)lymphocyte percentages and absolute counts. AQUIOS Tetra-2+ Panel Monoclonal AntibodyReagent percentage and absolute count Total CD3+, CD3-CD19+, CD3-CD56+and/or CD16+ lymphocyte percentages andabsolute counts. AQUIOS Flow Cytometry Software may be run on anindependent computer workstation for off-line analysisof results generated by the AQUIOS CL Flow Cytometerwith the monoclonal antibody reagents listed above. Theoff-line analysis must be performed in accordance withthe product labeling.
Differences
	Predicate	Device
Characteristic	UniCel DxH 800	AQUIOS Tetra-1 and AQUIOS Tetra-2+
Product Code	GKZ	OYE
Specimen AgeClaim	48 hours refrigerated	Should not be refrigerated.
WBC, RBC,MCV, Plt, TNC	Aperture impedance (Coulter® Principle)	Aperture impedance (Coulter® Principle) and flowcytometry analysis
PerformanceCharacteristics	Quantitative test for RBC, HGB, HCT, MCV, MCHC,RDW, RDW-SD, PLT, MPV, NE%, NE#, MO%, MO#,EO%, EO#, BA%, BA#, NRBC%, NRBC#, RET%, RET#,MRV, IRF, TNC	Total CD3+, CD3+CD4+,CD3+CD8+,CD3+CD4+/CD3+CD8+ (ratio only),CD3-CD19+, CD3-CD56+ and/or CD16+ lymphocytepercentages and absolute counts.
Reagents todetermineLymphocytes	Coulter® DxH DiluentCoulter® DxH Diff Pack	AQUIOS Tetra-1 - CD45-FITC, B3821F4A; CD4-RD1,SFCI12T4D11; CD8-ECD, SFCI21Thy2D3; CD3-PC5,UCHT1AQUIOS Tetra-2+ - CD45-FITC, B3821F4A; CD3-PC5,UCHT1; CD56+CD16-RD1, N901/NKH-1;CD19-ECD,J3-119AQUIOS Lysing Reagent KitAQUIOS Sheath
Reagents todetermine WBC	Coulter® DxH Diff PackCoulter® DxH Cell Lyse	AQUIOS Tetra-1 - CD45-FITC, B3821F4A; CD4-RD1,SFCI12T4D11; CD8-ECD, SFCI21Thy2D3; CD3-PC5,UCHT1AQUIOS Tetra-2+ - CD45-FITC, B3821F4A; CD3-PC5,UCHT1; CD56+CD16-RD1, N901/NKH-1;CD19-ECD,13-119AQUIOS Lysing Reagent KitAQUIOS Sheath
Controls	Coulter® 6C Cell Control	AQUIOS Immuno-Trol Cells
Calibrators	Coulter® Latron™ CP-X ControlCoulter® S-CAL® Calibrator Kit	AQUIOS Immuno-Trol Low CellsFactory/Service calibrated
Similarities
	Predicate	Device
Characteristic	Immuno-Trol Control Cells(Predicate)	Immuno-Trol Low Cells(Predicate)	AQUIOS Immuno-Trol and Immuno-Trol Low
Intended Use	IMMUNO-TROL™ Control Cells(Immuno-Trol) is an assayed,lysable whole blood qualitycontrol product forimmunophenotyping analysisusing monoclonal antibodyreagents and flow cytometry. Itprovides a positive cell controlthat is processed in the samemanner as a whole blood sample.This allows verification of reagentperformance and the methodsused for staining targeted cells,lysing erythrocytes, and analyzingsamples by flow cytometry.	IMMUNO-TROL™ Low Cells is anassayed, lysable whole blood qualitycontrol product forimmunophenotyping analysis usingmonoclonal antibody reagents andflow cytometry. It provides a positivecell control that is processed in thesame manner as a whole bloodsample.	AQUIOS Immuno-TrolAQUIOS IMMUNO-TROL™ Cells areassayed, lysable whole blood qualitycontrol product for immunophenotypinganalysis using monoclonal antibodyreagents and flow cytometry. It provides apositive cell control that is processed in thesame manner as a whole blood sample.This allows verification of instrument andreagent performance. It also verifies themethods used for staining targeted cells,lysing erythrocytes, and analyzing samplesby the AQUIOS CL Flow Cytometer.AQUIOS Immuno-Trol LowAQUIOS IMMUNO-TROL™ Low Cellsare assayed, lysable whole blood qualitycontrol product for immunophenotypinganalysis using monoclonal antibodyreagents and flow cytometry. It provides a

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510(k) Submission for AQUIOS CL Flow Cytometry System Section 5: 510(k) Summary

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Similarities
	Predicate		Device
Characteristic	Immuno-Trol Control Cells(Predicate)	Immuno-Trol Low Cells(Predicate)	AQUIOS Immuno-Trol and Immuno-Trol Low
DeviceClassificationand ProductCode	Class II, JPK		Same

Differences
Characteristic	Predicate	Device
	Immuno-Trol Control Cells(Predicate)	AQUIOS Immuno-Trol and Immuno-Trol Low
	Immuno-Trol Low Cells(Predicate)
AssayParameters	CD3+ count, %	Same except with the addition of;
	CD3+/CD4+ count, %	CD3-/(CD56+16)+ count, %
	CD3+/CD8+ count, %	CD45+ count
	CD3-/CD56+ count, %	CD45+ Low SS count, %
	CD19+ count, %	CD3 Separation Quotient
		CD4 Separation Quotient
		CD8 Separation Quotient
		CD56+16 Separation Quotient
		CD19 Separation Quotient
		CD45 Lymph/High SS Cells SeparationQuotient
		SS Lymph/High SS Cells SeparationQuotient
		FS Lymph/High SS Separation Quotient

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Study	Testing Approach	FDA GuidanceDocuments	Standards/ References	Testing Results
FluorescenceLinearity	Verify fluorescencedetection is linear usingstandard AQUIOS Tetrasettings.	None	None	Linearity of fluorescence measurementswas demonstrated.
ElectronicVolumeLinearity	Verify linearity ofElectronic Volumemeasurements at thestandard AQUIOS Tetrasettings.	None	None	Linearity of Electronic Volumemeasurements was demonstrated.
LaserPerformanceCharacteristics	Verify stability of thelaser performance of theAQUIOS CL flowcytometer over time.	None	None	Analysis of the data collecteddemonstrates that the AQUIOS CL laserperformance is stable over time.
AnalyzerCarryover	To verify carryover forwhole blood meetsperformancespecifications.	Special ControlsGuidance Document:Premarket Notificationsfor AutomatedDifferential CellCounters for Immatureor Abnormal BloodCells - Carryover(Section 12)	CLSI H26-A2; FDA StandardsRecognition #7-210	Analysis of the data collecteddemonstrates that the AQUIOS CLmeets the whole blood carryoverperformance requirements.
Study	Testing Approach	FDA GuidanceDocuments	Standards/ References	Testing Results
InstrumentSettings Stability	Testing to demonstratestability of the AQUIOSinstrument settings tosupport adequacy of thequality controlmethodology as anindicator of instrumentperformance.	None	None	Analysis of the data collecteddemonstrate that the stability of theAQUIOS CL instrument settingssupport the quality controlmethodology.
Gravimetrics	Evaluate accuracy andprecision of dispensingAQUIOS lysing andantibody reagents, andspecimenaspiration/dispense.	None	None	Analysis of the data collecteddemonstrates accuracy and precision ofdispensing AQUIOS lysing andantibody reagents, and specimenaspiration/dispense.
Comparability -Collection TubePerformance	Evaluate tubecharacteristics when usedon the AQUIOS system.Study will includeverification of thedispensed specimenvolumes and minimumrequired aspirationvolumes across thedifferent tube types.Results obtained with thedifferent tube types willbe compared at theminimum and maximumnumber of pierces.	None	CLSI EP15-A2, UserVerification of Performancefor Precision and Trueness;Approved Guideline – SecondEdition. (FDA RecognitionNumber 7-153)	Analysis of the data collected verifiedthe specimen tube recommendations inproduct labeling.
Study	Testing Approach	FDA GuidanceDocuments	Standards/ References	Testing Results
Assay Linearity	Verify the linear range ofthe absolute values foreach lymphocyte subsetpopulations on theAQUIOS CL FlowCytometry System.	Special ControlsGuidance Document:Premarket Notificationsfor AutomatedDifferential CellCounters for Immatureor Abnormal BloodCells - Linearity(Section 11)	Evaluation of the Linearity ofQuantitative MeasurementProcedures: A StatisticalApproach; ApprovedGuideline; April 2003. CLSIEP06-A; FDA StandardsRecognition #7-193	Analysis of the data collecteddemonstrates that the AQUIOS CLmeets the linearity performancerequirements.
Assay Carryover	To verify carryover ofwhole blood and reagentson the AQUIOS CL meetsperformancespecifications.	Special ControlsGuidance Document:Premarket Notificationsfor AutomatedDifferential CellCounters for Immatureor Abnormal BloodCells - Carryover(Section 12)	Validation, Verification, andQuality Assurance ofAutomated HematologyAnalyzers, Approved Standard- 2nd Edition; June 2010;CLSI H26-A2; FDA StandardsRecognition #7-210	Analysis of the data collecteddemonstrates that the AQUIOS CLmeets carryover performancerequirements.
DetectionCapability	To verify that theAQUIOS CL meets theperformance requirementsfor Limit of Blank (LoB),Lower Limit of Detection(LLoD), Lower Limit ofQuantitation (LLoQ).	None	Evaluation of DetectionCapability for ClinicalLaboratory MeasurementProcedures; ApprovedGuideline - Second Edition;CLSI EP17-A2; FDAStandards Recognition #7-233	Analysis of the data collecteddemonstrates that the AQUIOS CLmeets the performance requirements forLoB, LLoD, and LLoQ in whole blood.
Study	Testing Approach	FDA GuidanceDocuments	Standards/ References	Testing Results
Specimen andPrepared SampleStability	Stability of whole bloodsamples will be evaluatedto verify specimen andprepared sample stabilityclaims.	None	None	Analysis of the data collecteddemonstrates that the AQUIOS CLFlow Cytometry System meets therequirements for specimen and preparedsample stability.
MethodComparison	To evaluate bias betweenthe subject device versusthe predicate.	Special ControlsGuidance Document:Premarket Notificationsfor AutomatedDifferential CellCounters for Immatureor Abnormal BloodCells - Accuracy(Section 8)	Validation, Verification, andQuality Assurance ofAutomated HematologyAnalyzers, Approved Standard– 2nd Edition; June 2010;CLSI H26-A2; FDA StandardsRecognition # 7-210CLSI EP09-A3, MethodComparison and BiasEstimation Using PatientSamples; Approved Guideline--Second Edition (InterimRevision)	Analysis of the data collecteddemonstrates that the AQUIOS CLFlow Cytometry System meets theperformance requirements whencompared to the predicate device.
Precision - LongTermImprecision	Demonstrate systemimprecision using controlmaterial as a surrogate fora stabilized sample	Special ControlsGuidance Document:Premarket Notificationsfor AutomatedDifferential CellCounters for Immatureor Abnormal BloodCells - Precision(Section 9)	CLSI EP5-A2, Evaluation ofPrecision Performance ofQuantitative MeasurementMethods; Approved Guideline– Second Edition.	Analysis of the data collecteddemonstrates that the AQUIOS CLmeets performance requirements forLong Term Imprecision.
Study	Testing Approach	FDA GuidanceDocuments	Standards/ References	Testing Results
Precision –Whole BloodRepeatability	Evaluate sampleimprecision using aprecision profile approachto estimate repeatability atvarious medical decisionlevels and datapercentiles.	Special ControlsGuidance Document:Premarket Notificationsfor AutomatedDifferential CellCounters for Immatureor Abnormal BloodCells - Precision(Section 9)	CLSI H26-A2, Validation,Verification, and QualityAssurance of AutomatedHematology Analyzers	Analysis of the data collecteddemonstrates that the AQUIOS CLmeets performance requirements forWhole Blood Repeatability.
Comparability(Equivalency) –Anticoagulant,SamplePresentationMode, Open andClosed VialSampling Modes,Test PanelEquivalency	Provide data onequivalency ofanticoagulants, modes,and panels for use onAQUIOS system.	Special ControlsGuidance Document:Premarket Notificationsfor AutomatedDifferential CellCounters for Immatureor Abnormal BloodCells - Accuracy(Section 8)	CLSI EP09-A3, MethodComparison and BiasEstimation Using PatientSamples; Approved Guideline--Second Edition (InterimRevision)	For all conditions evaluated, the datacollected demonstrates that theAQUIOS CL within-methodcomparisons are equivalent.
Adult ReferenceIntervals	Establish adult referenceintervals for lymphocytesubset analysis on theAQUIOS system.	Special ControlsGuidance Document:Premarket Notificationsfor AutomatedDifferential CellCounters for Immatureor Abnormal BloodCells – ReferenceValues (Section 14)	C28-A3c, Defining,Establishing, and VerifyingReference Intervals in theClinical Laboratory	Adult reference intervals wereestablished and confirmed to beconsistent with published values for T,B, and NK lymphocyte subsets.
Study	Testing Approach	FDA GuidanceDocuments	Standards/ References	Testing Results
Stability	Stability testing willdemonstrate the shelf lifeof the reagents.	None	CLSI EP25-A - Evaluation ofStability of In Vitro DiagnosticReagents. Approved Guideline	Analysis of the data collecteddemonstrates that the AQUIOS Tetra-1and Tetra-2+ reagents meet performancerequirements in support of the product'sstability claims.
Lot Variability	Variability of multiplelots of material wasincorporated into theoverall system variabilityassessed in the clinicaltesting.	None	CLSI EP5-A2, Evaluation ofPrecision Performance ofQuantitative MeasurementMethods; Approved Guideline- Second Edition.	Analysis of the data collecteddemonstrates that the AQUIOS Tetra-1and Tetra-2+ have acceptable lotvariability performance.

Summary of AQUIOS CL Flow Cytometer Performance Testing

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Summary of AQUIOS Tetra-1 and Tetra-2+ Performance Testing:

Summary of AQUIOS Immuno-Trol and Immuno-Trol Low Performance Testing:

Study	Testing Approach	FDA GuidanceDocuments	Standards/ References	Testing Results
Assay ValueAssignment andExpected Ranges	Determine the expectedranges (count, percent,separation quotient) forboth levels of control aswell as the process toassign these values toeach lot of material.	None	None	Assay value assignment process andexpected ranges were established andverified based on analysis of the datacollected.
Stability	Stability testing willdemonstrate the shelf lifeof the reagents.	None	CLSI EP25-A – Evaluation ofStability of In Vitro DiagnosticReagents. Approved Guideline	Analysis of the data collecteddemonstrates that the AQUIOSImmuno-Trol and Immuno-Trol Lowmeet performance requirements in

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Study	Testing Approach	FDA GuidanceDocuments	Standards/ References	Testing Results
				support of the product's stability claims.
Lot Variability	Verify lot variability ofAQUIOS Immuno-Troland Immuno-Trol Low.	None	CLSI EP5-A2, Evaluation ofPrecision Performance ofQuantitative MeasurementMethods; Approved Guideline- Second Edition.	Analysis of the data collecteddemonstrates that the AQUIOSImmuno-Trol and Immuno-Trol Lowhave acceptable lot variabilityperformance.

Summary of AQUIOS Lysing Reagent Kit Performance Testing:

Study	Testing Approach	FDA GuidanceDocuments	Standards/ References	Testing Results
Stability	Stability testing willdemonstrate the shelf lifeof the reagents.	None	CLSI EP25-A - Evaluation ofStability of In Vitro DiagnosticReagents. Approved Guideline	Analysis of the data collecteddemonstrates that the AQUIOS Lysereagents meet performance requirementsin support of the product's stabilityclaims.
Lot Variability	Variability of multiplelots of material wasincorporated into theoverall system variabilityassessed in the clinicaltesting.	None	CLSI EP5-A2, Evaluation ofPrecision Performance ofQuantitative MeasurementMethods; Approved Guideline- Second Edition.	Analysis of the data collecteddemonstrates that the AQUIOS Tetra-1and Tetra-2+ have acceptable lotvariability performance.

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Substantial Equivalence Conclusion to Demonstrate Safety, Effectiveness & Equivalent Performance to Predicate:

The AQUIOS CL Flow Cytometry System, that is the subject of this submission, in concert with the conclusions drawn from the performance testing discussed above demonstrate that when compared to the predicate device is as safe, as effective, and meets the performance acceptance criteria.

In summary, the AQUIOS CL Flow Cytometry System as described in this submission is substantially equivalent in terms of safety and effectiveness to its predicate devices.

. The AQUIOS CL Flow Cytometer and AQUIOS Tetra-1 and Tetra-2+ reagents are substantially equivalent to the FACSCalibur CaliBRITE 4 kit and FACSComp Software as well as the UniCel DxH 800, where applicable.
The AQUIOS Immuno-Trol and Immuno-Trol Low are substantially equivalent to Immuno-Trol Cells and Immuno-Trol Low Cells.

This summary of safety and effectiveness is being submitted in accordance with the requirements of the Safe Medical Device Act of 1990 and the implementing regulation 21 CFR 807.92.

Regulation Number and Section

§ 864.5220 Automated differential cell counter.

(a)
Identification. An automated differential cell counter is a device used to identify one or more of the formed elements of the blood. The device may also have the capability to flag, count, or classify immature or abnormal hematopoietic cells of the blood, bone marrow, or other body fluids. These devices may combine an electronic particle counting method, optical method, or a flow cytometric method utilizing monoclonal CD (cluster designation) markers. The device includes accessory CD markers.(b)
Classification. Class II (special controls). The special control for this device is the FDA document entitled “Class II Special Controls Guidance Document: Premarket Notifications for Automated Differential Cell Counters for Immature or Abnormal Blood Cells; Final Guidance for Industry and FDA.”