81 FR 46054 - Flow Cytometry Quantitation Consortium

DEPARTMENT OF COMMERCE
National Institute of Standards and Technology

Federal Register Volume 81, Issue 136 (July 15, 2016)

The National Institute of Standards and Technology (NIST), an agency of the United States Department of Commerce, is establishing the Flow Cytometry Quantitation Consortium and invites organizations to participate in this Consortium. The Consortium will develop reference materials including reference fluorophore solutions and biological reference materials, reference data and reference methods for assigning equivalent number of reference fluorophores (ERF) values and for assessing the associated uncertainties and utilities. Participation in this Consortium is open to all eligible organizations, as described below.

[Federal Register Volume 81, Number 136 (Friday, July 15, 2016)]
[Notices]
[Pages 46054-46055]
From the Federal Register Online  [www.thefederalregister.org]
[FR Doc No: 2016-16761]


-----------------------------------------------------------------------

DEPARTMENT OF COMMERCE

National Institute of Standards and Technology


Flow Cytometry Quantitation Consortium

AGENCY: National Institute of Standards and Technology, Department of 
Commerce.

ACTION: Notice; request for information.

-----------------------------------------------------------------------

SUMMARY: The National Institute of Standards and Technology (NIST), an 
agency of the United States Department of Commerce, is establishing the 
Flow Cytometry Quantitation Consortium and invites organizations to 
participate in this Consortium. The Consortium will develop reference 
materials including reference fluorophore solutions and biological 
reference materials, reference data and reference methods for assigning 
equivalent number of reference fluorophores (ERF) values and for 
assessing the associated uncertainties and utilities. Participation in 
this Consortium is open to all eligible organizations, as described 
below.

DATES: NIST will accept responses for participation in this Consortium 
on an ongoing basis. The Consortium's activities will commence on 
August 15, 2016 (``Commencement Date''). Acceptance of participants 
into the Consortium after the Commencement Date will depend on the 
availability of NIST resources.

ADDRESSES: Information in response to this notice and requests for 
additional information about the Consortium can be directed via mail to 
the Consortium Manager, Dr. Lili Wang, Biosystems and Biomaterials 
Division of NIST's Material Measurement Laboratory, 100 Bureau Drive, 
Gaithersburg, Maryland 20899-8312, or via electronic mail to 
[email protected].

FOR FURTHER INFORMATION CONTACT: For further information about 
partnership opportunities or about the terms and conditions of NIST's 
Cooperative Research and Development Agreement (CRADA), please contact 
Honeyeh Zube, CRADA and License Officer, National Institute of 
Standards and Technology's Technology Partnerships Office, by mail to 
100 Bureau Drive, Mail Stop 2200, Gaithersburg, Maryland 20899, by 
electronic mail to [email protected], or by telephone at (301) 975-
2209.

SUPPLEMENTARY INFORMATION: Flow cytometry is a widely used technique 
for a single cell and particle analysis. It is an essential tool for 
immunological research, drug and device development, clinical trials, 
disease diagnosis, and therapy monitoring. The annual expenditure on 
flow cytometry-related diagnostics is upwards of $1.2 Billion and 
growing at more than 10 percent per year, testifying to the economic 
importance of this technology. The measurements made on the different 
instrument platforms at different times and locations, however, cannot 
be compared accurately, which makes diagnostic decisions unreliable and 
slows down advances in biomedical research. In response to this 
limitation, NIST and International Society for Advancement of Cytometry 
(ISAC) have developed a methodology to implement quantitation in flow 
cytometry. The first step is to calibrate the fluorescence signal from 
microparticles in terms of a unit of equivalent number of reference 
fluorophores (ERF) on three laser excitations, 405 nm, 488 nm, and 633 
nm. The ERF unit gives the number of reference fluorophores in solution 
which produce the same fluorescence signal as a single dyed 
microsphere.
    The second step uses a biological cell, with known number of 
specific biomarkers, as a reference material to translate the ERF unit 
to a unit of antibodies bound per cell (ABC). The ABC unit is most 
relevant to immunological measurements. To support the calibration of 
microparticles in terms of ERF, NIST has developed standard reference 
material (SRM 1934), which includes four solutions of fluorophore: 
Fluorescein, Nile Red, Coumarin 30 and Allophycocyanin. Microparticles 
that have been assigned ERF values using SRM 1934 will enable the 
calibration and characterization of flow cytometers, and the 
standardization of the fluorescence intensity scale in quantitative ERF 
units. The results of the collaboration under this Consortium will 
allow the industry to further research, develop and adopt reference 
fluorophore solutions for other laser excitations and reference 
material standards recommended by the expert user community.
    NIST is establishing this five-year Consortium to collaborate with 
manufacturers of microparticles to develop methodologies for assigning 
ERF values for the microparticles provided to NIST under the scope of 
the Consortium. The results from this Consortium will also allow NIST 
to develop the capability that NIST would require to provide a 
calibration service.
    The certificate of analysis for NIST SRM 1934 and NIST's finalized 
standard operating procedure (SOP) for assigning ERF value will be used 
for performing the ERF value assignments for participants' 
microparticles. This SOP includes four steps and is published at J. 
Res. Natl. Inst. Stand. Technol. 121: 269-286 (2016). As described in 
the SOP, the ERF value of the major microparticle population is 
calculated on the basis of the ratio of mean fluorescence intensity 
values of the major microparticle population to all microparticle 
populations.
    A summary of the ERF value assignments will include ERF values of 
major microparticle populations, associated combined uncertainties per 
laser excitation, and reference fluorophore. The combined uncertainty 
will be derived from all steps of the ERF value assignment, from 
weighing reference solutions, spectrofluorimeter calibration, CCD 
response calibration, microparticle concentration measurements by flow 
cytometer and light obscuration, and measurement of the emission 
spectrum of microparticles to determine ERF values for major 
microparticle populations. NIST will also share with each participant 
any digital emission spectral data of the major microparticle 
populations. In addition, a participant may request reports for 
specific ERF value assignments for its microparticles under this 
Consortium. NIST intends to

[[Page 46055]]

publish anonymized results of the research under this Consortium. In 
accordance with 15 U.S.C. 3710a(c)(7)(B), NIST will withhold from 
public disclosure the data that specifically identifies a participant's 
microparticles for a period of five (5) years from the date any ERF 
values are generated, or until the participants grants NIST permission 
to disclose such data. NIST will not require the participants to pay a 
membership fee to participate in this Consortium. NIST will, however, 
require participants to contribute funds to reimburse NIST for the 
generation of any report requested by a participant for the ERF value 
assignments of participant's microparticles.
    Participation Process: Eligibility will be determined by NIST using 
the information provided by an organization in response to this notice 
based on the information requested below.
    An organization responding to this notice should provide the 
following information to NIST's Consortium Manager:
    (1) Type of microparticles: Optimal sizes of microparticles are 
from 2 to 10 microns. If there are needs of characterization and ERF 
value assignment to other size particles (<2 microns or >10 microns), 
the present standard operating procedure can be modified to accommodate 
the requests.
    (2) Type of Instrument: The Consortium is to assign ERF values for 
microparticles used primarily for flow cytometers. Any information 
about other instruments used by the organization is helpful to ensure 
that there is diversity in participants. For example, please indicate 
if the microparticles are used by the organization with fluorescence 
microscopes and spectrophotometers/spectrofluorimeters.
    (3) Experience in production and characterization of 
microparticles, antibodies, and biological cells, and analysis of large 
data sets.
    A responding organization should not include any business 
proprietary information in its response to this request for 
information. NIST will not treat any information provided in response 
to this request as proprietary information. NIST will notify each 
organization of its eligibility. In order to participate in this 
Consortium, each eligible organization must sign a Cooperative Research 
and Development Agreement (CRADA) for this Consortium. All participants 
to this Consortium will be bound by the same terms and conditions.

Kent Rochford,
Associate Director for Laboratory Programs.
[FR Doc. 2016-16761 Filed 7-14-16; 8:45 am]
 BILLING CODE 3510-13-P