JA_FACSAria Fusion_75N93020P01063_Redacted.pdf

blood specimens obtained from individuals enrolled in multiple clinical research protocols. A dedicated, reliable and enclosed cell sorter is urgently needed for this large collaborative effort to proceed. Other needs with the new instrument include investigating the immune parameters from cells sorted in other LIR priorities related to HIV research. To meet this need, LIR is requesting the purchase of a cell sorter with custom built biosafety cabinet to replace a 15-year old cell sorter that is no longer in reliable working condition. The LIR also has a need to limit the footprint of the instrument given limited space in the Biosafety Level 2/3 laboratories where the research is performed; the combination of the sorter with a custom-built biosafety cabinet achieves this need and provides the needed safety for working with human pathogens. In addition to the NIDDK collaboration, LIR is pursuing several lines of investigation on the immunity of HIV-infected individuals enrolled in numerous clinical research protocols. Among the specific needs is to sort HIV-specific B cells from participants in these protocols for the purpose of understanding their antibody response to HIV. LIR is also collaborating with other NIAID researchers where cell sorting is used to follow the course of immune deficiency diseases.

The LIR Immunopathogenesis Section currently owns one cell sorter, a BD Biosciences FACSAria IIu, purchased in 2004. The instrument is nearing the end of its life cycle and has been experiencing persistent failures especially with a feature called drop delay, one of the sorting parameters that must be adjusted prior to sorting and is a critical determinant of sort recoveries/purities. This failure jeopardizes research outcomes, either by yielding samples of low viability/specificity or by loss of specimen if failure cannot be fixed. Given that samples prepared for sorting cannot be stored for later processing, instrument failure invariably means loss of an entire experiment. LIR has spent years developing multiple protocols and testing parameters on the current, older, cell sorter, as well as identifying a wide breadth of reagents used in these protocols (i.e. staining antibodies to sort for molecules of interest). These protocols/parameters and reagents needs to be able to be directly implemented on the new cell sorter with limited confirmation testing.

Critically, the instrument required must provide seamless continuity between the existing cell sorter and the new instrument such that data produced on current instrumentation will be identical to that from the new instrument.

This is a commercial supplies requirement. Previous NIAID purchase for similar instruments were sole source, open market acquisitions:

Purchase Order No.: 75N93019P01251 Vendor: Becton, Dickinson and Company Period of Performance: 9/27/2019 – 4/17/2020 Amount: $2,109,663.75

Purchase Order No.: 75N93019P01245

Period of Performance: 9/25/2019 – 1/22/2020

Amount: $327,752.50

Purchase Order No.: HHSN272201801359P

Period of Performance: 9/18/2018 – 9/28/2018 Amount: $689,942.23

3. Description of the supplies or services required to meet the agency’s needs (including the estimated value).

a. Project title. BD FACSAria™ Fusion Cell Sorter and Biosafety Cabinet

b. Project description. This requirement includes an advanced cell sorter and biosafety solution with the following key specifications:

Cell sorter must offer a minimum of 4-way tube sorting capability and an automatic cell deposition unit for sorting into 96-well plates.

Cell sorter must have capability of achieving high sorting rates, demonstrating linear and accurate data acquisition at above 50,000 events per second.

Cell sorter must offer user-friendly optical alignment, laser alignment or laser focus to reduce instrument set-up time and operator interaction in the biosafety cabinet.

Must have a gel-coupled cuvette that collects four times as much excitation light compared to stream-in-air systems to maximize sensitivity in signal detection (Flow Cytometry Lens System).

Sensitivity should have minimum resolution <118MESF on FITC and 34MESF on PE at 90KhZ and 70psi using Sphero rainbow beads.

Cell sorter must not exceed dimensions of H 86.5 in, x W 50.2 in, x D 35.6 to fit in space available and biosafety cabinet

Cell sorter must be installed in an NSF-49 certified Biosafety cabinet with ULPA filtration system for live cell sorting.

Cell sorter must include an octagon shaped collection optics that increases the quality and quantity of the fluorescent signal. Fiber coupled from collection lens (Fluorescence Detection Instrument with Reflective Transfer Legs for Color Decimation).

Must have three lasers: 405nm 100mw, 488 nm 100mw and 637nm100mw.

Cell sorter must use digital technology from the preamp forward.

The digital electronics must use mathematical calculations in place of analog log amps to provide more linear quantitative measurements and high throughput.

Cell sorter must have the ability to achieve threshold on multiple parameters simultaneously. This is a power tool for detecting small particles or samples with large amounts of debris.

Must have the ability to establish signal-to-noise ratio from any laser.

Cell Sorter must include a System for Identifying Clusters in Scatter Plots using Smoothed Polygons with Optimal Boundaries.

Cell Sorter must have a cloning unit that can identify unique sort regions for different wells within the same 96 well plate with the ability to chill or heat the 96 well tray.

Cell sorter must have Index Sorting to identify sorted cells on scatter plot and cells in any well position on scatter plot.

Cell sorter must employ automatic sort setup technology that allows the operator to monitor side streams while sorting. This provides for a faster sort set-up providing the operator with a visual display of the sort set-up (Apparatus and Method for Verifying Drop Delay in A Flow Cytometer).

Cell sorter must have the ability to collect Height, Area and Width parameters simultaneously on all parameters.

Cell sorter must be able to utilize up to 6 solid state diode or DPSS lasers in various wavelengths and outputs lasers and up to 18 colors plus forward and side scatter.

Cell Sorter must utilize proprietary software that acquires and analyses in bi-exponential display format with FCS 2.0, 3.0 and 3.1 export of analyzed data.

Cell sorter must have proven instrument set-up and automated quality control software package that allows for automation in tracking instrumentation performance. It must monitor signal to noise ratio, minimal noise levels for all PMTs and parameters, and allow for feedback on the Q of the laser.

Cell Sorter must have automated compensation networks between all lasers, and all fluoresces parameters.

Cytometer must be serviceable by in-state factory trained engineers with 48 hour guaranteed on-site repair response time.

Cell Sorter must self-regulate during sorting and have the ability to shut down sample flow, sheath flow, and cover sorted materials in presence of gross shift in sorting parameters (Flow Sorter with Video-Regulated Droplet Spacing).

A redundant aerosol management system must be included in event of blower motor failure. The system must evacuate the sort collection chamber by trapping aerosolized particles with a 0.10 m size ultra-low penetrating air filter.

Cell sorter must offer seamless integration with existing FACSAria Cell sorter for ease of assay transfer between sorters and minimum training of operators with identical sample acquisition software

Requirement type.

Research & development (R&D) R & D support services Support services (non-R&D) Supplies/equipment Information technology (IT) Construction Architect-engineer (A & E) services

Design-build

Type of action.

New requirement Follow-on Other (specify): ______________________

Proposed contract/order type.

Firm-fixed-price Other fixed-price (specify, e.g., fixed-price award-fee, fixed-price incentive-fee): __________ Cost-plus-fixed-fee Other cost reimbursement (specify, e.g., cost-plus-award-fee, cost-plus-incentive-fee): ___________ Time and materials Indefinite delivery (specify whether indefinite quantity, definite quantity, or requirements): ___________ Other (specify): ___________ Completion Form Term form

Acquisition identification number. NOI-RML-2025189

c. Total estimated dollar value and performance/delivery period.

Total Dollar Value:

Period of Performance: August 24, 2020 – August 23, 2021

4. Identification of the statutory authority permitting other than full and open competition.

This acquisition is conducted under the authority of 41 United States Code (U.S.C.) 253(c)(__) as set forth in Federal Acquisition Regulation (FAR) 6.302- __.

This acquisition is conducted under the authority of 41 unite States Code (U.S.C) 3304(a)(1) as set forth in Federal Acquisition Regulation (FAR) 13.106-1(b)(2) and 13.501(a).

This acquisition is conducted under the authority of the Services Acquisition Reform Act of 2003 (41 U.S.C. 428a).

5. Demonstration that the proposed contractor(s) unique qualifications or the nature of the acquisition requires use of the authority cited.

a. Name and address of the proposed contractor(s).

BECTON DICKINSON AND COMPANY - BD Biosciences Business Unit One Becton Drive, Franklin Lakes, NJ 07417

6. Nature of the acquisition and proposed unique qualifications of the contractor(s). The LIR is proposing to procure a BD Biosciences FACSAria Fusion cell sorter with custom biosafety cabinet to replace their FACSAria IIu cell sorter manufactured by BD Biosciences. Utilizing the same equipment brand, reagents, and protocols will ensure consistent operations, functionality, and compatibility for successful support of critical research efforts. The FACSAria Fusion cell sorter is the only available cell sorter that provides all the sorting and analytical capabilities as well as technological advances described in the specifications, and thus BD Biosciences is the only vendor that can meet the salient characteristics of the requirement detailed in the specifications above.

Further, on the BD FACSAria Fusion cell sorter provides for uninterrupted continuity of research and immediate compatibility with existing software that is currently used at LIR, and only BD Biosciences can provide the maintenance support and services the LIR requires in order to maximize instrument operability.

LIR has used BD instrumentation for many years and currently relies on a BD FACSAria IIu for their cell sorting needs. Thus, a significant amount of protocols and testing parameters have been developed in the lab and published by researchers. Procurement of a new BD instrument will allow LIR staff and users to use these existing protocols and parameters whereas switching to a different manufacturer would require development of new protocols and parameters which would result in unacceptable delays. In addition, ongoing experiments have been optimized for particular characteristics of the BD platform. Using a different platform would require re-testing and re-developing protocols and testing parameters and identifying new reagents and fluorochrome combinations (and re-optimize the appropriate dilutions) in order to achieve a similar level of sorting capability as LIR’s older instrument – FACSAria IIu. This would waste a year’s worth of employee and instrument time and greatly hinder LIR’s research progress and output. The new cell sorter will be located in a Biosafety Level 2/3 laboratory and it is imperative that this instrument provides the needed safety for working with human pathogens such as HIV. There are multiple cell sorters in the marketplace that offer integrated cabinet options for aerosol management;

however, only BD has the added feature of an enclosed sort collection system where most of the aerosols are generated due to the highly pressurized nature of all sorters. This is an important feature for LIR researchers as they plan to sort samples from patient infected with highly transmissible or deadly pathogens such as HIV and the novel coronavirus.

7. Description of the efforts made to ensure that offers are solicited from as many potential sources as practicable. Indicate whether a FedBizOpps notice was or will be publicized as required by FAR Subpart 5.2 and, if not, which exception under FAR 5.202 applies. As required by FAR Subpart 5.2, a Notice of Intent (NOI) to award a sole source contract was publicized on beta.SAM.gov July 14, 2020 to announce the Government’s intent to obtain the BD FACSAria Fusion cell sorter on a non-competitive basis. Responsible sources who could provide a comparable product were provided an opportunity to respond with a capability statement. If a reasonable capability statement from another source was provided, the intent was to then post a Request for Quotations on a full and open competition basis. NIAID received no inquiries from the NOI.

8. Determination by the Contracting Officer that the anticipated cost/price to the Government will be fair and reasonable. The techniques identified at FAR 13.106-3(a)(2) will be used to ensure award will be at fair and reasonable prices. NIAID will consider the independent Government estimate, market research, comparison with similar items in the marketplace, and quoted prices found reasonable on previous purchases as determining factors for price reasonableness. A final determination by the Contracting Officer will be conducted prior to award of a purchase order.

9. Description of the market research conducted (see FAR Part 10) and the results, or a statement of the reasons market research was not conducted.

The LIR conducted a thorough search of manufacturer’s known for manufacturing cell sorting equipment. They found only one manufacturer, BD, that offers a cell sorter that appears to meet their specific requirements.

Although there are many types of cell sorters on the market, LIR found none that meets their immediate needs. All other sorters would require extensive training, a minimum of six months, with additional time to reconfigure sorting strategies and protocols before being able to conduct sorting of cells collected from rare and irreplaceable human specimens.

Similar cell sorters provided by other manufacturers are listed below. However, manufacturer information and product names are redacted not only to support the market research conducted, but also to ensure other vendors are not put to a disadvantage in future acquisitions.

1. Vendor A offers Model A with dimensions that may not fit in our limited space.

In addition, Model A is built with technology that has not been automated and upgraded. This means operators must perform many of the procedures manually that lengthens both the start-up and collection time each time the instrument is used, increasing the risk of low sorting yields and outcomes, in addition to increasing the need for routine laser, nozzle and stream alignment. Instrument laser alignment is a task normally performed by field engineers or experienced core operators. Any change in pressure or nozzle alignment during the sorting may also affect laser alignment and stability.

While LIR scientists are well trained, they do not possess these skills. The exporting of data with Model A also remains a challenge as it does not provide more direct interfacing between acquisition and analysis software.

Model A does not meet the following requirements, either by being absent or inferior:

Cell sorter must have capability of achieving high sorting rates, demonstrating linear and accurate data acquisition at above 50,000 events per second.

Cell sorter must offer user-friendly optical alignment, laser alignment or laser focus to reduce instrument set-up time and operator interaction in the biosafety cabinet.

Must have a gel-coupled cuvette that collects four times as much excitation light compared to stream-in-air systems to maximize sensitivity in signal detection (Flow Cytometry Lens System).

Cell sorter must not exceed dimensions of H 86.5 in, x W 50.2 in, x D

35.6 to fit in space available and biosafety cabinet.

Cell sorter must include an octagon shaped collection optics that increases the quality and quantity of the fluorescent signal. Fiber coupled from collection lens (Fluorescence Detection Instrument with Reflective Transfer Legs for Color Decimation).

Cell Sorter must self-regulate during sorting and have the ability to shut down sample flow, sheath flow, and cover sorted materials in presence of gross shift in sorting parameters (Flow Sorter with Video-Regulated Droplet Spacing).

A redundant aerosol management system must be included in event of blower motor failure. The system must evacuate the sort collection chamber by trapping aerosolized particles with a 0.10 m size ultra-low penetrating air filter.

Cell sorter must offer seamless integration with existing FACSAria Cell sorter for ease of assay transfer between sorters and minimum training of operators with identical sample acquisition software.

2. Vendor B offers Model B with lower sensitivity which means it is more likely to miss rare events. This is an important consideration given that there is a need to use the sorter to identify cells with rare properties, namely those that are specific for HIV and other pathogens, including influenza and the novel coronavirus that causes COVID-19 disease. The Vendor B instrument also does not have individual laser beams that are spatially separated to ensure maximal separation of populations stained with various fluorochromes in a pane. This limitation can reduce the ability to perform multiplex sorting, which is a new given the increasing number of populations identified in patient samples that need to be tracked separately. Finally, in addition to different instrument technologies, Vendor B uses special chips for sorting that are expensive and would need to be replaced upon clogging and for each sort.

Model B does not meet the following requirements, either by being absent or inferior:

Cell sorter must have capability of achieving high sorting rates, demonstrating linear and accurate data acquisition at above 50,000 events per second.

Cell sorter must offer user-friendly optical alignment, laser alignment or laser focus to reduce instrument set-up time and operator interaction in the biosafety cabinet.

Must have a gel-coupled cuvette that collects four times as much excitation light compared to stream-in-air systems to maximize sensitivity in signal detection (Flow Cytometry Lens System).

Sensitivity should have minimum resolution <118MESF on FITC and 34MESF on PE at 90KhZ and 70psi using Sphero rainbow beads.

Cell sorter must include an octagon shaped collection optics that increases the quality and quantity of the fluorescent signal. Fiber coupled from collection lens (Fluorescence Detection Instrument with Reflective Transfer Legs for Color Decimation).

Cell sorter must be able to utilize up to 6 solid state diode or DPSS lasers in various wavelengths and outputs lasers and up to 18 colors plus forward and side scatter.

Cell Sorter must utilize proprietary software that acquires and analyses in bi-exponential display format with FCS 2.0, 3.0 and 3.1 export of analyzed data.

Cell Sorter must self-regulate during sorting and have the ability to shut down sample flow, sheath flow, and cover sorted materials in presence of gross shift in sorting parameters (Flow Sorter with Video-Regulated Droplet Spacing).

A redundant aerosol management system must be included in event of blower motor failure. The system must evacuate the sort collection chamber by trapping aerosolized particles with a 0.10 m size ultra-low penetrating air filter.

Cell sorter must offer seamless integration with existing FACSAria training of operators with identical sample acquisition software.

Of the vendors considered, only BD Biosciences could meet the salient characteristics of the functionalities with regard to:

Cell sorter must have capability of achieving high sorting rates, demonstrating linear and accurate data acquisition at above 50,000 events per second.

Cell sorter must offer user-friendly optical alignment, laser alignment or laser focus to reduce instrument set-up time and operator interaction in the biosafety cabinet.

Must have a gel-coupled cuvette that collects four times as much excitation light compared to stream-in-air systems to maximize sensitivity in signal detection (Flow Cytometry Lens System).

Cell sorter must include an octagon shaped collection optics that increases the quality and quantity of the fluorescent signal. Fiber coupled from collection lens (Fluorescence Detection Instrument with Reflective Transfer Legs for Color Decimation).

Cell Sorter must self-regulate during sorting and have the ability to shut down sample flow, sheath flow, and cover sorted materials in presence of gross shift in sorting parameters (Flow Sorter with Video-Regulated Droplet Spacing).

A redundant aerosol management system must be included in event of blower motor failure. The system must evacuate the sort collection chamber by trapping aerosolized particles with a 0.10 m size ultra-low penetrating air filter.

Cell sorter must offer seamless integration with existing FACSAria training of operators with identical sample acquisition software.

All these salient characteristics are critical to maintain continuity of science and to support the advancement of research performed by LIR. The need for these features is not only to continue the use of current protocols and parameters without disruption and variability, but also to maintain the same extend of continuity in the future.

The other vendors cannot meet the salient characteristics of the specifications.