D JA_Redacted.pdf

Justification and Approval for Sole Source

“Source Selection Information – see FAR 2.101 and 3.104”

1. Identification of the agency and contracting activity.

a. Federal agency and contracting activity.

Department of Health and Human Services National Institute of Health

National Institute on Drug Abuse

b. Sponsoring organization.

NINDS, Division of Intramural Research, Stroke Branch, Neurovascular Research Unit.

c. Project Officer information.

• E-mail Address.

2. Nature and/or description of the action being approved.

a. Acquisition purpose and objectives.

The study of vascular function is central to understanding neurovascular biology. We need in vivo Imaging approaches to simultaneously study the structure and functions of neurovascular coupling units (endothelial cells, mural cells, neurons and glial cells). Our study will greatly benefit from using a multiphoton microscope to image and manipulate specific neurovascular cell populations using 3D spatial light modulator (SLM) optical stimulation in awake and behaving animals. This will facilitate our research to achieve a more comprehensive understanding of neurovascular mechanisms in regulation of blood flow in the physiology and disease models of vascular cognitive impairment and dementia (VCID), including stroke.

One of the aims in the Neurovascular Research Unit is to evaluate structures and functions of cerebral vasculature in vivo. In order to do conduct these experiments, we heavily rely on a two-photon microscope system which allow us to monitor different compartments of vasculature (arterioles, capillaries and venules) along with neurons and glial cells with high spatiotemporal resolution while optically manipulating specific cell populations.

b. Project background.

Madison, WI

b. Nature of the acquisition and proposed unique qualifications of the contractor(s).

Based on market research that was conducted with vendors who provide multi-photon microscopy systems with 3D spatial light modulator (SLM) photostimulation, it was determined that Bruker Nano, Inc is the manufacturer of the product that best meets the needs of the Neurovascular Research Unit. Additionally, it has been confirmed by the manufacturer that the equipment is only available direct. Finally, the CO confirmed that the equipment is not available via any GWACs.

Specifically, the following technical specifications of the Bruker Nano system make it uniquely able to meet the need for in vivo imaging approaches to simultaneously study the structure and functions of neurovascular coupling units (endothelial cells, mural cells, neurons and glial cells).

• Scanning Field-of-View: The 2P Plus scan head realizes a new optical design optimized for extended scanning field-of-view in galvo mode.

The system maintains PSF within 1 axial PSF across entire field at scan zoom 1x. Overscan allows for scan zoom 0.8x for 1.4mm x 1.4mm field of view with Nikon 16x 0.8 objective. With an overscan Field Number of 28.3mm the 2P Plus has the largest scanning field of view of any commercial system.

• Close Proximity Detectors: The 2P Plus offers up to four detectors that are in close proximity to the objective and move in unison with the objective. The short and fixed path length between the collection lens and objective allows for more efficient collection of scattered photons and 14° angle of collection from back aperture of the objective.

• 2” Collection optics: The 2P Plus uses 2” collection optics from the objective to the detectors to ensure that the diverging photons end up hitting the photo cathode of the detectors instead of getting lost along the way.

• Rotating Nosepiece: Single axis rotating nosepiece that swivels 90 degrees on either side of its normal position. This allows users to image regions that are on the back or the side of the head such as auditory cortex. This feature does not require any re-orientation of the scan head or realignment of laser transfer optics, nor does it affect the optical relationship between the objective and PMT photo cathodes.

• Resonant Scanner “add-on”: The optional resonant scanner in the 2P

Plus scanhead is a true add-on, meaning it is relayed onto the standard 6mm imaging galvos. This architecture allows for either traditional raster scanning in galvo mode (adjustable dwell time, line scans, ROIs, etc.) or high-speed scanning in resonant mode, selectable via drop-down in software for seamless switching.

Resonant ROIs have increased SNR as the resonant scanner can be re-centered in the field of view with the x-galvo. The resonant scanner can be installed as a field upgrade.

• Photoactivation: The 2P Plus can be upgraded with full field

Photoactivation with LEDs. The system includes inputs and glass for two LEDs, designed for 470nm and 590nm. Photoactivation can be performed rapidly during imaging when high speed shutters are used.

This allows the users to do optical stimulation experiments with minimal delay between activation and imaging.

• Neuralight 3D Ultra SLM: 2P Plus can be upgraded to perform simultaneous multi-point optical stimulation/activation in the secondary galvo pathway. Utilizes a custom Spatial Light Modulator for dynamic control of the wavefront in Fourier space to manipulate the number of spots created and their position in 3D space. Coupling into the galvo path allows for use of diffraction-limited spots that can be spiraled rapidly with the galvos, maintaining photon density for reliable activation, and covering larger areas (e.g. cell bodies) with the spirals.

This enables the Bruker SLM module to take advantage of existing uncaging, raster, point, and spiral activation protocols that have been vetted by the scientific community.

15.4 x 15.4 mm (768 x 768 pixel) square chip to accommodate a large circular 15.4 mm beam

1.7 ms hologram to hologram response time (not just electronics controller response time) Field of View 700um x 700um x 700 um field of view with 16x 0.8

NA objective 10um Z PSF 10us phase mask timing jitter Built-in wavelength calibration optical path to optimize SLM efficiency for actual laser on system and also for a different wavelength

Fully integrated into Prairie View software for GUI-based Mark Points experiments in 3D

True simultaneous volumetric imaging during photostimulation with decoupled ETL module

Can be used to generate traditional holographic patterns on the sample

• Moving Microscope: Translation along X and Y axes, up to 1.5”, in addition to the standard Z axis 1.25”. This allows use of simple, non-motorized, experiment-specific stages for in vivo experiments where the specimen is held stationary while the scope is moved around (such as a head-fixed mouse running on a ball or a customized tread mill). This also enables Atlas Imaging, a software feature for visualizing a montage and picking out ROIs from a large sample area.

• Easy access to animal/specimen for behavioral stimulation: The microscope has been specifically designed for in vivo work. Its slender design allows plenty of room underneath the objective to accommodate diverse experimental needs. In addition, the design also allows plenty of room around the scope to bring in behavioral equipment close to the animal without spatial constraints. The 2p Plus also allows more room between the objective and the scope body to accommodate cases where the animal’s orientation has to be changed for behavioral stimulation.

• High Efficiency GaAsP detectors with shutters: The detectors are cost-effectively upgraded to high sensitivity GaAsP detectors hand-selected for QE > 45%. These detectors have about twice the QE of multi alkali detectors. High-speed shutters are integrated into the detector mounts to protect the PMTs from exposure to any Photoactivation light or ambient light when not in use for imaging.

• More than 1000 frames per second imaging: Resonant scanner enables high speed scanning (30fps at 512 x 512 resolutions at full field of view). This is particularly useful for Calcium imaging. Use of ROIs (regions of interest) with resonant scanner provides even higher frame rates limited only by the number of lines in the ROI. These ROIs can have any number of pixels along a line leading to better SNR, and better sample viability. Since the ROIs are not limited to 32 or more lines, frame rates greater than 1000 fps can be achieved; for instance 790fps for 512 x 16 ROI or 1319fps for 512 x 8 ROI.

Software:

• Tiling software for Whole Brain Imaging: Software provides a highly intuitive user interface to capture high resolution images of large samples (whole brains in 3D). The preview mode allows users to setup the acquisition, and the software combined with high precision stages collects the images in “tiles” to construct the whole image in 3D. This feature is enabled by the motorized XYZ microscope and is fully functional on sample stages with static position.

• User login based software configuration: The software supports multiple users and saves their configurations within their logins. It also allows users to save multiple configurations (relating to different experiments) for fast and easy re-loading of correct experimental parameters.

• Real Time Data Analysis and Feedback: The software makes the data available to an external program so that the data can be analyzed in real time if required. For instance, a third party program such as Matlab can request image data from the software, analyze the image and send commands back to the software to move the stage to bring the cell of interest back to the middle of field of view. All of this can be done in real time.

• Synchronization with ePhys: The software provides accurate timing information about scans using pixel clock, line clock, and frame clock signals (TTL). These signals can be used to trigger external devices.

In addition, the software provides the ability to produce up to 8 analog outputs and record from up to 16 analog inputs synchronized with photo-stimulation experiments.

5. 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.

A Non-competitive Combined Synopsis/Solicitation will be posted to SAM.Gov.

6. Determination by the Contracting Officer that the anticipated cost/price to the Government will be fair and reasonable.

The CS will conduct a price analysis to determine that pricing obtained is fair and reasonable, to include a comparison of any quotes received, comparison to the IGCE, and comparison to previous purchases of similar equipment.

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

Exploratory discussions with vendors who offer this product were held in Oct- Dec 2023. In addition, discussions were held with Investigators in NINDS and NIMH as they have purchased the same laser scanners from Bruker Nano.

Additionally, the CS conducted market research to determine the extent that the required equipment is available. Market research included:

• Searching Interagency Contract Directory

• Searching GSA eLibrary

• Searched FPDS

• Searched SBA’s Dynamic Small Business Search

• Contacted manufacturer to determine available sales channels

• Searched internet - Google.com

8. Any other facts supporting the use of other than full and open competition.

Not applicable.

9. Listing of sources, if any, that expressed, in writing, an interest in the acquisition.

Vendors that expressed interest after exploratory discussions were: Bruker Nano, Inc., Nikon and Thorlabs. However, based on these discussions and/or demos, it has been determined that the Ultima 2Pplus Laser Scanner; multi-photon microscopy system with 3D spatial light modulator (SLM) photostimulation from Bruker Nano uniquely fits the needs for this procurement.

Statement of the actions, if any, the agency may take to remove or overcome any barriers to competition before any subsequent acquisition for the required supplies or services.