D Purchase Description_Final.pdf

Version 11-30-2016-1 FAR Part 11—Describing Agency Needs

PURCHASE DESCRIPTION

1. Statement of Need and Purpose:

The study of vascular function is central to understand the 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.

2. Background Information and Objective:

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.

3. Generic Name of Product: Multi-photon microscopy system with 3D spatial light modulator (SLM) photostimulation

4. Purchase Description:

Bruker Nano, Inc. Ultima 2Pplus Laser Scanner (Brand Name Only); multi-photon microscopy system with 3D spatial light modulator (SLM) photostimulation, and accessories, installation, and warranty.

5. Salient characteristics

1. Scanning Field-of-View: The scan head realizes a new optical design optimized for extended scanning field-of-view in both galvo mode and resonant 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.

2. 3D Volume imaging with ETL: The microscope includes a motorized Z focus; it also includes an electrically tunable lens (ETL) in the resonant scan path. The ETL provides high-speed dynamic positioning of the focal plane for Z-stacks. It is also decoupled from the photoactivation scan path, enabling simultaneous imaging and photoactivation from different Z-locations.

a. 480µm range

b. 15ms step response, can be driven faster depending with tradeoff in ringing

c. Magnification corrected at each limit of range to 1.1x

d. Spherical aberration corrected with custom aspheric lens

e. Axial PSF measured 5 – 5.5µm throughout range

3. Close Proximity Detectors: it must offer 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.

4. 2” Collection optics: it 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.

5. Synchronization via BNC connection for timing of experimental apparatus is to be standard.

6. 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.

7. Resonant Scanner “add-on”: The resonant scanner 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.

8. Optogenetics: It includes a secondary galvo path for stimulation using a separate laser (Coherent Monaco: 1040 nm). This pathway is independent from the imaging pathway, enabling simultaneous imaging and stimulation through the Mark Points software module.

Allows for synchronization with analog voltage input/output and imaging acquisition.

9. 3D SLM: 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 SLM module to take advantage of existing uncaging, raster, point, and spiral activation protocols that have been vetted by the scientific community.

a. 512 x 512 pixels

b. 7ms response time using reflective LCD surface

c. Numerical Aperture selectable via lens sets, 0.6 or 0.8

d. Field of View 150 x 150 x 150um with 0.8 NA or 300 x 300 x 300um with 0.6NA

e. Wavelength range 660 to 1380 nm

f. Optional Dielectric mirror for increased reflectivity with tradeoff in wavelength range and rolloff

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

h. Can be used to generate traditional holographic patterns on the sample

10. 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.

11. 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.

12. 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.

12. Timing and Recombination Unit for timing synchronized beams, application for multi-color two-photon fluorescence microscopy

13. Fixed dispersion compensation for 1045 nm beam, 15000 fs^2

14. 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.

15. 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.

16. 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.

17. 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.

6. Quantity: Quantities and descriptions as follows:

Model Qty Description 2PP-01 1 Ultima 2Pplus Laser Scanner 2PP-MB 1 Upgrade to Moving X-Y Platform for Ultima 2Pplus 2PP-RS 1 Resonant Scanner High-Speed Optics Set for 2Pplus

2PP-U6-

1040 1 Uncaging Optics and Functional Mapping System for2Pplus (1040nm) - 6mm 2PP-SLMU 1 NeuraLight 3D Ultra SLM Module for PhotoactivationPathway 2PP-Z-ETL 1 Decoupled ETL Remote Focusing Module for Ultima 2PplusResonant Scanner v2.0

HED-UP-2 1

Upgrade of Both Multi-Alkali PMTs to GaAsP for High-Efficiency Detector Module (0MA and 2 GaAsP Total)

HED-HSS 2 High-Speed Shutter for H10770 PMTs

HED-

10770-mount 2

High-Speed Shutter Mount for Hamamatsu 10770 PMT -High-Efficiency Detection Module

518708 1 High-Speed Shutter Controller 518066 1 Epi-Fluorescence Filter Set: ET-DSRed (TRITC/CY3) for Nikon 516563 1 Nikon OBJ CFI75 16X/0.80 WD=3mm U-7885 1 Beam Expander Lens Set 521323 1 Tabletop dichroic, 1040dcrb LI-AOM 2 Laser Input Optics - Imaging/Uncaging with IntegratedAOM Install-17 1 Installation and Training - Dual Laser Input

EXWARR-

2PP 1 Extended Warranty - Ultima 2Pplus

7. Delivery Date: Expected delivery date is 150 days after receipt of order. Delivery must be made to Dr. Amreen Mughal, Porter Neuroscience Building, Building 35, 35 Convent Drive, Room 3D734, Bethesda, MD 20892-00001. No drop delivery allowed.

8. Period of Performance: Extended Warranty - One year extension of new system warranty of customer's system.

9. Option(s): N/A