Attachment 1 - PD_CO review.pdf

PURCHASE DESCRIPTION

1. Statement of Need and Purpose

The Laboratory of Circuits, Synapses, and Molecular Signaling (LCSMS) requires a complete system for brain slice electrophysiology that is compatible with its existing systems

(Scientifica SliceScope Pro 6000).

2. Background Information and Objective

The LCSMS studies fundamental issues of synaptic transmission, neuronal excitability, neuromodulation, and neural circuit activity using animal models (principally, the mouse). A major line of research employs patch clamp electrophysiology, fluorescence imaging, and optogenetic stimulation in brain slices. For this research, the LCSMS requires an electrophysiology rig capable of two (2) simultaneous patch clamp recordings from cell bodies and dendritic structures in mouse brain slices, imaging of fluorescently tagged neurons and dendrites, and widefield photoactivation with an LED (i.e., optogenetic stimulation or neurotransmitter uncaging). The rig must be compatible with the existing electrophysiology rigs of the LCSMS, in both hardware and software. These existing rigs are built around the Scientifica SliceScope Pro 6000 system. The components of the new rig must be interchangeable with components of this system and must run the same software.

3. Generic Name of Product

Brain slice electrophysiology rig.

4. Salient Characteristics

The LCSMS requires a patch clamp electrophysiology rig that will allow component swapping with its existing systems and that runs the same software. The existing systems are built around the Scientifica SliceScope Pro 6000 with Patchstar manipulators. (Note: an air table, a Faraday cage, and patch clamp amplifiers are not required, as the LCSMS possesses this equipment already.)

The detailed specifications are as follows.

A. Microscope, staging, software and optics

1. The offer must include a large, motorized horseshoe-size base plate (≥ 575 mm in width) to hold all manipulators and devices next to the sample. This stage must include 3 mounting carriages to ensure easy placement of micromanipulators.

Attachment 1 - Purchase Description

2. The offer must include cross-roller bearings in the motorized base plate stage to provide ultra-smooth and vibration free movement, with an overall range of ≥ 50 mm.

This must be controlled with a Patchpad device.

3. The staging must include a magnetic mounting bath perfusion chamber to hold in vitro samples. This must be mounted on a post and platform device on the stage, so that one can alter the height of the platform according to the microscope objective.

This must also include options to hold 35 mm petri-dish samples and clips for slide mounting.

4. The microscope must be fitted with an ultra-smooth motorized Z-axis drive that ensures vibration-free movement over 25 mm; this must be controlled with the same

Patchpad device as the microscope Z axis.

5. The microscope must have a small footprint and be very thin for easy access to the sample; no wider than 150 mm in width and 260 mm in length.

6. The microscope must be able hold Industry leading optics (Olympus or Nikon) to produce images.

7. The microscope must utilize a range of different contrast techniques to allow versatility between techniques. In this instance a ‘oblique contrast’ configuration is required. This oblique condenser must be an achromatic/aplanatic condenser, with numerical aperture of 0.8 and a long working distance of 5.7 mm.

8. The microscope must contain a 4x and 40x objective with a ‘swing-nosepiece’ to change between the 2 magnifications. The 4x must come with a working distance of

18.5 mm and numerical aperture of 0.8. The 40x must have a long working distance of 3.3 mm, a numerical aperture of 0.8, and should be compatible with Ultraviolet to

Infra-red spectrum transmission.

9. The microscope must come alongside with a complementary software to control all the motorized components including the micromanipulators/Microscope Z control and

XY staging.

10. The motors in the microscope must hold an infinite number of memory positions which allow repeatable positions for returning to sites of interest via the complementary software.

11. The microscope must include a 780 nm transmitted LED for far Infra-red sample analysis.

12. The microscope must include a C-mount adapter for the camera (with specifications of the camera below) alongside a single port for infrared applications.

13. The software must include free unlimited support and upgrades for lifetime use.

14. The software must have a virtual joystick to control the motorized components.

15. There must be an option to switch between a pad device and a cube device without any alteration of any other components – ‘plug and play’.

B. Optogenetic filter sets for full field stimulation: (cleanup/long pass filters)

1. The system must include the correct filters for full field optogenetic stimulation/silencing of 535 nm and 470 nm wavelengths using the fluorescence turret and a fluorescent LED.

2. The 470 nm clean-up filter must be installed in the LED.

3. The 535 nm clean-up filter must be installed in the LED.

4. The dichroic mirror must be situated in the fluorescence turret and allow simultaneous LED stimulation/silencing.

C. Camera

1. The camera must be a CCD camera with at least 24 fps at full resolution (at least 40 fps binned 2x2) and at least 14-bit resolution. It must include a fast USB 3.0 PCIe interface alongside software. It must be grounded (to prevent electrical noise), heat-sinked (instead of fan to prevent vibration) and sensor-cooled (for chronic electrophysiology experimentation).

D. Motorized Manipulators

1. Manipulators must include superior roller bearings for smooth movements.

2. Manipulators must have a theoretical resolution of 20 nm.

3. Manipulators must be controlled via a control cube device.

4. Manipulators must have a circular rotatory base for easy pipette exchange, no larger than 110 mm in diameter.

5. The manipulators must have exact matching axis modules, so they can easily be reconfigured and exchanged if one axis were to fail.

6. The manipulators must have a step in/out function to allow users to move along in a certain axis at a defined speed/distance; this must also be included on the control device.

7. The manipulators must include a specific bracket to allow a steep approach to the sample.

8. The manipulators must have a sliding head-stage for easy pipette exchange.

9. Manipulators must be able to control the speed from 0.1 microns/second to 4 mm/second

10. Manipulators must have a drift below 1 micron every 2 hours.

11. Manipulators must be compatible with a headstage compatible with the Axopatch

200B (Molecular Devices).

12. Manipulators must come with a ‘follow control’ software function that interlinks it with staging to ensure pipettes remain in field of view when panning around the sample.

E. Fluorescence

1. The microscope must include fluorescence turret beam path that has been completely optimized with fluorescence transmission/color correction and homogeneity. This must be able to attach onto the top of the upright motorized microscope.

2. The turret must include space for up to 6 fluorescence filter cubes for future compatibility with different fluorescent dyes. These cubes should be easily exchanged and moved by removing the front of the turret.

3. The fluorescence turret must be ‘push and click’ for quick change of filter cubes during experimentation.

4. The fluorescence turret must include filter cubes for GFP (green fluorescence protein) and mCherry/Texas Red dyes.

5. The filter cubes must contain emission, excitation and dichroic mirrors; to permit the correct light spectrum to reach the sample and the visualization of that fluorescence in that sample via the specified camera. Filter cube spectrum specifications must include: For GFP: 470/40x, 495LP, 525/50m, for mCherry: 560/40x, 585LP, 630/75m.

6. The fluorescence turret must include a space for a fluorescence light source.

7. The light source should be a white LED with TTL trigger control for the individual red, blue and green LEDs. This unit must include a liquid light guide for attachment onto the fluorescence turret - it must not be a direct fit LED as this can lead to possible sample vibration through the LED cooling fan. The LED system must have a USB interface to be controlled by a computer.

F. Bath saline perfusion

1. The system must a peristaltic pump and low-noise temperature controller for perfusion of bath salines.

2. The pump must have manual variable speed (1-100 rpm in 1 rpm increments), speed scrolling, priming, and a keypad lock. It should be comprised of a twin channel pump head and accept 0.8 mm wall, three-bridge manifold tubing in sizes from 0.13 to 2.79 mm internal diameter. Flow rates of 0.0001 to 36 mL/min at 30 psi should be possible.

3. The temperature controller must include a Peltier pre-heater block with a built-in temperature sensor and a bath sensor.

5. Quantity

One (1) system

6. Delivery Date

The Contractor shall deliver and install the equipment within 90 days after receipt of order.

Delivery must be Freight on Board (FOB) Destination and include inside delivery. On-site training must take place within two (2) weeks of equipment installation. The equipment shall be delivered and installed in coordination with the Contracting Officer’s Representative

(COR). The contractor shall provide an Original Equipment Manufacturer (OEM) certified service technician to perform setup, installation, and an on-site training session for staff members.

7. Warranty

The Contractor shall warrant that that the Equipment will be free from defects for a period of twenty-four (24) months from the date of installation, inspection by the Government and acceptance.

The Contractor must have a dedicated service team. This service team should be able to exchange faulty items with their service stock within a week of the user’s equipment fault.

The Contractor must provide an installation & training day for at least 4 users, installing the system fully and training the users on the system. This will include:

• Unpacking, assembly and full build of the manufactured products and produced software.

• Installation of the optical components, cameras and light sources onto the microscope and alignment where suitable.

• Full product training on all products as part of the installation.

• An overview of the operation of the entire system and the supplied software.

• Guidance and advice on the installation of third party (non-manufactured) products supplied by the Contractor.

8. Delivery Requirements

The Contractor shall deliver and install the required equipment in coordination with the COR.

Delivery must be FOB Destination and must include inside installation. This system will be delivered to laboratory space in Building 35A. Coordination of delivery will be performed with the COR at the time the equipment is ready to be shipped. The equipment shall be delivered and installed between the hours of 8:00am and 5:00pm, Bethesda, MD local prevailing time, Monday through Friday.

Laboratory of Circuits, Synapses, and Molecular Signaling (LCSMS)

National Institute of Neurological Disorders and Stroke

Building 35A/ Rm 3B-1008

35A Convent Dr

Bethesda, MD 20892