PD_Olympus_Electrophysiology_Rigs_FINAL.pdf

PURCHASE DESCRIPTION

1. Statement of Need and Purpose:

The unit on Neuromodulation and Synaptic Integration (NSI) is a new laboratory being established at the NIMH. This laboratory will utilize cutting-edge electrophysiological studies to determine how information is processed by the brain at the cellular, microcircuit, and systems levels to ultimately understand the computations that drive physiological behavior and psychiatric disorders. We utilize whole-cell slice electrophysiology in combination with other techniques, including optogenetics, UV uncaging, transgenic mice, viral approaches, and pharmacology to dissect how synapses and intrinsic factors that control excitability in neurons are regulated by receptors that are implicated in psychiatric disorders and are the major target of most pharmacological treatments for this disease. Electrophysiology is currently the only tool that allows neuroscientists to study synapses and intrinsic factors that control excitability with sub-millisecond temporal and single cell resolution. Moreover, with the addition of transgenic mice and viruses allow for the expression fluorescent proteins and optogenetic constructs in specific cell types to further dissect neuronal circuitry, it is imperative that the electrophysiological equipment used be equipped to image fluorescent proteins and photoactivate optogenetic actuators and UV-sensitive cage compounds. To meet the needs of the NSI Unit it has been determined that there is a critical need for the electrophysiology rig provided by Alliant/Olympus.

2. Background Information and Objective:

As the NSI will utilize whole-cell slice electrophysiology as a means to dissect the function of neuronal circuits implicated in psychiatric disorders, we seek to acquire two electrophysiology rigs with all the components necessary to successfully execute this cutting-edge technique.

Olympus has put together a comprehensive package for an electrophysiology rig that has all the features needed to successfully execute the mission of the NSI unit and the NIMH. The items being requested are BX51WI microscopes from Olympus equipped with cutting-edge imaging technology for improved imaging of brain slices, including infra-red Nomarski differential interference contrast (DIC) and a cooled, high-speed Retiga electro monochrome camera, as well as various dichroic mirrors for fluorescent imaging of various fluorophores. This system also offers the capacity to simultaneously do both DIC and fluorescent imaging. The system will also include a PE-300 LED system, which allows for imaging of fluorescent molecules during electrophysiological recordings, as well as optogenetic stimulation at various wavelengths, including UV stimulation for uncaging of photoactivatable compounds. One added advantage of the PE-300 LED system is that it can be controlled by transistor-to-transistor (TTL) logic by electrophysiology hardware/software, including control by a single input of multiple wavelengths at timed delays.

Moreover, this microscope will be equipped with Sutter Instruments MT-1000 support towers, which provide solid support of the sample and micromanipulators that house the microelectrodes used for recordings, as well as microstimulators used for stimulation. The MT-1000 support towers are also very versatile and lend themselves to the incorporation of up to four recording electrodes with independent support, which abrogates vibrations on separate electrodes. The MT-

1000 support towers also allow for easier disassembly of the system for routine cleaning and maintenance versus other conventional single platforms. Moreover, the system will come equipped with a MPC-285 micromanipulator bundle which provides accurate and precise control of microelectrodes and microstimulators. This system, unlike many of its counterparts, allows up to 4 independent micromanipulators to be controlled by a single Rotorary Optical Encoder (ROE) controller and provides up to 10 different speeds for versatile movement velocities. This bundle also includes a chamber platform, low-noise imaging chamber, and heating element system that provides a stable sample and maintains accurate and precise control of the temperature in the slice preparation. This bundle includes a TMC Cleanbench airtable to isolate vibrations and provide stability in the brain slice preparation and of the micromanipulators. This table is essential to ensure quality electrophsiological recordings and data. Last, but not least, purchase of this equipment comes with tech support from trained Olympus technicians who will come on-site if needed to cater to any technical issues that might, and likely will, arise.

Taken together, Alliant HealthCare Products through their partnership with Olympus has put together a comprehensive package for an electrophysiology rig that has all the cutting-edge components needed for the NSI unit. This rig will provide all the technology necessary to acquire the cutting-edge data that will help us understand brain function in physiological conditions and dysfunction in psychiatric disorders. The addition of this system will greatly enhance the NIMH mission statement as well as the NSI Unit.

3. Generic Name of Product: Microscope, table, and associated accessories for whole- cell slice electrophysiology studies

4. Purchase Description: Brand-name or Equal

Two Olympus electrophysiology rig to include:

1. BX51WI microscope with associated components, quantity 2

2. Retiga Electro Camera and Software, quantity 2

3. Sutter MT-1000 with MPC-385 micromanipulator bundle for BX51WI microscope, quantity 2

4. TMC Cleanbench air table and accessories, quantity 2

5. HP-G4 Z4 Tower Computer, Win10, Intel Xeon W-2102 2.9 2400MHz 8.25 4Core CPU, 32GB

DDR4-2666 RAM, 512GB SATA SSD, 2x1TB SATA HDD, Quadro P600 2GB, quantity 2

6. Warner Instruments PH-1 chamber platform, low-noise imaging chamber, TC-344F Heater

Controller, SF-28 slow flow solution in-line heater, chamber heater, CC-28 cable assembly for heater controllers, quantity 2

5. Salient characteristics:

1. The product should have visible light DIC to allow for operator high-resolution observation of the surface of the tissue.

2. The rig should have a combination of an IR camera and 775 nm IR-DIC observation accessories.

3. For deeper observation into the tissue the device should have a polarizer and analyzer for 900 mm Nomarski DIC.

4. Must have a condenser with a high numerical aperture (NA), short working distance condenser to offer improved contrast suitable for use in visible and 775 nm to 900 nm near-infrared light.

5. Must have an oblique condenser with a long working distance enables the angles of shadow to be altered through 360 degrees without moving the specimen. This must require no additional accessories, and the oblique illumination should be easy to set up and control. The oblique illumination slit aperture should be variable in size and on a slider allowing quick changeover.

6. The objectives of the microscope must have a long working distance and be dipping objectives designed with special angles for experiments in electrophysiology.

7. The objectives must have a water immersion cap to remove image aberrations caused by ripples on the water surface of immersed specimens.

8. The setup must come with a fast, high-quality cooled CCD monochrome camera with low electrical noise suitable for electrophysiological experiments. Moreover, the camera must have software for acquiring, viewing, and manipulating images that can be run concurrently with electrophysiological hardware and software.

9. The swing nosepiece holding the objective must have a unique slim, compact design and front-to-back swing motion to enable objective changes without interfering with electrodes and micromanipulators. This must also incorporate objective positioning with a vibration-free counter spring mechanism to prevent mechanical distortion of the brain slice and electropdes.

10. The microscope must have a LED or laser light source for simultaneous and/or independent photoactivation of optogenetic constructs with various wavelengths, as well as the capability for UV uncaging of photoactivatable compounds.

11. The LED or laser light source must also be able to be used for imaging of fluorescent molecules in addition to serving as a light source for optogenetic and UV uncaging experiments.

12. The LED and laser light source must be TTL-triggered, able to deliver a TTL to the digitizer of the electrophysiology rig, and be programmable to deliver different duration and/or wavelength pulses of light to the specimen.

13. Must have support towers instead of bulky single platform to allow for the recording multiple neurons and easy disassembly and maintenance, and reduction of vibrations by multiple manipulators on the rig.

14. Support towers and chambers must be compatible with parts from multiple vendors.

15. The electrophysiology rig must include an in-line heater, a heating element for simulatenous temperature control in the slice bath, and a heating controller that allows for control and monitoring of temperatures with variable speeds in both the bath heating element and in-line heater.

16. The equipment must have a chamber platform that accommodates the slice bath chamber and connects with the heating element to heat the slice bath chamber.

17. The micromanipulator controller must be able to accommodate at least four micromanipulators and have variable speeds, as well as the ability to bring it to a home position and working position.

18. Micromanipulators must be compatible with commercially-available microelectrode and microstimulator holders from various vendors.

19. The micromanipulator must turn “off” electronically when not in use to minimize electrode drift and electrical noise.

20. The airtable must provide vibration isolation, in line with the quality that only TMC airtables provide.

21. Must come with Instrument support from field technicians

6. Quantity:

Item Number Description Quantity

Olympus electrophysiology rigs and associated accessories as specified in this purchase description

7. Delivery:

Delivery of all items shall be within thirty (30) days after receipt of order to the following address:

35A Convent Drive, Room 2E461, Bethesda, MD 20892

Attn: Dr. Hugo Tejeda

All deliverables shall be packaged and shipped in accordance with Government specifications. At a minimum, all deliverables shall be marked with the contract number and Contractor name.