National Institutes of Health, National Heart, Lung, and Blood Institute, Contracts Operations Branch, 6701 Rockledge Drive, Room 6100, MSC 7902, Bethesda, MD 20892

66 -- CONFOCAL LASER-SCANNING INSTRUMENT FOR STUDY OF METABOLISM OF INTACT CARDIAC MYOCYTES SOL NHLBI-HL-P-98-260 POC Gina M. Cianflone, Contract Specialist, 301-435-0363 This is a synopsis for commercial items, prepared in accordance with the format in FAR subpart 12.6, as supplemented with additional information included in this notice. Proposals are being requested and a written solicitation will be issued. The Laboratory of Cardiac Energetics, Division of Intramural Research, National Heart, Lung, and Blood Institute (LCE, DIR, NHLBI) of the National Institutes of Health (NIH) has a requirement to obtain one (1) Confocal Laser-Scanning Instrument for Study of Metabolism of Intact Cardiac Myocytes. Specific Requirements: Specifically, the contractor shall: 1) Construct a CLSM system capable of state of the art imaging of single cells. 2) Install the system into the appropriate area at the NIH. 3) Train the LCE staff in any special hardware or software required for the routine operation and maintenance of the system. The confocal laser-scanning instrument described above shall be capable of accommodating multiple users and have the capacity for future upgrading as theneed arises. It shall be a versatile system optimized for ease of use and maintenance. Specifically, it shall include the following features: 1. Basic System, Lasers, and Filter Sets: a)The basic system shall be compatible or adaptable to accommodate present equipment including the LCE's current Zeiss inverted microscope (model # 135M) and CCD camera (Hamamatsu XC-77); b) The system shall be compatible with current objectives on the LCE's inverted microscope (Zeiss Fluar 10, 40, and 100x, and Zeiss C-Apochromat 63x objectives); c) The system shall have a motorized stage which can be software controlled to allow for z-sectioning to a precision of 1 m; d) The basic system shall have ultraviolet, argon and helium/neon lasers. Scanhead filter and dichroic mirrors sets shall include DAPI, Rodamin, Fluorescein, Snarf, CY2, CY3, and Lucifer Yellow. These lasers and filter sets will enable imaging of NADH autofluorescence and other energetic parameters in isolated cardiac myocytes including mitochondrial membrane potential, intracellular pH, and various intracellular ions; e) The system shall also allow for transmitted light imaging; f) The system shall have a minimum of three photomultiplier tubes to allow for simultaneous multi-wavelength signal collection. Multiple photomultiplier channels will allow researchers to examine several different physiological parameters at the same time; g) The system shall allow for attenuation of laser intensity over a broad range using either Acousto Optical Tunable Filters (AOTFs) or neutral density filters. These filters shall allow attenuation from 0-100% in 1% increment; h) The system shall include appropriate laser cooling apparatus and an anti-vibration table for the microscope and scan unit. 2. Automation, Scan Control and Automatic Hardware Optimization: a) Control of all operations including changing filter sets, pinhole settings, laser attenuation, laser excitation wavelength, or photomultiplier channel shall be automated in the scan head to allow for efficient and fast movement between experimental configurations; b) Scan control shall include a zoom feature and shall be digital to allow optimal scanning specificity; c) The system shall have feedback sensor for evaluating and adjusting laser intensity to minimize laser amplitude fluctuations. 3. Image Quality: The offerors shall submit UV NADH fluorescence images for assessment of image quality. Images from vertebrate cardiac myocytes or skeletal muscle cells are desirable but images from other living cells will be acceptable . If living cells cannot be imaged, offerors shall submit an image of a sample of 1 M NADH (reduced form of nicotinamide dinucleotide) in a container of the offerors choosing and provide the rate of photobleaching of the NADH autofluorescence signal (366 nm UV excitation, 455 nm emission) at laser powers attenuated to below 10%. Images demonstrating the best resolution provided by their system using objectives with N.A.s of at least 1.2 and magnification of 63-100x. Submitted images can be of any samples though living cells are preferable. 4. Image: a) Image size shall be at least 512 X 512 pixels but preference would be given to systems with image capabilities of 1K X 1K; b) Imaging resolution shall approach the submicron level at optimal experimental configurations; c) Imaging speed shall be a minimum of 2-4 frames/sec for 512 X 512 image sizes; d) Adequate handling of samples with wide range of intensities requires that the system provide image depth of 8-12 bit digitization. 5. Computer Hardware and Software: a) The system shall include a state of the art computer which controls all aspects of image acquisition, microscope operation, and image processing. These requirements necessitate at least 128 Mb of Ram and 8 Mb of VRAM, SCSI interfaces to CLSM hardware, large hard drive memory (at least 3 Gb), and at least 1 large video monitor to accommodate multi-image display; b) Software control shall include frame averaging, various on-line filtering capabilities, ratioing methods, and user specified experimentconfiguration. At the very least the user specified experiment configuration shall provide options for automating sequential image acquisition (time series physiological studies) and storage of acquired images, z sectioning and volumetric reconstruction. The award of a firm, fixed-price type contract is anticipated. This requirement is not a set aside. The applicable Standard Industrial Classification (SIC) is 3821, size standard 500 employees. The proposals may be due fifteen calendar days from the publication date of this synopsis. Interested parties should submit a written request (via mail or facsimile transmission to (301) 480-3345) for a copy of the solicitation number NHLBI-HL-P-98-260 to the National Heart, Lung, and Blood Institute, Contracts Operations Branch, Procurement Section, Building RKL2, Room 6150A, 6701 Rockledge Drive, Bethesda, Maryland 20892-7902, Attention: Gina M. Cianflone. All responsible sources may submit a proposal, which if timely received shall be considered by the agency. (0097)

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