G
Required Features of High Sensitivity Laser Scanning Confocal
Microscope:
• Must incorporate a 34-Channel Quasar Detection unit, consisting of a calibrated 32-element Gallium Arsenide Phosphide (GaAsP) array PMT with typical QE >45 % (peak) and two calibrated flanking single PMT detectors; the one covering the long wavelength range is peltier cooled ideal for High Red Imaging dyes. This setup must enable image acquisition with freely definable emission windows and advanced spectral acquisition and analysis. The QUASAR detector array must enable three distinct modes of spectral image acquisition for ultra-sensitive imaging;
• Simultaneous Sensitive 34 detector spectral acquisition with 9.7nm spectral resolution (two side flanking PMTs in addition to the 32-element PMT array). Absolutely critical for highly sensitive spectral acquisition samples that require single scan light exposure and collection on a highly sensitive detector unit.
• Must include the 32-element linear GaAsP array PMT array featured in the QUASAR detection unit. This ultra-sensitive 32 channel detector must provide ultra-flexibility in the most sensitive applications. Must provide flexible imaging modes and unique ability to subdivide into up to 10 separate detectors in channel mode.
• Must include a microlens array is situated in front of the 32-element linear GaAsP array PMT to ensure there is no cross talk between detector channels and provide accurate spectral information.
• The 32-element linear GaAsP array PMT and actively cooled red enhanced PMT can be switched via software between 2 modes; optional software may be purchased to utilize the system in FCCS modes
• Digital oversampling imaging mode must allow users to control a high voltage gain and digital offset on the detection PMT.
• Photon counting imaging mode must allow for the system to distinguish the arrival of single photons many tens of times during pixel dwell time as an image is scanned (GaAsP PMTs and cooled PMTs are run at 15MHz). This mode must discriminate actual photon arrivals events to the detector and allow for samples with low amounts of fluorocromes (i.e. yeast) to be easily imaged and produce a noise free image.
• The highly efficient holographic grating must lead to an even spectral dispersion over the entire visual spectral range (380-750 nm). Grating utilizes a ‘spectral recycling loop' to recover non-separated light, redirecting it onto the holographic grating for optimum sensitivity. Linear light dispersion provided by a highly efficient grating is critical for applications where spectral unmixing is desired.
• Software integrated linear modulation of laser power from 0.001 to 100% for all laser lines between 458 and 633nm providing the ultimate sensitivity control to manage signal to the highly efficient 32 channel GaAsP detector unit. Reaching ultra-low laser powers particularly while imaging highly photo toxic or sensitive fluors can be critical.
Super Resolution Unique Features
• SR-SIM must provide a field-of-view of 80 x 80 μm in SR-SIM using a Plan-Apochromat 63x /1.4 Oil immersion objective lens utilizing an sCMOS camera. This must give users a large field of view for super resolution techniques which must be at least 2.56 times larger than when using a standard 512x512 pixel EMCCD. Many cells must be imaged in a single scan for high sup resolution throughput imaging.
• SR-SIM acquisition strategy must support both 3 and 5 grid rotations.
• Automatic (Transparent to the user) motorized switching between 5 different phase gratings (to change the grid spacing) must be present on the system. This must allow the use of multiple lenses and excitation wavelengths to achieve the theoretical resolution improvement for any fluorophore or sample.
• 4 Color Superesolution imaging of samples (labeled with 4 standard fluorophores or fluorescent proteins or any combination) must be possible with this system. Imaging Wavelengths must be included 405,488,561 and 640 and must be able to image all on a single sample. Standard Confocal preparations must be imaged in superesolution with this system.
General Features for the Confocal that are Required
• Must be able to freely rotatable scan field (0-360 degree in 0.1° steps) and free xy offset (crop functionality). Users require the flexibility to adjust the scan field without worrying how samples are mounted. Must allow easily flexible adjustment for highest speed imaging to align in X regardless of sample orientation.
• OSCiscan, Online Scanner Calibration, for artifact-free, fast and reliable bidirectional scanning without the need of manual adjustment. Users must be able to image with the point scanner with high frame rates reliably. Users must not have to worry about calibrating scanners for fast imaging modes regardless of image content. Fast point scan imaging is imperative for high quality resolution with high S/N content and easy transparent setup in a core facility is ideal.
• Gain calibration and linearization of up to 36 PMT detectors is foundation for reliable spectral acquisition and analysis.
• Must be able to upgrade with a 355 nm, solid state laser, 60 mW. Must be able to upgrade for future DNA damage experiments.
• Multiple (up to 99) freely definable real regions of interest (rROIs). Pixel precise control of the laser intensity and signal detection. Prevention of photo damage outside the rROIs boundaries necessary for work on living cells, FRAP, uncaging, photoactivation and photoconversion experiments.
• Individual AOTF settings (line selection and attenuation) per rROI.
• Use up to 8 laser lines simultaneously with full power possible from each laser line.
• System must offer at least 14 different scan speed levels (28 levels including bi-directional scans) for the wide variety of specimens and applications we will design. This is especially critical for NCI's single molecule experiments.
• Image format from 4x1 to 6144x6144 pixels in up to 35 detection channels simultaneously with up to 16 bit depth (65536 gray values) in all channels. (8, 12 OR 16 bit user selectable bit depth.)
• Automatic brightness compensation during the acquisition of z stacks by stepped increment of the excitation (laser intensity) and/or the detection gain (detector sensitivity). Must support at least a 5 point incrementation.
• Absolute linear scanner movement to ensure equal pixel dwell-times as a prerequisite for any quantitative studies.
• Spectral bandwidth for lambda stack acquisition must be selectable by the user. Must include a full spectral image that can be used for unmixing (suitable for spectral imaging of living cells and later unmixing without any spectral distortions).
• Reproducible high resolution spectral data acquisition with 3.2 nm or 4.9 nm steps through sequential spectral scanning in addition to the 10nm parallel spectral scanning.
• Optics must be designed for high transmission and aberration free imaging for 350 - 1100 nm.
• Pigtailed visible Lasers, for easily upgrades. Plug and play bayonette mounting for easy maintenance and reliable, reproducible operation. No mirror alignment required for lasers.
• Quantitative Colocalization must be part of the basic software with an interactive link between image display, scattergram and data table. The software must include interactive or automatic determination of thresholds, quantitative colocalization analysis with parameters like number of pixels, mean intensities +/-SD, colocalization coefficients, weighted colocalization coefficients, overlap coefficient after Manders, correlation coefficients, export of the analysis result and extraction of the colocalizing structures e.g. from image stacks.
• Transmitted detector must be included. Combination of a transmitted-light illuminator (e.g. a halogen lamp) with one of the detectors, fluorescent images can be superimposed with non-confocal transmitted light images.
Software Options:
• Quantitative Physiology package for acquisition of ion indicator dyes and display and analysis of ion concentrations including online and offline ratio for ratiometric dyes, online and offline F/F0 and DF/F for single wavelength dyes, calibration for single-wavelength and ratiometric dyes in situ and in vitro including background correction (after titration and with various curve fits or after Grynkiewicz).
• Multiple Time Series software package permits complex, combined time series with changing application configurations, autofocus and bleach functions.
• Tiling and Multiposition Scanning fully integrated into Software.
• Smart Setup Software for easy hardware control to adapt system configuration according to chosen dyes from a large database of fluorochromes. Different acquisition modes are suggested (e.g. Fastest and Best Signal).
• System must include self-test tool, with an easy to use maintenance user interface. In combination with a calibration objective. Allows automated pinhole adjustment and automatic scanner calibration, securing optimal system performance.
• Innovative concept for one touch button for reproducibility of experiments for multi-user environments via a REUSE function allows reactivation of all acquisition parameters necessary to reproduce an experiment (each image stores the entire method).
• Must be able to interchange parts, such as the PLAN APO objectives, if necessary.
• Must include at least a 12 months warranty on parts and labor.
• The new system must be compatible with currently owned Zeiss, and PC-based equipment.
• Must be able to network this new system to other computers and printers in the laboratory, thus allowing for easy image transfer and/or printing.
How to Submit a Response:
1. Page Limitations:
Interested qualified small business organizations should submit a tailored capability statement for this requirement not to exceed 10 single sided pages including all attachments, resumes, charts, etc. (single spaced, 12 point font minimum) that clearly details the ability to perform the requirements of the notice described above. All proprietary information should be marked as such. Responses should include a minimum of a two pages demonstrating experience over the past two years meeting the requirements of this notice. Statements should also include an indication of current certified small business status; this indication should be clearly marked on the first page of your capability statement (preferable placed under the eligible small business concern's name and address). Responses will be reviewed only by NIH personnel and will be held in a confidential manner.
2. Due Date:
Capability statements are due no later than 11:00 am. EST on August 1, 2014.
3. Delivery Point:
All information furnished must be in writing and must contain sufficient detail to allow the NCI to determine if it can meet the unique specifications described herein. Written responses can be emailed to Reyes Rodriguez Contract Specialist at [email protected] or mailed to the address located under Point of Contact. All questions must be in writing and can be faxed to (240) 276-5399 or emailed. A determination by the Government not to compete this requirement based upon responses to this notice is solely within the discretion of the Government. Information received will be considered solely for the purpose of determining whether to conduct a competitive procurement. In order to receive an award, contractors must have valid registration and certification in the Central Contractor Registration (CCR) and the Online Representations and Certifications Applications (ORCA), through sam.gov. No collect calls will be accepted. Please reference number SBSS-N02RC42593-57 on all correspondence.
Disclaimer and Important Notes:
This notice does not obligate the Government to award a contract or otherwise pay for the information provided in response. The Government reserves the right to use information provided by respondents for any purpose deemed necessary and legally appropriate. Any organization responding to this notice should ensure that its response is complete and sufficiently detailed to allow the Government to determine the organization's qualifications to perform the work. Respondents are advised that the Government is under no obligation to acknowledge receipt of the information received or provide feedback to respondents with respect to any information submitted. After a review of the responses received, an RFQ may be published on FedBizOpps. However, responses to this notice will not be considered adequate responses to a solicitation(s).
Point of Contact:
Inquiries concerning this Notice may be direct to:
Reyes Rodriguez
9609 Medical Center Dr, Room 1E128
Bethesda, MD 20892-9705
[email protected]