33565-3A-ACAN - STOCHASTIC OPTICAL RECONSTRUCTION MICROSCOPY (STORM)

BACKGROUND:
 
Notice is hereby given by the University of Ottawa of the intent to purchase from Nikon a STochastic Optical Reconstruction Microscope (N-STORM) with integrated Fluorescence Lifetime Imaging Microscope (FLIM) system on a 25mm FOV point scanning confocal system. The system provides imaging resolution that overcomes the diffraction-limit (20nm resolution) by using photo-activatable/convertible robust fluorophores. The system will be used to characterize different materials at the single-molecule level, including mapping their catalytic activity. The requested configuration includes a scanning confocal system with attached STochastic Optical Reconstruction Microscopy imaging capabilities, specialized lasers, associated optics, perfect focus and motorized scanners; fully integrated with FLIM for co-localization purposes. The integrated N-STORM, confocal and FLIM microscopies are required to have software able to control all the components of the microscope in wide field, confocal, N-STORM and FLIM modalities simultaneously from a single computer-Software platform and during a single continuous experiment/conditional experiment.
 
PROCESS:
Suppliers that consider their equipment functional, successfully tested, readily available and fully compliant to the ACAN minimum requirements may submit, in writing, a statement of specifications to the contact person identified in this Notice, on or before the closing date of this Notice. In the statement of specifications, the supplier must unequivocally demonstrate how their equipment, at minimum, equals, or exceeds the stated requirements.
 
If no other supplier submits a statement of specifications, on or before the closing date of this Notice, the competitive requirements of the University of Ottawa will be considered having been met. Following notification to any suppliers not successful in unequivocally demonstrating that their statement of specifications equals or exceeds the requirements set out in this Notice, the contract may then be awarded to the pre-identified supplier.
 
Date of issue:       March 5, 2019  
Closing Date:       March 29, 2019, at 3:00:00 P.M. Eastern Standard Time
 
INTENDED USE:
 
The entire setup enables us to perform single-molecule spectroscopy to study materials at the sub-diffraction level, allowing for visualization of molecular interactions at the nanoscopic level. Thus, spatial resolution as high as 20 nm (x, y) can help to determine the position of different active components of a hybrid material whereas the time-resolution imaging allows for tracking their activity in real time. Samples can be mounted into a flow cell specifically designed for fluorescence measurements and the signal from the excited molecules can be collected and sent to an EMCCD camera in order to enable studies of dynamic events in live specimens captured with molecular level resolution. The system includes FLIM and FCS capabilities that can be colocalized with the STORM images. Thus, STORM provides sub-diffraction localization, FCS tells how long a dye stays in the vicinity of the catalytic site and FLIM, by providing fluorescence lifetimes, reveals how the dye interacts with the catalytic site.
 
FUNCTIONALITY:
The equipment proposed by Nikon conforms to the following minimum requirements:
•               The specialized STORM lasers must be CW lasers with outputs at 405, 488, 561 and 647 nm and with the laser power available equal or greater than 100 mW at the exit fiber tip (minimum 125mW for 647nm line). Power stability is an important consideration.
 
•              The optical arrangement should be capable of monitoring broad emission range, including required filter sets.         Thus, 405, 488, 561 and 647 nm filter cubes are necessary. Quad arrangement is an asset as long as it suits        each individual laser/emission wavelength, for instance, working with the 488 nm laser the filter cube must allow detection at all wavelengths above 490 nm.
 
•               Optical arrangement with a broad field of view (FOV): 25 mm without the use of optical adapters
                Repeat position accuracy of at least ±0.5 μm.
 
•               Allow the user to switch seamlessly from widefield to confocal (STORM or FLIM), in experiments designed based on the user’s needs without restarting or changing software application.
 
•               Camera and detector components need to enable fast image acquisition (100 frames/sec). EMCCD camera sensitivity peaking at over 95% Quantum Efficiency, which contributes to allowing STORM imaging of fluorophores with low photon yields. The system must also allow the acquisition of images in up to 6 dimensions (including X, Y, Z, multichannel                 fluorescence/brightfield, large-image stitching, and time lapse within the same experiment and simultaneously). The software platform must possess 3D rendering capabilities as well as the ability to generate movies, animations, and ready-to-go images for presentations.
 
•               Allow for automated, motorized focus control. A piezo z nanopositioner, motorized XY stage with encoders (+/-0.5µm accuracy) and perfect focus system is required to provide unparalleled stability and allow STORM to achieve precise and high 3D spatial resolutions up to ~20 nm (X,Y).
 
•               Two-tiered configuration (stratum structure), modules can be incorporated as two separate layers with multiple modules per layer. Using a dual layer configuration enables optimal filter configuration for each illumination module. This enables optimal filter selection and improves experimental accuracy whilst maintaining the highest acquisition speeds.
 
•               Integrated FLIM with 4 pulsed lasers at 405, 488, 561 and 640 nm and time resolution less than 150 ps.
 
•               FLIM and STORM capabilities must be managed by a single computer by integrated software to allow incorporating both imaging modalities in one acquisition experiment from one software. This will allow us more advanced imaging features in super-resolution. Must include a software environment for designing automated microscopy experiments integrating all modalities of the software.
 
•               FCS (fluorescence correlation spectroscopy) capabilities and appropriate objectives including water immersion objectives.
 
•               4 years of extended warranty for a 5 year total warranty for both STORM and FLIM components.
 
Justification of Pre-Selected Supplier:
Supplier: Nikon
 
Justification:

• Integrated STORM, point scanning confocal, FLIM and FCS: allows for colocalization of species characterized at the sub-diffraction level. STORM provides sub-diffraction localization, FCS tells how long a dye stays in the vicinity of the catalytic site and FLIM, by providing fluorescence lifetimes reveals how the dye interacts with the catalytic site.
• High spatial resolution (in 3D): allows for studies at the single-molecule level.
• Optical arrangement capable of monitoring broad emission range: provides versatility for future expansion of the research topics.
• Large FOV provides more data per experiment, especially for tracking fast processes.
• Fast image acquisition: 100 frames/sec allows time-resolved studies of materials activity and interaction with other molecules, for instance mapping their catalytic activity.