In time-lapse optical microscopy of multiple field of views (FOVs), there is a trade-off between the spatial coverage that can be achieved at high spatial resolution and the temporal resolution. In other words, the larger the spatial coverage, the longer it takes to acquire all small (FOVs) to create one large FOV. Consequently, the events in a specimen that are characterized by high rates of changes that require high temporal resolution cannot be captured over a large FOV (e.g.., at population levels).
To address this tradeoff, NIST is interested in designs (Phase I) and prototypes (Phase II) of an optical microscope that would enable large FOV imaging of dynamic events and meet the specifications provided by the project goals.
Included are a few references [1-4] that describe some of the published designs addressing the current tradeoff. Proposers are encouraged to innovate published designs or pursue their own new designs.
The project goals are to enable microscopy measurements that could assist in characterizing dynamic events over entire populations of mammalian cells. One such example would be time-lapse imaging of live cell cultures.
To enable such measurements, the microscope design and its prototype should aim at meeting the following specifications:
Resolution: 1.4 μm or smaller when using the imaging system in a non-automated fashion and manually focusing on an object. This figure was arrived at by computing the Abbe diffraction limited resolution given by LAbbe = λ/2NA, where λ=550 nm, NA = 0.3 and allowing for a 50% deviation from ideality. The measurement of resolution can be performed using the method described by Vainrub5 or an equivalent method. Preference will be given to solutions that provide for similar resolutions with functioning in an automated mode with auto-focusing.
Spatial coverage and acquisition time: Solutions should be capable of imaging a 1 cm2 area/60 seconds.
Modality: Widefield or confocal solutions (solutions that include also transmitted light imaging are preferred).
Modularity: Plug-and-play component to an automated microscope equipped with a scanning stage and focus control.
Material cost: < $20K
Phase I expected results:
Design an optical microscope that could acquire high-resolution large-FOV images “near real time” as specified by the project goals.
Phase II expected results:
Prototype the designed optical microscope
NIST may be available to discuss microscope design approaches and to provide additional specific inputs about a variety of demanding imaging applications at NIST.