HyU: Hybrid Unmixing for longitudinal in vivo imaging of low signal to noise fluorescence

The expanded application of fluorescence imaging in biomedical and biological research towards more complex systems and geometries requires tools that can analyze a multitude of components at widely varying time- and length-scales. The major challenge in such complex imaging experiments is to cleanly separate multiple fluorescent labels with overlapping spectra from one another and background autofluorescence, without perturbing the sample with high levels of light. Thus, there is a requirement for efficient and robust analysis tools capable of quantitatively separating these signals. In response, we have combined multispectral fluorescence microscopy with hyperspectral phasors and linear unmixing to create Hybrid Unmixing (HyU). Here we demonstrate its capabilities in the dynamic imaging of multiple fluorescent labels in live, developing zebrafish embryos. HyU is more sensitive to low light levels of fluorescence compared to conventional linear unmixing approaches, permitting better multiplexed volumetric imaging over time, with less bleaching. HyU can also simultaneously image both bright exogenous and dim endogenous labels because of its high dynamic range. This allows studies of cellular behaviors, tagged components, and cell metabolism within the same specimen, offering a powerful window into the orchestrated complexity of biological systems.

Digital coded exposure formation of frames from event-based imagery

Event-driven neuromorphic imagers have a number of attractive properties including low-power consumption, high dynamic range, the ability to detect fast events, low memory consumption and low band-width requirements. One of the biggest challenges with using event-driven imagery is that the field of event data processing is still embryonic. In contrast, decades worth of effort have been invested in the analysis of frame-based imagery. Hybrid approaches for applying established frame-based analysis techniques to event-driven imagery have been studied since event-driven imagers came into existence. However, the process for forming frames from event-driven imagery has not been studied in detail. This work presents a principled digital coded exposure approach for forming frames from event-driven imagery that is inspired by the physics exploited in a conventional camera featuring a shutter. The technique described in this work provides a fundamental tool for understanding the temporal information content that contributes to the formation of a frame from event-driven imagery data. Event-driven imagery allows for the application of arbitrary virtual digital shutter functions to form the final frame on a pixel-by-pixel basis. The proposed approach allows for the careful control of the spatio-temporal information that is captured in the frame. Furthermore, unlike a conventional physical camera, event-driven imagery can be formed into any variety of possible frames in post-processing after the data is captured. Furthermore, unlike a conventional physical camera, coded-exposure virtual shutter functions can assume arbitrary values including positive, negative, real, and complex values. The coded exposure approach also enables the ability to perform applications of industrial interest such as digital stroboscopy without any additional hardware. The ability to form frames from event-driven imagery in a principled manner opens up new possibilities in the ability to use conventional frame-based image processing techniques on event-driven imagery.

Simultaneous High Dynamic Range Algorithm, Testing, and Instrument Simulation

Within an imaging instrument’s field of view, there may be many observational targets of interest. Similarly, within a spectrograph’s bandpass, there may be many emission lines of interest. The brightness of these targets and lines can be orders of magnitude different, which poses a challenge to instrument and mission design. A single exposure can saturate the bright emission and/or have a low signal-to-noise ratio (S/N) for faint emission. Traditional high dynamic range (HDR) techniques solve this problem by either combining multiple sequential exposures of varied duration or splitting the light to different sensors. These methods, however, can result in the loss of science capability, reduced observational efficiency, or increased complexity and cost. The simultaneous HDR method described in this paper avoids these issues by utilizing a special type of detector whose rows can be read independently to define zones that are then composited, resulting in areas with short or long exposure measured simultaneously. We demonstrate this technique for the Sun, which is bright on disk and faint off disk. We emulated these conditions in the lab to validate the method. We built an instrument simulator to demonstrate the method for a realistic solar imager and input. We then calculated S/Ns, finding a value of 45 for a faint coronal mass ejection and 200 for a bright one, both at 3.5  ⊙ N —meeting or far exceeding the international standard for digital photography that defines an S/N of 10 as acceptable and 40 as excellent. Future missions should consider this type of hardware and technique in their trade studies for instrument design.

High Dynamic Range Variable Optical Attenuation (Project Report CIAN2-1-Y5)

The goal of this project was to design silicon photonic components for optical WDM (wavelength division multiplexed) add/drop functionality in access and data networks. One of the key functionalities that was required by the campus networking testbed was to control the optical power levels in the WDM network of each channel without requiring separate components. This was most easily achieved by integrating the Variable Optical Attenuator (VOA) functionality together with the add-drop functionality on the same photonic chips.

Innovative Analytical Method for X-ray Imaging and Space-Resolved Spectroscopy of ECR Plasmas

At the Italian National Institute for Nuclear Physics-Southern National Laboratory (INFN-LNS), and in collaboration with the ATOMKI laboratories, an innovative multi-diagnostic system with advanced analytical methods has been designed and implemented. This is based on several detectors and techniques (Optical Emission Spectroscopy, RF systems, interfero-polarimetry, X-ray detectors), and here we focus on high-resolution, spatially resolved X-ray spectroscopy, performed by means of a X-ray pin-hole camera setup operating in the 0.5–20 keV energy domain. The diagnostic system was installed at a 14 GHz Electron Cyclotron Resonance (ECR) ion source (ATOMKI, Debrecen), enabling high-precision, X-ray, spectrally resolved imaging of ECR plasmas heated by hundreds of Watts. The achieved spatial and energy resolutions were 0.5 mm and 300 eV at 8 keV, respectively. Here, we present the innovative analysis algorithm that we properly developed to obtain Single Photon-Counted (SPhC) images providing the local plasma-emitted spectrum in a High-Dynamic-Range (HDR) mode, by distinguishing fluorescence lines of the materials of the plasma chamber (Ti, Ta) from plasma (Ar). This method allows for a quantitative characterization of warm electrons population in the plasma (and its 2D distribution), which are the most important for ionization, and to estimate local plasma density and spectral temperatures. The developed post-processing analysis is also able to remove the readout noise that is often observable at very low exposure times (msec). The setup is now being updated, including fast shutters and trigger systems to allow simultaneous space and time-resolved plasma spectroscopy during transients, stable and turbulent regimes.

Two-Exposure Image Fusion Based on Optimized Adaptive Gamma Correction

In contrast to conventional digital images, high-dynamic-range (HDR) images have a broader range of intensity between the darkest and brightest regions to capture more details in a scene. Such images are produced by fusing images with different exposure values (EVs) for the same scene. Most existing multi-scale exposure fusion (MEF) algorithms assume that the input images are multi-exposed with small EV intervals. However, thanks to emerging spatially multiplexed exposure technology that can capture an image pair of short and long exposure simultaneously, it is essential to deal with two-exposure image fusion. To bring out more well-exposed contents, we generate a more helpful intermediate virtual image for fusion using the proposed Optimized Adaptive Gamma Correction (OAGC) to have better contrast, saturation, and well-exposedness. Fusing the input images with the enhanced virtual image works well even though both inputs are underexposed or overexposed, which other state-of-the-art fusion methods could not handle. The experimental results show that our method performs favorably against other state-of-the-art image fusion methods in generating high-quality fusion results.