skip to main content
US FlagAn official website of the United States government
dot gov icon
Official websites use .gov
A .gov website belongs to an official government organization in the United States.
https lock icon
Secure .gov websites use HTTPS
A lock ( lock ) or https:// means you've safely connected to the .gov website. Share sensitive information only on official, secure websites.


Title: Dual-angle light scattering spectroscopy for calibration-free measurements of scatterer suspensions
It can be difficult to employ optical techniques for analyzing biological structures smaller than or comparable to the wavelength of light, such as extracellular vesicles or some types of bacteria. Biological light scattering spectroscopy (LSS), developed to address this problem, has been successfully used for characterizing tissue on cellular and subcellular scales. At the same time, calibration with a reference sample of known optical properties can complicate LSS measurements. In this Letter, we present dual-angle LSS (daLSS), which is designed for calibration-free measurements of scatterer suspensions. It employs measurements of a sample at two distinct angles, which then allows system effects to be removed entirely. Not only does daLSS simplify and speed up the measurement procedure, but it also makes spectra more reproducible, an important feature for diagnostic techniques. We validated the technique by accurately reconstructing the sizes of polystyrene microspheres with diameters less than 100 nm and then demonstrated that not only are the daLSS spectra of several common bacteria strains easily distinguishable but they are also highly reproducible.  more » « less
Award ID(s):
1948722 2220273 2325317
PAR ID:
10560408
Author(s) / Creator(s):
; ; ; ; ; ; ; ;
Publisher / Repository:
Optical Society of America
Date Published:
Journal Name:
Optics Letters
Volume:
50
Issue:
1
ISSN:
0146-9592; OPLEDP
Format(s):
Medium: X Size: Article No. 33
Size(s):
Article No. 33
Sponsoring Org:
National Science Foundation
More Like this
  1. Extremely variable quasars can also show strong changes in broad-line emission strength and are known as changing-look quasars (CLQs). To study the CLQ transition mechanism, we present a pilot sample of CLQs with X-ray observations in both the bright and faint states. From a sample of quasars with bright-state archival SDSS spectra and (Chandra or XMM-Newton) X-ray data, we identified five new CLQs via optical spectroscopic follow-up and then obtained new target-of-opportunity X-ray observations with Chandra. No strong absorption is detected in either the bright- or the faint-state X-ray spectra. The intrinsic X-ray flux generally changes along with the optical variability, and the X-ray power-law slope becomes harder in the faint state. Large-amplitude mid-infrared variability is detected in all five CLQs, and it echoes the variability in the optical with a time lag expected from the light-crossing time of the dusty torus for CLQs with robust lag measurements. The changing-obscuration model is not consistent with the observed X-ray spectra and spectral energy distribution changes seen in these CLQs. It is highly likely that the observed changes are due to the changing accretion rate of the supermassive black hole, so the multiwavelength emission varies accordingly, with promising analogies to the accretion states of X-ray binaries. 
    more » « less
  2. Programmable illumination control is essential for many computational microscopy techniques. Conventional light source array is often arranged on a fixed grid of a planar surface for providing programmable sample illumination. Here, we report the development of a freeform illuminator that can be arranged at arbitrary 2-dimensional or 3-dimensional (3D) surface structures for computational microscopy. The freeform illuminator can be designed in a small form factor with a dense light source arrangement in 3D. It can be placed closer to the sample for providing angle-varied illumination with higher optical flux and smaller angular increment. With the freeform illuminators, we develop a calibration process using a low-cost Raspberry-Pi image sensor coated with a monolayer of blood cells. By tracking the positional shift of the blood-cell diffraction patterns at 2 distinct regions of the coded sensor, we can infer the 3D positions of the light source elements in a way similar to the stereo vision reconstruction approach. To demonstrate the applications for computational microscopy, we validate the freeform illuminators for Fourier ptychographic microscopy, 3D tomographic imaging, and on-chip microscopy. We also present a longitudinal study by tracking the growth of live bacterial cultures over a large field of view. The reported freeform illuminators and the related calibration process offer flexibilities and extended scope for imaging innovations in computational microscopy. 
    more » « less
  3. Random Forests (RFs) are at the cutting edge of supervised machine learning in terms of prediction performance, especially in genomics. Iterative RFs (iRFs) use a tree ensemble from iteratively modified RFs to obtain predictive and stable nonlinear or Boolean interactions of features. They have shown great promise for Boolean biological interaction discovery that is central to advancing functional genomics and precision medicine. However, theoretical studies into how tree-based methods discover Boolean feature interactions are missing. Inspired by the thresholding behavior in many biological processes, we first introduce a discontinuous nonlinear regression model, called the “Locally Spiky Sparse” (LSS) model. Specifically, the LSS model assumes that the regression function is a linear combination of piecewise constant Boolean interaction terms. Given an RF tree ensemble, we define a quantity called “Depth-Weighted Prevalence” (DWP) for a set of signed features S ± . Intuitively speaking, DWP( S ± ) measures how frequently features in S ± appear together in an RF tree ensemble. We prove that, with high probability, DWP( S ± ) attains a universal upper bound that does not involve any model coefficients, if and only if S ± corresponds to a union of Boolean interactions under the LSS model. Consequentially, we show that a theoretically tractable version of the iRF procedure, called LSSFind, yields consistent interaction discovery under the LSS model as the sample size goes to infinity. Finally, simulation results show that LSSFind recovers the interactions under the LSS model, even when some assumptions are violated. 
    more » « less
  4. Features such as particles, pores, or cracks are challenging to measure accurately in CT data when they are small relative to the data resolution, characterized as a point-spread function (PSF). These challenges are particularly acute when paired with segmentation, as the PSF distributes some of the signal from a voxel among neighboring ones; effectively dispersing some of the signal from a given object to a region outside of it. Any feature of interest with one or more dimensions on the order of the PSF will be impacted by this effect, and measurements based on global thresholds necessarily fail. Measurements of the same features should be consistent across different instruments and data resolutions. The PVB (partial volume and blurring) method successfully compensates by quantifying features that are small in all three dimensions based on their attenuation anomaly. By calibrating the CT number of the phase of interest (in this case, gold) it is possible to accurately measure particles down to <6 voxels in data acquired on two instruments, 14 years apart, despite severe artifacts. Altogether, the PVB method is accurate, reproducible, resolution-invariant, and objective; it is also notable for its favorable error structure. The principal challenge is the need for representative effective CT numbers, which reflect not only the features of interest themselves, but also the X-ray spectrum, the size, shape and composition of the enclosing sample, and processing details such as beam-hardening correction. Empirical calibration is the most effective approach. 
    more » « less
  5. North-Morris, Michael B.; Creath, Katherine; Porras-Aguilar, Rosario (Ed.)
    A novel Vision ray metrology technique is reported that estimates the geometric wavefront of a measurement sample using the sample-induced deflection in the vision rays. Vision ray techniques are known in the vision community to provide image formation models even when conventional camera calibration techniques fail. This work extends the use of vision rays to the area of optical metrology. In contrast to phase measuring deflectometry, this work relies on differential measurements, and hence, the absolute position and orientation between target and camera do not need to be known. This optical configuration significantly reduces the complexity of the reconstruction algorithms. The proposed vision ray metrology system does not require mathematical optimization algorithms for calibration and reconstruction – the vision rays are obtained using a simple 3D fitting of a line. 
    more » « less