ABSTRACT
This work compares two tomographic imaging technologies, time-domain full-field optical coherence tomography (FFOCT) working in reflection and optical transmission tomography (OTT), using a new optical setup that combines both. We show that, due to forward-scattering properties, the axial sectioning and contrast in OTT can be optimized by tuning illumination. The influence of sample scattering and thickness are discussed. We illustrate the comparison of the two methods in static (morphology) and dynamic (metabolic contrast) regimes using cell cultures, tissues and entire organisms emphasizing the advantages of both approaches.
ABSTRACT
Viruses have a profound influence on all forms of life, motivating the development of rapid and minimally invasive methods for virus detection. In this study, we present a novel methodology that enables quantitative measurement of the interaction between individual biotic nanoparticles and antibodies in solution. Our approach employs a label-free, full-field common-path interferometric technique to detect and track biotic nanoparticles and their interactions with antibodies. It is based on the interferometric detection of light scattered by viruses in aqueous samples for the detection of individual viruses. We employ single-particle tracking analysis to characterize the size and properties of the detected nanoparticles, and to monitor the changes in their diffusive mobility resulting from interactions. To validate the sensitivity of our detection approach, we distinguish between particles having identical diffusion coefficients but different scattering signals, using DNA-loaded and DNA-devoid capsids of the Escherichia coli T5 virus phage. In addition, we have been able to monitor, in real time, the interaction between the bacteriophage T5 and purified antibodies targeting its major capsid protein pb8, as well as between the phage SPP1 and nonpurified anti-SPP1 antibodies present in rabbit serum. Interestingly, these virus-antibody interactions are observed within minutes. Finally, by estimating the number of viral particles interacting with antibodies at different concentrations, we successfully quantify the dissociation constant Kd of the virus-antibody reaction using single-particle tracking analysis.
ABSTRACT
There is an increasing need for label free methods that could reveal intracellular structures and dynamics. In this context, we develop a new optical tomography method working in transmission - full-field optical transmission tomography (FF-OTT). The method can measure the forward scattering signals and reveals the time-dependent metabolic signals in living cells. FF-OTT is a common path interferometer taking advantage of the Gouy phase shift - a π phase shift that the light wave experiences around the focus. By modulating the position of the focus one can alter the phase of the scattered light. Demodulation of images with different phases rejects the background and enhances the light from the depth-of-field, thus producing an optical section. We test FF-OTT by imaging single-cell diatoms and ex vivo biological samples. In fresh samples, we show that the intracellular motions create visible intensity fluctuations in FF-OTT so that the method is able to reveal a metabolic dynamic contrast. FF-OTT was found to be an efficient label free technique that can be readily implemented thanks to a robust common-path speckle-free interferometer design using an incoherent light source.
ABSTRACT
We study a double-scattering coherent mechanism of negative polarization (NP) near opposition that is observed for powder-like surfaces. The problem is solved numerically for absorbing structures with irregular constituents, cubes, spheres, and ellipsoids larger than the wavelength of incident light. Our simulations show that double scattering between two random irregular particles shows weak NP. Adding one more particle significantly increases the relative contribution of double scattering which enhances NP. Simulations with regular shapes and controlled geometric parameters show that the interference mechanism is sensitive to the geometry of the scattering system and can also result in no polarization or even strong enhancement of positive polarization at backscattering.
