Combined approach using circular intensity differential scattering microscopy under phasor map data analysis

Abstract

Circular intensity differential scattering (CIDS) is based on the analysis of circular polarized light scattering and has been proven to be an interesting label-free microscopy technique sensitive to the chiral organization at the submicroscopic level. However, this approach averages the localized contrasts related to the sample polarimetric properties in the illumination volume. Additionally, the detection sensitivity suffers from the confinement of the mixture of structures, and it becomes an arduous task to discriminate the source of the signal. In this work, we show that a phasor map approach combined with CIDS microscopy has provided an intuitive view of the sample organization to recognize the presence of different molecular species in the illumination volume. The data represented in terms of polarization response mapped to a single point called a phasor also have the potential to pave the way for the analysis of large data sets. We validated this method by numerical simulations and compared the results with that of experimental data of optical devices of reference.

Roadmap on Label‐Free Super‐Resolution Imaging

“…[ 371 ] The phasor representation can be used to allow medical doctors and clinicians to visualize the possible presence of a classified population in a friendly and effective way. [ 380 ] Figure 20c shows an example based on a population classification. Figure 20d shows the label‐free ability of detecting a different compaction of chromatin in the cell nucleus using “classical” fluorescence DNA signature as benchmark.…”

“…[371] The phasor representation can be used to allow medical doctors and clinicians to visualize [374] b) An example of theoretical results about the presence of a spike-virus in a spherical droplet with size comparable to sars-cov2. [378] c) Use of the Phasor analysis to distinguish species on the basis of CIDS signature [380] ; d) Normalized CIDS (blu) image of an isolated HEK nucleus after extraction compared with fluorescence intensity (green) signature based on DNA labeling. [374] Labels: Ti:Sa: Titanium-Sapphire coherent laser source tuned at 740 nm to prime both CIDS and two-photon excitation fluorescence.…”

Phasor map analysis to investigate Hutchinson–Gilford progeria cell under polarization-resolved optical scanning microscopy

“…In our previous work, we have proposed for the first time a simple theoretical study based on a phasor data analysis that exhibits remarkable capability for recognizing different structures that exhibit a strong birefringent and dichroic response in turbid medium. In parallel, we have propose to apply this approach by measuring the polarimetric fingerprint of isolated cell nuclei in a single point spectroscopic architecture by acquiring the angular scattering modulated signal using a rotational detection arm 20 . Here, we proposed to adapt our previous approach into an imaging architecture and we show that it provides on a robust and distinctive analytical approach sensitive to the polarimetric variations of different type of particle located into the PSF volume.…”

Phasor approach of Mueller matrix optical scanning microscopy for biological tissue imaging

“…In the context of nonlinear, label-free microscopy, the phasor plot has been used to analyze polarization-resolved second harmonic generation data and map the features of collagen architecture in tissues ( 27 ). Moreover, a recent theoretical work has proven the phasor space based on the analysis of circular polarized light scattering to be an intuitive tool, sensitive to the chiral organization of biopolymers at the submicroscopic level ( 28 ).…”