Insights into polycrystalline microstructure of blood films with 3D Mueller matrix imaging approach.

This study introduces a novel approach in the realm of liquid biopsies, employing a 3D Mueller-matrix (MM) image reconstruction technique to analyze dehydrated blood smear polycrystalline structures. Our research centers on exploiting the unique optical anisotropy properties of blood proteins, which undergo structural alterations at the quaternary and tertiary levels in the early stages of diseases such as cancer. These alterations manifest as distinct patterns in the polycrystalline microstructure of dried blood droplets, offering a minimally invasive yet highly effective method for early disease detection. We utilized a groundbreaking 3D MM mapping technique, integrated with digital holographic reconstruction, to perform a detailed layer-by-layer analysis of partially depolarizing dry blood smears. This method allows us to extract critical optical anisotropy parameters, enabling the differentiation of blood films from healthy individuals and prostate cancer patients. Our technique uniquely combines polarization-holographic and differential MM methodologies to spatially characterize the 3D polycrystalline structures within blood films. A key advancement in our study is the quantitative evaluation of optical anisotropy maps using statistical moments (first to fourth orders) of linear and circular birefringence and dichroism distributions. This analysis provides a comprehensive characterization of the mean, variance, skewness, and kurtosis of these distributions, crucial for identifying significant differences between healthy and cancerous samples. Our findings demonstrate an exceptional accuracy rate of over 90 % for the early diagnosis and staging of cancer, surpassing existing screening methods. This high level of precision and the non-invasive nature of our technique mark a significant advancement in the field of liquid biopsies. It holds immense potential for revolutionizing cancer diagnosis, early detection, patient stratification, and monitoring, thereby greatly enhancing patient care and treatment outcomes. In conclusion, our study contributes a pioneering technique to the liquid biopsy domain, aligning with the ongoing quest for non-invasive, reliable, and efficient diagnostic methods. It opens new avenues for cancer diagnosis and monitoring, representing a substantial leap forward in personalized medicine and oncology.

Beyond life: Exploring hemodynamic patterns in postmortem mice brains.

We utilize Laser Speckle Contrast Imaging (LSCI) for visualizing cerebral blood flow in mice during and post-cardiac arrest. Analyzing LSCI images, we noted temporal blood flow variations across the brain surface for hours postmortem. Fast Fourier Transform (FFT) analysis depicted blood flow and microcirculation decay post-death. Continuous Wavelet Transform (CWT) identified potential cerebral hemodynamic synchronization patterns. Additionally, non-negative matrix factorization (NMF) with four components segmented LSCI images, revealing structural subcomponent alterations over time. This integrated approach of LSCI, FFT, CWT, and NMF offers a comprehensive tool for studying cerebral blood flow dynamics, metaphorically capturing the 'end of the tunnel' experience. Results showed primary postmortem hemodynamic activity in the olfactory bulbs, followed by blood microflow relocations between somatosensory and visual cortical regions via the superior sagittal sinus. This method opens new avenues for exploring these phenomena, potentially linking neuroscientific insights with mysteries surrounding consciousness and perception at life's end.

Mueller-matrix imaging polarimetry elevated by wavelet decomposition and polarization-singular processing for analysis of specific cancerous tissue pathology.

Significance: Mueller-matrix polarimetry is a powerful method allowing for the visualization of malformations in biological tissues and quantitative evaluation of alterations associated with the progression of various diseases. This approach, in fact, is limited in observation of spatial localization and scale-selective changes in the poly-crystalline compound of tissue samples. Aim: We aimed to improve the Mueller-matrix polarimetry approach by implementing the wavelet decomposition accompanied with the polarization-singular processing for express differential diagnosis of local changes in the poly-crystalline structure of tissue samples with various pathology. Approach: Mueller-matrix maps obtained experimentally in transmitted mode are processed utilizing a combination of a topological singular polarization approach and scale-selective wavelet analysis for quantitative assessment of the adenoma and carcinoma histological sections of the prostate tissues. Results: A relationship between the characteristic values of the Mueller-matrix elements and singular states of linear and circular polarization is established within the framework of the phase anisotropy phenomenological model in terms of linear birefringence. A robust method for expedited (up to ∼15 min) polarimetric-based differential diagnosis of local variations in the poly-crystalline structure of tissue samples containing various pathology abnormalities is introduced. Conclusions: The benign and malignant states of the prostate tissue are identified and assessed quantitatively with a superior accuracy provided by the developed Mueller-matrix polarimetry approach.

Ex vivo confocal Raman microspectroscopy of porcine dura mater supported by optical clearing.

The effect of tissue optical clearing (TOC) to increase the probing depth and observe in-depth structure of the ex vivo porcine dura mater was studied by confocal Raman microspectroscopy (CRM). Raman intensities were significantly increased at the depth of 250 µm for all collagen bands after treatment with glycerol. The influence of glycerol on collagen hydration was also investigated. The results indicate that the process of TOC can be divided into three main steps. The first one is a fast process of tissue dehydration accompanied by collagen shrinkage while the second relatively slow process is related to the glycerol penetration into the interfibrillar space of collagen combined with swelling of tissue. The third step is collagen dissociation caused by the high concentration of glycerol. To the best of our knowledge, this study is the first example to introduce the TOC technique in assisting CRM of ex vivo dura mater in-depth probing.

Time-space Fourier κω' filter for motion artifacts compensation during transcranial fluorescence brain imaging.

Intravital imaging of brain vasculature through the intact cranium in vivo is based on the evolution of the fluorescence intensity and provides an ability to characterize various physiological processes in the natural context of cellular resolution. The involuntary motions of the examined subjects often limit in vivo non-invasive functional optical imaging. Conventional imaging diagnostic modalities encounter serious difficulties in correction of artificial motions, associated with fast high dynamics of the intensity values in the collected image sequences, when a common reference cannot be provided. In the current report, we introduce an alternative solution based on a time-space Fourier transform method so-called K-Omega. We demonstrate that the proposed approach is effective for image stabilization of fast dynamic image sequences and can be used autonomously without supervision and assignation of a reference image.

Combined laser speckle imaging and fluorescent intravital microscopy for monitoring acute vascular permeability reaction.

Optical clearing agents (OCAs) and many chemicals are widely used in functional diagnosis of skin tissues. Numerous studies are associated with the transcutaneous diffusion of OCA in epidermal, dermal, and hypodermal tissues, which results in changing their optical properties. In addition, an objective approach that is suitable for screening the influence of utilized OCA, as well as various chemical agents, synthetics, and nanomaterials, on blood and lymph flows is highly desirable. In our study, a highly sensitive laser speckle imaging (LSI) system and fluorescent intravital microscopy (FIM) were used team-wise to inspect the acute skin vascular permeability reaction in mouse ear during the local application of OCA on the skin surface. Fluorescent contrast material administrated intravenously was used for quantitatively assessing the intensity of vascular permeability reaction and the strength of skin irritation. The obtained results suggest that a combined use of LSI and FIM is highly effective for monitoring the cutaneous vascular permeability reaction, with great potential for assessment of allergic reactions of skin in response to interactions with chemical substances.

Hydrogen bound water profiles in the skin influenced by optical clearing molecular agents-Quantitative analysis using confocal Raman microscopy.

Confocal Raman microscopy has been used to measure depth-dependent profiles of porcine skin ex vivo in the high wavenumber region after application of molecular optical clearing agents (OCAs). Glycerol (70%) and iohexol (100% Omnipaque [300]) water solutions were used as OCAs and topically applied to porcine ear skin for 30 and 60 minutes. Using Gaussian function-based deconvolution, the changes of hydrogen bound water molecule types have been microscopically analyzed down to the depth of 200 µm. Results show that both OCAs induced skin dehydration (reduction of total water), which is 51.3% for glycerol (60 minutes), 33.1% for glycerol (30 minutes), 8.3% for Omnipaque (60 minutes) and 4.4% for Omnipaque (30 minutes), on average for the 40 to 200 µm depths. Among the water types in the skin, the following reduction was observed in concentration of weakly bound (51.1%, 33.2%, 7.5% and 4.6%), strongly bound (50.4%, 33.0%, 7.9% and 3.4%), tightly bound (63.6%, 42.3%, 26.1% and 12.9%) and unbound (55.4%, 28.7%, 10.1% and 5.9%) water types on average for the 40 to 200 µm depths, post application of glycerol (60 minutes), glycerol (30 minutes), Omnipaque (60 minutes) and Omnipaque (30 minutes), respectively. As most concentrated in the skin, weakly and strongly bound water types are preferentially involved in the OCA-induced water flux in the skin, and thus, are responsible for optical clearing efficiency.

A comparative study of ex vivo skin optical clearing using two-photon microscopy.

Multiphoton tomography (MPT) is a prospective tool for imaging the skin structure. Aiming to increase the probing depth, a comparative ex vivo study of optical clearing of porcine ear skin was performed by using two optical clearing agents (OCAs), i.e., glycerol and iohexol (OmnipaqueTM ) at different concentrations, which exhibit different osmotic properties. The results show that a topical application of glycerol or OmnipaqueTM solutions onto the skin for 60 min significantly improved the depth and contrast of the MPT signals. By utilizing 40%, 60% and 100% glycerol, and 60% and 100% OmnipaqueTM it was demonstrated that both agents improve autofluorescence and SHG (second harmonic generation) signals from the skin. At the applied concentrations and agent time exposure, glycerol is more effective than OmnipaqueTM . However, tissue shrinkage and cell morphology changes were found for highly concentrated glycerol solutions. OmnipaqueTM , on the contrary, increases the safety and has no or minimal tissue shrinkage during the optical clearing process. Moreover OmnipaqueTM allows for robust multimodal optical/X-ray imaging with automatically matched optically cleared and X-ray contrasted tissue volumes. These findings make OmnipaqueTM more prospective than glycerol for some particular application.