Laser tissue welding by using collagen excitation at a 1,720 nm near-infrared optical window III.

Laser tissue welding (LTW) is a method of fusing incised tissues together. LTW has the potential to revolutionize plastic surgery and wound healing techniques by its ability to produce watertight, scarless seals with minimal foreign body reaction. While using thermal mechanisms to achieve LTW, energy from the incident laser is absorbed by water in the tissue. As the water temperature increases, partial denaturing of the collagen triple helix briefly occurs, which is quickly followed by renaturation of collagen as the tissue cools, thus providing a watertight seal. This research study investigates the efficacy of direct collagen excitation at 1,720 nm to accomplish LTW. This wavelength falls within the near-infrared (NIR) optical window III. The tensile strengths of pig skin that have been welded with NIR continuous-wave (CW) diode lasers at 1,455 nm, which promote thermal mechanisms of tissue welding, and 1,720 nm wavelengths, are compared. Near-infrared lasers tuned to 1,455 and 1,720 nm were used to weld incised pieces of porcine skin together without extrinsic solders or dyes. The tensile force of the welded tissues was measured using a digital force gauge. The average tensile force of the welded pig skin using the 1,720 nm laser was approximately four times greater than that using the CW 1,455 nm laser, suggesting that LTW accomplished through direct collagen excitation in the NIR optical window III provides greater tensile strengths.

Femtosecond optical Kerr effect in normal and grades of cancerous breast tissues as a new optical biopsy method.

This study reports on the first use of the optical Kerr effect (OKE) in breast cancer tissue. This proposed optical biopsy method utilizes a Femtosecond Optical Kerr Gate to detect changes in dielectric relaxation and conductivity created by a cancerous infection. Here, the temporal behavior of the OKE is tracked in normal and cancerous samples of human and mouse breast. These tissues display a double peaked temporal structure and its decay rate changes depending on the tissue's infection status. The decay of the secondary peak, attributed to ultrafast plasma response, indicates that the tissue's conductivity has doubled once infected. A slower molecular contribution to the Kerr effect can also be observed in healthy tissues. These findings suggest two possible biomarkers for the use of OKE in optical biopsy. Both markers arise from alterations in the infected tissue's cellular structure, which changes the rate at which electronic and molecular processes occur.

Higher harmonics and supercontinuum generated from the Kerr response time in different states of matter from a universal electromagnetic model.

There is a need for a universal model to describe higher harmonic generation (HHG) in different states of matter. Based on an electromagnetic model (EM), the generation of odd higher harmonic (HHG) and supercontinuum (SC) from intense fs and ps pulses for visible, NIR, and MIR lasers is simulated based on the parameters from experimental observation. HHG and SC depend critically on the different Kerr material response times τ from the ultrafast on the order of 100 as for electronic cloud distortion to fast ~ 10 fs from plasma and molecular redistribution and to the slower picoseconds rotational and vibrational molecular processes. The number of odd HHG generated is shown to depend critically on the fastest Kerr response time on the order of ~ 1 fs from electronic self-phase modulation (ESPM). In this study, different states of matter from noble gas Argon to condensed matter ZnO and LBG are simulated showing the dependence on the Kerr response time to produce HHG for various applications in Physics, Chemistry, Biology, and Engineering. The EM model is universal to produce HHG and SC in different states of matter.

Femtosecond Optical Kerr Gate in tissues.

The Optical Kerr Effect is investigated for the first time in biological tissues. This nonlinear effect was explored in both human brain and avian breast tissues using a time-resolved femtosecond pump-probe Optical Kerr Gate that looks for phase changes that arise in the probe from the pump induced Kerr refractive index change. The tissue samples produced a unique ultrafast (700-800 fs) doubled peaked temporal signal, which is indicative of interplay between the different ultrafast mechanisms (electronic plasma and molecular) that make up the Kerr index. The unique profile was replicated in theoretical simulations. The properties of the temporal profile varied between samples suggesting that it could be used as a new diagnostic. Understanding this behavior can help improve the scientific understanding of nonlinear spectral diagnostic techniques and potentially create a new Kerr-based optical biopsy method.

Femtosecond Optical Kerr Gates in Cancerous Breast Tissue for a New Optical Biopsy Method.

The Optical Kerr Effect was demonstrated for the first time as a new optical biopsy method to detect normal and grades of cancer of human breast tissues. The technique works by temporally tracking the various electronic and molecular processes that give rise to the nonlinear index of refraction (n2). The rate at which these processes populate and dissipate varies depending on the internal properties of the sample. It is shown here that in tissues, the variances in the ultrafast plasma Kerr responses that relates to the dielectric relaxation can be used as a biomarker for cancer. The relaxation of this response changes significantly between healthy and different grades of triple negative breast cancer tissues. This change can be attributed to a doubling or tripling of the tissue's conductivity depending on the cancer grade.

A Handheld Visible Resonance Raman Analyzer Used in Intraoperative Detection of Human Glioma.

There is still a lack of reliable intraoperative tools for glioma diagnosis and to guide the maximal safe resection of glioma. We report continuing work on the optical biopsy method to detect glioma grades and assess glioma boundaries intraoperatively using the VRR-LRRTM Raman analyzer, which is based on the visible resonance Raman spectroscopy (VRR) technique. A total of 2220 VRR spectra were collected during surgeries from 63 unprocessed fresh glioma tissues using the VRR-LRRTM Raman analyzer. After the VRR spectral analysis, we found differences in the native molecules in the fingerprint region and in the high-wavenumber region, and differences between normal (control) and different grades of glioma tissues. A principal component analysis-support vector machine (PCA-SVM) machine learning method was used to distinguish glioma tissues from normal tissues and different glioma grades. The accuracy in identifying glioma from normal tissue was over 80%, compared with the gold standard of histopathology reports of glioma. The VRR-LRRTM Raman analyzer may be a new label-free, real-time optical molecular pathology tool aiding in the intraoperative detection of glioma and identification of tumor boundaries, thus helping to guide maximal safe glioma removal and adjacent healthy tissue preservation.

Stokes shift spectroscopy and machine learning for label-free human prostate cancer detection.

The Stokes shift spectra (S3) of human cancerous and normal prostate tissues were collected label free at a selected wavelength interval of 40 nm to investigate the efficacy of the approach based on three key molecules-tryptophan, collagen, and reduced nicotinamide adenine dinucleotide (NADH)-as cancer biomarkers. S3 combines both fluorescence and absorption spectra in one scan. The S3 spectra were analyzed using machine learning (ML) algorithms, including principal component analysis (PCA), nonnegative matrix factorization (NMF), and support vector machines (SVMs). The components retrieved from the S3 spectra were considered principal biomarkers. The differences in the weights of the components between the two types of tissues were found to be significant. Sensitivity, specificity, and accuracy were calculated to evaluate the performance of SVM classification. This research demonstrates that S3 spectroscopy is effective for detecting the changes in the relative concentrations of the endogenous fluorophores in tissues due to the development of cancer label free.

Fano resonance line shapes in the Raman spectra of tubulin and microtubules reveal quantum effects.

Microtubules are self-assembling biological nanotubes made of the protein tubulin that are essential for cell motility, cell architecture, cell division, and intracellular trafficking. They demonstrate unique mechanical properties of high resilience and stiffness due to their quasi-crystalline helical structure. It has been theorized that this hollow molecular nanostructure may function like a quantum wire where optical transitions can take place, and photoinduced changes in microtubule architecture may be mediated via changes in disulfide or peptide bonds or stimulated by photoexcitation of tryptophan, tyrosine, or phenylalanine groups, resulting in subtle protein structural changes owing to alterations in aromatic flexibility. Here, we measured the Raman spectra of a microtubule and its constituent protein tubulin both in dry powdered form and in aqueous solution to determine if molecular bond vibrations show potential Fano resonances, which are indicative of quantum coupling between discrete phonon vibrational states and continuous excitonic many-body spectra. The key findings of this work are that we observed the Raman spectra of tubulin and microtubules and found line shapes characteristic of Fano resonances attributed to aromatic amino acids and disulfide bonds.

Intraoperative detection of human meningioma using a handheld visible resonance Raman analyzer.

To report for the first time the preliminary results for the evaluation of a VRR-LRR™ analyzer based on visible resonance Raman technique to identify human meningioma grades and margins intraoperatively. Unprocessed primary and recurrent solid human meningeal tissues were collected from 33 patients and underwent Raman analysis during surgeries. A total of 1180 VRR spectra were acquired from fresh solid tissues using a VRR-LRR™ analyzer. A confocal HR Evolution (HORIBA, France SAS) Raman system with 532-nm excitation wavelength was also used to collect data for part of the ex vivo samples after they were thawed from - 80 °C for comparison. The preliminary analysis led to the following observations. (1) The intensity ratio of VRR peaks of protein to fatty acid (I2934/I2888) decreased with the increase of meningioma grade. (2) The ratio of VRR peaks of phosphorylated protein to amid I (I1588/I1639) decreased for the higher grade of meningioma. (3) Three RR vibration modes at 1378, 3174, and 3224 cm-1 which were related to the molecular vibrational bands of oxy-hemeprotein, amide B, and amide A protein significantly changed in peak intensities in the two types of meningioma tissues compared to normal tissue. (4) The changes in the intensities of VRR modes of carotenoids at 1156 and 1524 cm-1 were also found in the meningioma boundary. The VRR-LRR™ analyzer demonstrates a new approach for label-free, rapid, and objective identification of primary human meningioma in quasi-clinical settings. The accuracy for detecting meningioma tissues using support vector machines (SVMs) was over 70% based on Raman peaks of key biomolecules and up to 100% using principal component analysis (PCA).

Photosynthesis on the Ultrafast Time Scale within the Confinements of Quantum Nanostructured Photosystems.

Fundamental information on the behavior of excited chlorophyll molecules packed within the confinements of nanosized photosystems I and II, following absorption of light, is presented. Using a 100 femtosecond laser with nanojoule (nJ) pulse energy and a one picosecond streak camera, we observed the light emitted from the nanostructured photosystems without oscillations or hops. The fluorescent exponential decay profiles and high efficiency within the nanostructure suggest that light coherently drains out as a unit. This implies that "quantumness" is linked to quantum confinement on the nano scale.

Femtosecond anti-Stokes conical emission in BK-7 glass and O and E-wave calcite.

The angle of anti-Stokes conical emission (CE) is experimentally measured in the frequency shift span of 2000cm-1 to 9000cm-1. The experiment was performed using a 800 nm 50 fs laser pump in samples of BK-7 glass and calcite in both the O and E-wave configurations. The experimental results of angular emission are then compared to three competing models: the Alfano-Shapiro four wave mixing (FWM) model from 1970, the Luther FWM model from 1994, and the Faccio X-wave model from 2004. Results indicate that in all samples and configurations tested, the original FWM has the best agreement with experimental results in the anti-Stokes span.

Enhanced stimulated raman scattering of solvent due to anharmonic energy transfer from resonance raman solute molecules.

A new nonlinear optical process, named enhanced stimulated Raman scattering (ESRS), is reported for the first time from resonance Raman in ß-carotene-methanol solution. It is well known that absorption decreases the efficiency of the nonlinear optical and laser processes; however, we observed enhanced stimulated Raman peaks at the first and second Stokes from methanol solvent at 2834 cm-1 with the addition of ß-carotene solutes. This enhanced SRS effect in methanol is attributed to the resonance Raman (RR) process in ß-carotene, which creates a significant number of vibrations from RR and the excess vibrations are transferred to methanol from anharmonic vibrational interactions between the ß-carotene solutes and the methanol solvent, and consequently leads to the increased Raman gain.

Steady-state stimulated Raman generation and filamentation using complex vector vortex beams.

Stimulated Raman scattering and laser filamentation produced using nanosecond pulsed complex vector vortex beams (CVVB) are investigated in a 20 cm long methanol cell. The CVVB is generated using q-plates and is tested at orbital angular momentum (L) values of 1, 2, 3, and 4 and circular, radial, and azimuthal polarizations. The results illustrate that the stability and intensity of the generated stimulated Raman has dependence on input polarization and L value. During filamentation, the beam is also shown to break up into multiple primary filaments and that there is a reduction in small-scale filamentation when using CVVBs.

Modulation instability induced by cross-phase modulation of transient stimulated Raman scattering and self-phase modulation in calcite.

Pairs of sidebands about the transient stimulated Raman scattering (SRS) 1086cm-1 vibration mode peak are observed for calcite under 517 nm 390 fs pulse excitation. These pairs of side frequency lobes arise from modulation instability (MI) from the interaction of cross-phase modulation (XPM) from self-phase modulation (SPM) and SRS. The pairs of secondary frequencies are attributed to the daughter 1086cm-1 decay product modes from the multiphonon of 3, 4, and 5 decays. The main sideband peak from 1086cm-1 phonon at 546cm-1 suggests the operation of the Orbach effect.

Explanation of the competition between O- and E-wave induced stimulated Raman and supercontinuum in calcite under ultrafast laser excitation.

Key optical properties of calcite were measured to unravel the difference between stimulated Raman scattering (SRS) and self-phase modulation (SPM) for the supercontinuum (SC) for ordinary (O) wave and extraordinary (E) wave. These properties are group velocity dispersion, walk-off, spontaneous Raman spectra and cross section, optical 1086cm-1 phonon linewidth, nonlinear susceptibility (χ3), steady-state and transient SRS, and SC caused from SPM. These are investigated for O-waves and E-waves from a 2.7 cm thick calcite crystal. Using 390 fs pulses (∼0.8µJ pulse energy) at 517 nm, the O-wave produced a stronger sharp SRS peak at 1086cm-1 and a weaker SC spectrum in the visible range than the E-wave. The salient difference found between the O- and E-waves for SRS and SPM in calcite is attributed to the larger Raman cross section and the size of nonlinear susceptibility (χ3) for O-waves as compared to E-waves.

Comparison of continuous wave versus picosecond SRS and the resonance SRS effect.

Stimulated Raman scattering (SRS) is a powerful optical technique for probing the vibrational states of molecules in biological tissues and provides greater signal intensities than when using spontaneous Raman scattering. In this study, we examined the use of continuous wave (cw) and picosecond (ps) laser excitations to generate SRS signals in pure methanol, a carotene-methanol solution, acetone, and brain tissue samples. The cw-SRS system, which utilized two cw lasers, produced better signal-to-noise (S/N) than the conventional ps-SRS system, suggesting that the cw-SRS system is an efficient and cost-effective approach for studying SRS in complex systems like the brain. The cw-SRS approach will reduce the size of the SRS system, allowing for stimulated Raman gain/loss microscopy. In addition, we showed that there exists a resonance SRS (RSRS) effect from the carotene-methanol solution and brain tissue samples using cw laser excitations. The RSRS effect will further improve the signal-to-noise and may be utilized as an enhanced, label-free SRS microscopic tool for the study of biological tissues.

Distinguishing metastatic triple-negative breast cancer from nonmetastatic breast cancer using second harmonic generation imaging and resonance Raman spectroscopy.

Triple-negative breast cancer (TNBC) is an aggressive subset of breast cancer that is more common in African-American and Hispanic women. Early detection followed by intensive treatment is critical to improving poor survival rates. The current standard to diagnose TNBC from histopathology of biopsy samples is invasive and time-consuming. Imaging methods such as mammography and magnetic resonance (MR) imaging, while covering the entire breast, lack the spatial resolution and specificity to capture the molecular features that identify TNBC. Two nonlinear optical modalities of second harmonic generation (SHG) imaging of collagen, and resonance Raman spectroscopy (RRS) potentially offer novel rapid, label-free detection of molecular and morphological features that characterize cancerous breast tissue at subcellular resolution. In this study, we first applied MR methods to measure the whole-tumor characteristics of metastatic TNBC (4T1) and nonmetastatic estrogen receptor positive breast cancer (67NR) models, including tumor lactate concentration and vascularity. Subsequently, we employed for the first time in vivo SHG imaging of collagen and ex vivo RRS of biomolecules to detect different microenvironmental features of these two tumor models. We achieved high sensitivity and accuracy for discrimination between these two cancer types by quantitative morphometric analysis and nonnegative matrix factorization along with support vector machine. Our study proposes a new method to combine SHG and RRS together as a promising novel photonic and optical method for early detection of TNBC.

Optical biopsy identification and grading of gliomas using label-free visible resonance Raman spectroscopy.

Glioma is one of the most refractory types of brain tumor. Accurate tumor boundary identification and complete resection of the tumor are essential for glioma removal during brain surgery. We present a method based on visible resonance Raman (VRR) spectroscopy to identify glioma margins and grades. A set of diagnostic spectral biomarkers features are presented based on tissue composition changes revealed by VRR. The Raman spectra include molecular vibrational fingerprints of carotenoids, tryptophan, amide I/II/III, proteins, and lipids. These basic in situ spectral biomarkers are used to identify the tissue from the interface between brain cancer and normal tissue and to evaluate glioma grades. The VRR spectra are also analyzed using principal component analysis for dimension reduction and feature detection and support vector machine for classification. The cross-validated sensitivity, specificity, and accuracy are found to be 100%, 96.3%, and 99.6% to distinguish glioma tissues from normal brain tissues, respectively. The area under the receiver operating characteristic curve for the classification is about 1.0. The accuracies to distinguish normal, low grade (grades I and II), and high grade (grades III and IV) gliomas are found to be 96.3%, 53.7%, and 84.1% for the three groups, respectively, along with a total accuracy of 75.1%. A set of criteria for differentiating normal human brain tissues from normal control tissues is proposed and used to identify brain cancer margins, yielding a diagnostic sensitivity of 100% and specificity of 71%. Our study demonstrates the potential of VRR as a label-free optical molecular histopathology method used for in situ boundary line judgment for brain surgery in the margins.

Majorana vortex photons a form of entangled photons propagation through brain tissue.

This paper extends the concept of entangled vector vortex beams as a form of Majorana-like photons. Majorana photon quasi particles are introduced and attributed to a class of entangled vector beams and show higher transmission. These photons and the antiphotons are identical. A Majorana photon has within itself both right and left handed twists. These majorana beams travel at speeds other than speed of light, c in free space. Light transmission of Majorana photon vortex beams with orbital angular momentum (OAM) are investigated in a mouse brain at different local regions showing enhanced transmission and properties of being entangled. This work is new interpretation of our past paper of mixed photon beam states. The transmission change observed with Majorana structured light other than linear polarization is attributed to the nonseparable and mixed nature of radial and azimuthal polarizations with OAM and the handedness of the light passing through chiral brain media. These mixed nonhomogeneous beams are entangled in OAM and polarization. Majorana photons may play an important role in the future for quantum and optical computing and sub and super luminal speeds due to its traversal wave vector, k.

Transmission of classically entangled beams through mouse brain tissue.

Light transmission of Laguerre-Gaussian vector vortex beams in different local regions in mouse brain tissue is investigated. Transmittance is measured in the ballistic and diffusive regions with various polarizations states and orbital angular momentums (OAM). The transmission change observed with structured light other than linear polarization is attributed to chiroptical phenomena from the chiral brain media and the handedness of the light. For instance, classically entangled beams showed higher transmittance and constant value dependency on OAM modes than linear modes did. Also, circular polarization beam transmittance showed strong increase with topical charge OAM ( ℓ), which could be attributed to chiroptical effect.