Portable System for In-Clinic Differentiation of Skin Cancers from Benign Skin Lesions and Inflammatory Dermatoses.

The exquisite sensitivity of Raman spectroscopy for detecting biomolecular changes in skin cancer has previously been explored; however, this mostly required analysis of excised tissue samples using bulky, immobile laboratory instrumentation. In this study, the technique was translated for clinical use with a portable Raman system and customized fiber optic probe and applied to differentiation of skin cancers from benign lesions and inflammatory dermatoses. The aim was to provide an easy-to-use, easy-to-manage assessment tool for clinicians to use in their daily patient examination routine to perform rapid Raman measurements of skin lesions in vivo. Using this system, >867 spectra were measured in vivo from 330 patients with a wide variety of different benign skin lesions (n = 603), inflammatory dermatoses (n = 140), and skin cancers (n = 124). Ethnicities represented were 70% European; 16% Asian; 6% Maori; 5% Pacific people; and 4% Middle East, Latin American, and African. Accurate differentiation of skin cancers from benign lesions and inflammatory dermatoses was achieved using partial least squares discriminant analysis, with area under curve for the receiver operator curves for external validation sets ranging from 0.916 to 0.958. This study shows evidence for robust clinical translation of Raman spectroscopy for rapid, accurate diagnosis of skin cancer.

Raman spectroscopy and mass spectrometry identifies a unique group of epidermal lipids in active discoid lupus erythematosus.

Discoid lupus erythematosus (DLE) is the most common form of cutaneous lupus1. It can cause permanent scarring. The pathophysiology of is not fully understood. Plasmacytoid dendritic cells are found in close association with apoptotic keratinocytes inferring close cellular signalling. Matrix Associated Laser Desorption Ionisation (MALDI) combined with Fourier Transform Ion Cyclotron Resonance Mass Spectrometry (FT-ICR-MS) is an exquisitely sensitive combination to examine disease processes at the cellular and molecular level. Active areas of discoid lupus erythematosus were compared with normal perilesional skin using MALDI combined with FT-ICR-MS. A unique set of biomarkers, including epidermal lipids is identified in active discoid lupus. These were assigned as sphingomyelins, phospholipids and ceramides. Additionally, increased levels of proteins from the keratin, and small proline rich family, and aromatic amino acids (tryptophan, phenylalanine, and tyrosine) in the epidermis are observed. These techniques, applied to punch biopsies of the skin, have shown a distinctive lipid profile of active discoid lupus. This profile may indicate specific lipid signalling pathways. Lipid rich microdomains (known as lipid rafts) are involved in cell signalling and lipid abnormalities have been described with systemic lupus erythematosus which correlate with disease activity.

Raman spectroscopy system for real-time diagnosis of clinically significant prostate cancer tissue.

Prostate cancer (PCa) is a significant healthcare problem worldwide. Current diagnosis and treatment methods are limited by a lack of precise in vivo tissue analysis methods. Real-time cancer identification and grading could dramatically improve current protocols. Here, we report the testing of a thin optical probe using Raman spectroscopy (RS) and classification methods to detect and grade PCa accurately in real-time. We present the first clinical trial on fresh ex vivo biopsy cores from an 84 patient cohort. Findings from 2395 spectra measured on 599 biopsy cores show high accuracy for diagnosing and grading PCa. We can detect clinically significant PCa from benign and clinically insignificant PCa with 90% sensitivity and 80.2% specificity. We also demonstrate the ability to differentiate cancer grades with 90% sensitivity and specificity ≥82.8%. This work demonstrates the utility of RS for real-time PCa detection and grading during routine transrectal biopsy appointments.

Classification of Preeclamptic Placental Extracellular Vesicles Using Femtosecond Laser Fabricated Nanoplasmonic Sensors.

Placental extracellular vesicles (EVs) play an essential role in pregnancy by protecting and transporting diverse biomolecules that aid in fetomaternal communication. However, in preeclampsia, they have also been implicated in contributing to disease progression. Despite their potential clinical value, current technologies cannot provide a rapid and effective means of differentiating between healthy and diseased placental EVs. To address this, a fabrication process called laser-induced nanostructuring of SERS-active thin films (LINST) was developed to produce scalable nanoplasmonic substrates that provide exceptional Raman signal enhancement and allow the biochemical fingerprinting of EVs. After validating the performance of LINST substrates with chemical standards, placental EVs from tissue explant cultures were characterized, demonstrating that preeclamptic and normotensive placental EVs have classifiably distinct Raman spectra following the application of advanced machine learning algorithms. Given the abundance of placental EVs in maternal circulation, these findings encourage immediate exploration of surface-enhanced Raman spectroscopy (SERS) of EVs as a promising method for preeclampsia liquid biopsies, while this novel fabrication process will provide a versatile and scalable substrate for many other SERS applications.

Raman spectroscopy reveals age- and sex-related differences in cortical bone from people with osteoarthritis.

Bone strength in human cortical bone is determined by the composition and structure of both the mineral and collagen matrices and influenced by factors such as age, gender, health, lifestyle and genetic factors. Age-related changes in the bone matrix are known to result in loss of mechanical strength and increased fragility. In this study we show how Raman spectroscopy, with its exquisite sensitivity to the molecular structure of bone, reveals new insights into age- and sex-related differences. Raman analysis of 18 samples of cortical hip bone obtained from people aged between 47-82 years with osteoarthritis (OA) found subtle changes in the lipid and collagen secondary structure, and the carbonate (CO32-) and phosphate (PO43-) mineral ratios in the bone matrix. Significant differences were observed between older and younger bones, and between older female and older male bones; no significant differences were observed between younger male and female bones. Older female bones presented the lowest mineral to matrix ratios (MMR) and highest CO32-/PO43- ratios, and relative to lipid/collagen -CH2 deformation modes at 1450 cm-1 they had lowest overall mineral content, higher collagen cross linking and lipid content but lower levels of α-helix collagen structures than older male and younger male and female bones. These observations provided further insight on bone composition changes observed in the bone volume fraction (BV/TV) for the older female bones from microCT measurements on the same samples, while tissue mineral density (TMD) measurements had shown no significant differences between the samples.

Femtosecond lasers for high-precision orthopedic surgery.

Laser micromachining with ultrashort pulses has shown great promise for clean, safe surgical treatment of bone tissue. However, comparisons of performance and development of "best practice" have been hampered by the difficulty of comparing results across a wide variety of experimental approaches and under surgically irrelevant conditions (e.g., dried, dead bone). Using a femtosecond (fs) pulsed laser system (τ = 140 fs, repetition rate = 1 kHz, λ = 800 nm), a comprehensive study of femtosecond laser microsurgery using the standard metrics of laser micromachining (ablation threshold, incubation effects, ablation rates, effect of focal point depth within the material and heat affected zone (HAZ)) was conducted on live, freshly harvested bovine and ovine cortical bone. Three important points of optimism for future implementation in the surgical theatre were identified: (1) the removal of material is relatively insensitive to the focal point depth within the material, removing the need for extreme depth precision for excellent performance; (2) femtosecond laser ablation of fresh bone demonstrates very little incubation effect, such that multiple passes of the laser over the same region of bone removes the same amount of material; and (3) the complete absence of collateral damage, heat- or shock-induced, on both the macro- and microscopic scales can be achieved readily, within a broad parameter range. Taken together, these results indicate a handheld or robotic deployed fiber laser platform for femtosecond laser microsurgery is a very viable prospect.

Raman rogue waves in a partially mode-locked fiber laser.

We report on an experimental study of spectral fluctuations induced by intracavity Raman conversion in a passively partially mode-locked, all-normal dispersion fiber laser. Specifically, we use dispersive Fourier transformation to measure single-shot spectra of Raman-induced noise-like pulses, demonstrating that for low cavity gain values Raman emission is sporadic and follows rogue-wave-like probability distributions, while a saturated regime with Gaussian statistics is obtained for high pump powers. Our experiments further reveal intracavity rogue waves originating from cascaded Raman dynamics.

Coherence and shot-to-shot spectral fluctuations in noise-like ultrafast fiber lasers.

We report on experimental studies of coherence and fluctuations in noise-like pulse trains generated by ultrafast fiber oscillators. By measuring the degree of first-order coherence using a Young's-type interference experiment, we prove the lack of phase coherence across the seemingly regular array of pulses. We further quantify the pulse-to-pulse fluctuations by recording the single-shot spectra of the megahertz pulse train, and experimentally demonstrate the existence of spectral fluctuations that remain unresolved in conventional time-averaged ensemble measurements. Phase incoherence and spectral fluctuations are contrasted with quantified coherence and spectral stability when the laser is soliton mode-locked.

Raman-driven destabilization of mode-locked long cavity fiber lasers: fundamental limitations to energy scalability.

We report on the destabilization of the mode-locking operation of a long cavity fiber laser. We show that the destabilization is accompanied by the abrupt emergence of a strong frequency-downshifted Stokes signal, and simultaneously, we find that the laser output displays characteristics typical of noise-like pulses. We use numerical simulations to illustrate how the Stokes signal grows from stimulated Raman scattering and plays a key role in the destabilization of the laser output. Our results indicate that stimulated Raman scattering may impose an ultimate limit on the energy scalability via cavity lengthening.

Environmentally stable all-PM all-fiber giant chirp oscillator.

We report on an environmentally stable giant chirp oscillator operating at 1030 nm. Thanks to the use of a nonlinear amplifying loop mirror as the mode-locker, we are able to extract pulse energies in excess of 10 nJ from a robust all-PM cavity with no free-space elements. Extensive numerical simulations reveal that the output oscillator energy and duration can simply be up-scaled through the lengthening of the cavity with suitably positioned single-mode fiber. Experimentally, using different cavity lengths we have achieved environmentally stable mode-locking at 10, 3.7 and 1.7 MHz with corresponding pulse energies of 2.3, 10 and 16 nJ. In all cases external grating-pair compression below 400 fs has been demonstrated.

Mode-locked femtosecond all-normal all-PM Yb-doped fiber laser using a nonlinear amplifying loop mirror.

We report on a new design for a passively mode locked fibre laser employing all normal dispersion polarisation maintaining fibres operating at 1 µm. The laser produces linearly polarized, linearly chirped pulses that can be recompressed down to 344 fs. Compared to previous laser designs the cavity is mode-locked using a nonlinear amplifying fibre loop mirror that provides an additional degree of freedom allowing easy control over the pulse parameters. This is a robust laser design with excellent reliability and lifetime.

Parabolic and hyper-Gaussian similaritons in fiber amplifiers and lasers with gain saturation.

We present a new asymptotically exact analytical similariton solution of the generalized nonlinear Schrdinger equation for pulses propagating in fiber amplifiers and lasers with normal dispersion including the effect of gain saturation. Numerical simulations are in excellent agreement with this analytical solution describing self-similar linearly chirped parabolic pulses. We have also found that for small enough values of the dimensionless saturation energy parameter the fiber amplifiers and lasers can generate a new type of linearly chirped self-similar pulses, which we call Hyper-Gaussian similaritons. The analytical Hyper-Gaussian similariton solution of the generalized nonlinear Schrdinger equation is also in a good agreement with numerical simulations.

Quasi-parabolic pulse propagation and breakup in fiber amplifiers with third-order dispersion.

We present an analytical solution for propagating pulses in normal dispersion fiber amplifiers, including the effect of third-order dispersion. The solution of the generalized nonlinear Schrödinger equation is based on asymptotic methods, first-order perturbation theory, and a renormalization procedure and leads to determination of the critical length corresponding to pulse breakup. We have also found and confirmed numerically the condition on the parameters that govern the propagation, as is necessary to ensure a highly accurate analytical description of the pulses and critical lengths in fiber amplifiers with third-order dispersion.

Experimental realization of a mode-locked parabolic Raman fiber oscillator.

We report here the first demonstration of a mode-locked fiber laser delivering parabolic pulses (similaritons) at 1534 nm. The use of a Raman-based gain medium potentially allows its implementation at any wavelength. The 22nJ output similariton pulses have a true parabolic shape both in the time and spectral domains and a linear chirp. Linear recompression close to Fourier limit is demonstrated allowing us to obtain 6 ps compressed pulses with a compression factor of 75.