Resonance polarization and phase-mismatched CARS of pheophytin b excited in the Qy band

Resonance polarization and phase-mismatched coherent anti-Stokes Raman scattering (CARS) measurements were performed on pheophytin b dissolved in acetone excited in the Qy absorption band, where strong broad fluorescence makes spontaneous Raman spectroscopy impossible. The phase-mismatching technique was applied to suppress solvent background and used in combination with the polarization-sensitive CARS technique to measure directly the x1111(3) and x1221(3) components to estimate depolarization ratios. The spectra were fitted by a non-linear least-squares procedure yielding vibrational band parameters. Some CARS dispersion information on the vibrational amplitudes was obtained by varying the pump wavelength. CARS excitation profiles based on transform theory were calculated and partly explain the observed amplitude dispersion. The application of the combined phase-mismatched polarization CARS technique may be useful in many other cases of highly fluorescing molecules when resonantly excited

Possibilities and limitations of off-resonance polarization sensitive cars of short chain proteins

Polarization sensitive CARS in the absence of resonance enhancement is applied to a short chain protein. The minimum concentration to record polarization sensitive CARS spectra of protein solutions is estimated to be 10 mg/ml. The effects limiting the protein concentration are discussed and regarded from an experimental point of view. Signal strength and line parameters of polarization sensitive CARS spectra of the short chain protein Lysyl-Tryptophyl-Lysine are compared with those of a normal Raman spectrum

Non-resonant background suppression in preresonance CARS spectra of flavin adenine dinucleotide: Demonstration of a background suppression technique using phase mismatching and comparison with the polarization-sensitive CARS technique

Polarization-sensitive CARS spectra of a 5.7 × 10-3 mol dm-3 flavin adenine dinucleotide (FAD) solution were recorded under preresonance conditions at a pump wavelength of 532 nm. The depolarization ratios of the vibrations are shown to be close to the depolarization ratio of the non-resonant background. This results in a severe reduction of the vibration resonant signal (a factor of 700-900) in the polarization CARS spectrum, and a poor improvement in the ratio of the resonant signal and the non-resonant background (<10). \ud In this context, a non-resonant background suppression technique is discussed and demonstrated for 5.7 × 10-3 and 1.4 × 10-3 mol dm-3 FAD solutions excited at 532 nm; the non-resonant susceptibility of the walls of the cuvette, which contains the FAD solution, is used to compensate the non-resonant signal contribution of the solution. An improvement in the signal-to-noise ratio of ca. 50 is achieved at the cost of a factor of 30 in the resonant signal strength. Lorentzian-shaped spectral bands are obtained, facilitating the determination of band position, width and intensity. Line shape parameters and depolarization ratios for FAD are extracted from the presented spectra by curve fitting. The signal strength and background suppression achieved with these techniques and the resonance CARS technique (at a pump wavelength of 480 nm) are compared and discussed

Optical spectroscopy: current advances and future applications in cancer diagnostics and therapy

Optical spectroscopy (OS) is a tissue-sensing technique that could enhance cancer diagnosis and treatment in the near future. With OS, tissue is illuminated with a selected light spectrum. Different tissue types can be distinguished from each other based on specific changes in the reflected light spectrum that are a result of differences on a molecular level between compared tissues. Therefore, OS has the potential to become an important optical tool for cancer diagnosis and treatment monitoring. In recent years, significant progress has been made in the discriminating abilities of OS techniques between normal and cancer tissues of multiple human tissue types. This article provides an overview of the advances made with diffuse reflectance, fluorescence and Raman spectroscopy techniques in the field of clinical oncology, and focuses on the different clinical applications that OS could enhance

Modelling the assessment of port wine stain parameters from skin surface temperature following a diagnostic laser pulse

Laser treatment of port wine stains (PWS) has become an established clinical modality over the past decade. However, in some cases full clearance of the PWS cannot be achieved. To improve the clinical results, it is necessary to match the laser treatment parameters to the PWS anatomy on an individual patient basis. Therefore, knowledge of the PWS structure is of great importance. The objective of this study is to describe a diagnostic method to assess the PWS blood vessels depth and diameter from the skin surface temperature-time course following a diagnostic laser pulse. The Monte Carlo (MC) method was used to calculate the deposited laser energy into a port wine stain skin model following irradiation by a diagnostic laser pulse at 577 nm. The heat equation was solved numerically, using the deposited energy profile as the source term, yielding the temperature-time course at the skin surface. Subtraction of "bloodless" skin signal from that of the skin containing blood vessels gives us the net contribution of a heated dermal blood vessel to the skin surface temperature-time behaviour. The net blood vessel signal shows heat-diffusion behaviour and was found to be sensitive to the dermal blood vessel depth and diameter. The time delay for the peak signal temperature to occur depends quadratically on the blood vessel depth. The peak temperature relates linearly to the blood vessel diameter. The degree of epidermal melanin content can also be determined from the immediate temperature rise of the signal. The proposed method easily enables assessment of the blood vessel depth and diameter as well as the epidermal melanin content in a skin model. The method can be applied to a real PWS when using the adjacent normal skin as a referenc