Bessel light beam for a surgical laser focusing telescope-a novel approach.

As the demand for CO[Formula: see text] laser surgeries continues to grow, the quality of their main instrument, the laser micromanipulator, becomes increasingly important. However, in many surgery systems, a large ratio of the laser power is wasted due to the reflection from the mirror of a telescopic system, like a Cassegrain telescope, back to the laser side, which not only decreases the system's efficiency but can also damage the system itself. In this article, we introduce a new design of the micromanipulator telescope for CO[Formula: see text] laser surgery, which employs a Bessel beam to improve the system efficiency. As in the propagation of a Bessel beam, the power of the light beam can be transferred from the center to a ring shape, the whole power reflected from the first mirror can reach the second mirror and no power goes back to the second mirror hole. The micromanipulator telescope design and optimization are carried out using Zemax Optics Studio, and the integration of the Bessel beam into the system is implemented using MATLAB. Our simulation results show that by applying the appropriate Bessel beam, the system efficiency can reach more than 96%, and the normalized peak irradiance can increase by 40 to 73% for various working distances. In addition to increasing the system efficiency and normalized peak irradiance, resulting in a sharper surgical blade, the use of the Bessel beam enhances the depth of focus, making the system less sensitive to depth misalignment.

Evaluation of Raman spectroscopic macro raster scans of native cervical cone biopsies using histopathological mapping.

Raman spectroscopy based discrimination of cervical precancer and normal tissue has been shown previously in vivo with fiber probe based measurements of colposcopically selected sites. With a view to developing in vivo large area imaging, macro raster scans of native cervical cone biopsies with an average of 200 spectra per sample are implemented (n=16). The diagnostic performance is evaluated using histopathological mapping of the cervix surface. Different data reduction and classification methods (principal component analysis, wavelets, k-nearest neighbors, logistic regression, partial least squares discriminant analysis) are compared. Using bootstrapping to estimate confidence intervals for sensitivity and specificity, it is concluded that differences among different spectra classification procedures are not significant. The classification performance is evaluated depending on the tissue pathologies included in the analysis using the average performance of different classification procedures. The highest sensitivity (91%) and specificity (81%) is obtained for the discrimination of normal squamous epithelium and high-grade precancer. When other non-high-grade tissue sites, such as columnar epithelium, metaplasia, and inflammation, are included, the diagnostic performance decreases.

Stable and tunable single frequency Nd:GSAG laser around 943 nm.

Stable single frequency output around 943 nm was obtained from a quasi-continuous wave (qcw) diode-pumped, Q-switched Nd:GSAG laser. The Q-switched Nd:GSAG laser was injection seeded with a single mode laser diode. Its frequency was stabilized by an active-control loop specially designed for a strong qcw pump. The spectral linewidth of the Nd:GASG laser was 41 MHz and the frequency stability was 10 MHz. The single-frequency-pulsed laser generated 32 mJ pulse energy at 10 Hz repetition rate. When the repetition rate was increased to 100 Hz, 5.6 mJ pulse energy was obtained by a thermal dynamic stable resonator. By tuning the seed laser, the wavelength of the pulsed Nd:GASG laser can be continuously varied from 942.1 to 943.1 nm.

Operation of a Raman laser in bulk silicon.

A Raman laser based on a bulk silicon single crystal with 1.127 µm emission wavelength is demonstrated. The Si crystal with 30 mm length was placed into an external cavity and pumped by a Q-switched Nd:YAG master oscillator power amplifier system. Strong defocusing of the pump and Raman laser beam by free carriers was compensated by an intracavity lens. Raman laser operation with a pulse duration of 2.5 ns was identified by a Raman laser threshold significantly lower than the single-pass stimulated Raman-scattering threshold. Linear absorption losses of the 1.06415 µm pump radiation are strongly reduced by cooling the Si crystal to a temperature of 10 K.

Functional domain size in aggregates of light-harvesting complex II and thylakoid membranes.

The functional domain size for efficient excited singlet state quenching was studied in artificial aggregates of the main light-harvesting complex II (LHCIIb) from spinach and in native thylakoid membranes by picosecond time-resolved fluorescence spectroscopy and quantum yield measurements. The domain size was estimated from the efficiency of added exogenous singlet excitation quenchers-phenyl-p-benzoquinone (PPQ) and dinitrobenzene (DNB). The mean fluorescence lifetimes τ(av) were quantified for a range of quencher concentrations. Applying the Stern-Volmer formalism, apparent quenching rate constants k(q) were determined from the dependencies on quencher concentration of the ratio τ(0)(av)/τ(av), where τ(0)(av) is the average fluorescence lifetime of the sample without addition of an exogenous quencher. The functional domain size was gathered from the ratio k(q)'/k(q), i.e., the apparent quenching rate constants determined in aggregates (or membranes), k(q)', and in detergent-solubilised LHCII trimers, k(q), respectively. In LHCII macroaggregates, the resulting values for the domain size were 15-30 LHCII trimers. In native thylakoid membranes the domain size was equivalent to 12-24 LHCII trimers, corresponding to 500-1000 chlorophylls. Virtually the same results were obtained when membranes were suspended in buffers promoting either membrane stacking or destacking. These domain sizes are orders of magnitude smaller than the number of physically connected pigment-protein complexes. Therefore our results imply that the physical size of an antenna system beyond the numbers of a functional domain size has little or no effect on improving the light-harvesting efficiency.

Excitation energy transfer in intact cells and in the phycobiliprotein antennae of the chlorophyll d containing cyanobacterium Acaryochloris marina.

The cyanobacterium Acaryochloris marina is unique because it mainly contains Chlorophyll d (Chl d) in the core complexes of PS I and PS II instead of the usually dominant Chl a. Furthermore, its light harvesting system has a structure also different from other cyanobacteria. It has both, a membrane-internal chlorophyll containing antenna and a membrane-external phycobiliprotein (PBP) complex. The first one binds Chl d and is structurally analogous to CP43. The latter one has a rod-like structure consisting of three phycocyanin (PC) homohexamers and one heterohexamer containing PC and allophycocyanin (APC). In this paper, we give an overview on the investigations of excitation energy transfer (EET) in this PBP-light-harvesting system and of charge separation in the photosystem II (PS II) reaction center of A. marina performed at the Technische Universität Berlin. Due to the unique structure of the PBP antenna in A. marina, this EET occurs on a much shorter overall time scale than in other cyanobacteria. We also briefly discuss the question of the pigment composition in the reaction center (RC) of PS II and the nature of the primary donor of the PS II RC.

Quantitative Raman spectroscopy in turbid media.

Intrinsic Raman spectra of biological tissue are distorted by the influences of tissue absorption and scattering, which significantly challenge signal quantification. A combined Raman and spatially resolved reflectance setup is introduced to measure the absorption coefficient micro(a) and the reduced scattering coefficient micro(s) (') of the tissue, together with the Raman signals. The influence of micro(a) and micro(s) (') on the resonance Raman signal of beta-carotene is measured at 1524 cm(-1) by tissue phantom measurements and Monte Carlo simulations for micro(a)=0.01 to 10 mm(-1) and micro(s) (')=0.1 to 10 mm(-1). Both methods show that the Raman signal drops roughly proportional to 1 micro(a) for micro(a)>0.2 mm(-1) in the measurement geometry and that the influence of micro(s) (') is weaker, but not negligible. Possible correction functions dependent on the elastic diffuse reflectance are investigated to correct the Raman signal for the influence of micro(a) and micro(s) ('), provided that micro(a) and micro(s) (') are measured as well. A correction function based on the Monte Carlo simulation of Raman signals is suggested as an alternative. Both approaches strongly reduce the turbidity-induced variation of the Raman signals and allow absolute Raman scattering coefficients to be determined.

Single frequency operation of a tunable injection-seeded Nd:GSAG Q-switched laser around 942nm.

Single frequency operation of a diode-pumped tunable injection-seeded Nd:GSAG Q-switched laser around 942nm was demonstrated. With a three-mirror ring cavity, the single frequency laser pulse with output energy of 13.2mJ was obtained at a repetition rate of 10Hz. The linewidth of the single frequency laser was less than 100MHz. The wavelength of the single frequency Nd:GSAG laser can be tuned from 942.38nm to 943.10nm.

Excitation energy transfer from phycobiliprotein to chlorophyll d in intact cells of Acaryochloris marina studied by time- and wavelength-resolved fluorescence spectroscopy.

The fluorescence decay spectra and the excitation energy transfer from the phycobiliproteins (PBP) to the chlorophyll-antennae of intact cells of the chlorophyll (Chl) d-dominated cyanobacterium Acaryochloris marina were investigated at 298 and 77 K by time- and wavelength-correlated single photon counting fluorescence spectroscopy. At 298 K it was found that (i) the fluorescence dynamics in A. marina is characterized by two emission peaks located at about 650 and 725 nm, (ii) the intensity of the 650 nm fluorescence depends strongly on the excitation wavelength, being high upon excitation of phycobiliprotein (PBP) at 632 nm but virtually absent upon excitation of chlorophyll at 430 nm, (iii) the 650 nm fluorescence band decayed predominantly with a lifetime of 70 +/- 20 ps, (iv) the 725 nm fluorescence, which was observed independent of the excitation wavelength, can be described by a three-exponential decay kinetics with lifetimes depending on the open or the closed state (F(0) or F(m)) of the reaction centre of Photosystem II (PS II). Based on the results of this study, it is inferred that the excitation energy transfer from phycobiliproteins to Chl d of PS II in A. marina occurs with a time constant of about 70 ps, which is about three times faster than the energy transfer from the phycobilisomes to PS II in the Chl a-containing cyanobacterium Synechococcus 6301. A similar fast PBP to Chl d excitation energy transfer was also observed at 77 K. At 77 K a small long-lived fluorescence decay component with a lifetime of 14 ns was observed in the 640-700 nm spectral range. However, it has a rather featureless spectrum, not typical for Chl a, and was only observed upon excitation at 400 nm but not upon excitation at 632 and 654 nm. Thus, this long-lived fluorescence component cannot be used as an indicator that the primary PS II donor of Acaryochloris marina contains Chl a.