Two-layer reconstruction of Raman spectra in diffusive media based on an analytical model in the time domain.

We derive and validate an analytical model that describes the migration of Raman scattered photons in two-layer diffusive media, based on the diffusion equation in the time domain. The model is derived under a heuristic approximation that background optical properties are identical on the excitation and Raman emission wavelengths. Methods for the reconstruction of two-layer Raman spectra have been developed, tested in computer simulations and validated on tissue-mimicking phantom measurements data.

Perspectives on interstitial photodynamic therapy for malignant tumors.

Significance: Despite remarkable advances in the core modalities used in combating cancer, malignant diseases remain the second largest cause of death globally. Interstitial photodynamic therapy (IPDT) has emerged as an alternative approach for the treatment of solid tumors.

Aim: The aim of our study is to outline the advancements in IPDT in recent years and provide our vision for the inclusion of IPDT in standard-of-care (SoC) treatment guidelines of specific malignant diseases.

Computer simulation analysis of source-detector position for percutaneously measured O -gas signal in a three-dimensional preterm infant lung.

Further improvements in the clinical care of our most vulnerable patients-preterm infants-are needed. Novel diagnostic and surveillance tools facilitate such advances. The GASMAS technique has shown potential to become a tool to, noninvasively, monitor gas in the lungs of preterm infants, by placing a laser source and a detector on the chest wall skin.

Development of a 3-dimensional tissue lung phantom of a preterm infant for optical measurements of oxygen-Laser-detector position considerations.

There is a need to further improve the clinical care of our most vulnerable patients-preterm infants. Novel diagnostic and treatment tools facilitate such advances. Here, we evaluate a potential percutaneous optical monitoring tool to assess the oxygen and water vapor content in the lungs of preterm babies.

Bioreactor scale up and protein product quality characterization of piggyBac transposon derived CHO pools.

Chinese hamster ovary (CHO) cells remain the most popular host for the production of biopharmaceutical drugs, particularly monoclonal antibodies (mAbs), bispecific antibodies, and Fc-fusion proteins. Creating and characterizing the stable CHO clonally-derived cell lines (CDCLs) needed to manufacture these therapeutic proteins is a lengthy and laborious process. Therefore, CHO pools have increasingly been used to rapidly produce protein to support and enable preclinical drug development.

Online dosimetry for temoporfin-mediated interstitial photodynamic therapy using the canine prostate as model.

Online light dosimetry with real-time feedback was applied for temoporfin-mediated interstitial photodynamic therapy (PDT) of dog prostate. The aim was to investigate the performance of online dosimetry by studying the correlation between light dose plans and the tissue response, i.e.

Alkaline cation-exchange chromatography for the reduction of aggregate and a mis-formed disulfide variant in a bispecific antibody purification process.

During the purification development of a bispecific antibody, cation-exchange chromatography was screened for its ability to separate a prominently expressed (>12%) mis-formed disulfide bond variant, termed MAb-diabody, and aggregate from the product of interest. The influence of pH, product load (g of product per liter of resin) and linear velocity on the separations were evaluated for the strong cation-exchange resins SP Sepharose HP and POROS(®) HS50. Cation-exchange chromatography is commonly operated distant to the isoelectric point of a molecule, generally leading to acidic conditions for antibody purification.

System for interstitial photodynamic therapy with online dosimetry: first clinical experiences of prostate cancer.

The first results from a clinical study for Temoporfin-mediated photodynamic therapy (PDT) of low-grade (T1c) primary prostate cancer using online dosimetry are presented. Dosimetric feedback in real time was applied, for the first time to our knowledge, in interstitial photodynamic therapy. The dosimetry software IDOSE provided dose plans, including optical fiber positions and light doses based on 3-D tissue models generated from ultrasound images.

In vivo photosensitizer tomography inside the human prostate.

Interstitial photodynamic therapy (IPDT) provides a promising means to treat large cancerous tumors and solid organs inside the human body. The treatment outcome is dependent on the distributions of light, photosensitizer, and tissue oxygenation. We present a scheme for reconstructing the spatial distribution of a fluorescent photosensitizer.

Fluorescence intensity and lifetime imaging of free and micellar-encapsulated doxorubicin in living cells.

Frequency domain fluorescence lifetime imaging microscopy (FLIM) has been used in combination with laser scanning confocal microscopy to study the cellular uptake behavior of the antitumor drug doxorubicin (DOX) and micellar-encapsulated DOX (PLyAd-DOX). The endocytosis uptake process of PLyAd-DOX was monitored over 72 hours using confocal microscopy, with a maximum fluorescence recorded at incubation periods around 24 hours. The micellar structure was not found to release the encapsulated DOX during the time course of imaging.

Realtime light dosimetry software tools for interstitial photodynamic therapy of the human prostate.

Photodynamic therapy (PDT) for the treatment of prostate cancer has been demonstrated to be a safe treatment option capable of inducing tissue destruction and decreasing prostate specific antigen (PSA) levels. However, prostate-PDT results in large intra- and interpatient variations in treatment response, possibly due to biological variations in tissue composition and short-term response to the therapeutic irradiation. Within our group, an instrument for interstitial PDT on prostate tissue has been developed that combines therapeutic light delivery and monitoring of light transmission via numerous bare-ended optical fibers.

A white light confocal microscope for spectrally resolved multidimensional imaging.

Spectrofluorometric imaging microscopy is demonstrated in a confocal microscope using a supercontinuum laser as an excitation source and a custom-built prism spectrometer for detection. This microscope system provides confocal imaging with spectrally resolved fluorescence excitation and detection from 450 to 700 nm. The supercontinuum laser provides a broad spectrum light source and is coupled with an acousto-optic tunable filter to provide continuously tunable fluorescence excitation with a 1-nm bandwidth.

Calibration of a wide-field frequency-domain fluorescence lifetime microscopy system using light emitting diodes as light sources.

High brightness light emitting diodes are an inexpensive and versatile light source for wide-field frequency-domain fluorescence lifetime imaging microscopy. In this paper a full calibration of an LED based fluorescence lifetime imaging microscopy system is presented for the first time. A radio-frequency generator was used for simultaneous modulation of light emitting diode (LED) intensity and the gain of an intensified charge coupled device (CCD) camera.

Feasibility of white-light time-resolved optical mammography.

We demonstrate the feasibility of white-light time-resolved optical mammography. The instrumentation is based on supercontinuum light generated in photonic crystal fiber and 32-channel parallel time-correlated single-photon-counting detection. Total measurement time is of the order of 10 min for typical clinical applications.

Application of frequency-domain Fluorescence Lifetime Imaging Microscopy as a quantitative analytical tool for microfluidic devices.

We describe the application of wide-field frequency domain Fluorescence Lifetime Imaging Microscopy (FLIM) to imaging in microfluidic devices. FLIM is performed using low cost, intensity modulated Light Emitting Diodes (LEDs) for illumination. The use of lifetime imaging for quantitative analysis within such devices is demonstrated by mapping the molecular diffusion of iodide ions across a microchannel.

Time-resolved spectrally constrained method for the quantification of chromophore concentrations and scattering parameters in diffusing media.

We have devised and experimentally validated, on tissue-simulating phantoms and in vivo, a time-resolved spectral fitting analysis for direct assessment of chromophore concentrations and scattering parameters. Experimental data have been acquired with a time-resolved broadband system based on supercontinuum light generated in a photonic crystal fiber and a 32 channel Time Correlated Single Photon Counting system. The novel method is more robust than conventional techniques, especially at low signal-to-noise ratio.

Dynamic time-resolved diffuse spectroscopy based on supercontinuum light pulses.

We present a detailed characterization of a system for fast time-resolved spectroscopy of turbid media based on supercontinuum generation in a photonic crystal fiber. The light source provides subpicosecond pulses in the 550-1000-nm spectral range, at 85 MHz, at an average power of up to 50 mW. Wavelength-resolved detection is accomplished by means of a spectrometer coupled to a 16-channel, multianode photomultiplier tube, giving a resolution of 4.

Characterization of normal breast tissue heterogeneity using time-resolved near-infrared spectroscopy.

In recent years, extensive efforts have been made in developing near-infrared optical techniques to be used in detection and diagnosis of breast cancer. Variations in optical properties of normal breast tissue set limits to the performance of such techniques and must therefore be thoroughly examined. In this paper, we present intra- and intersubject as well as contralateral variations of optical and physiological properties in breast tissue as measured by using four-wavelength time-resolved spectroscopy (at 660, 786, 916 and 974 nm).

Performance assessment of photon migration instruments: the MEDPHOT protocol.

We propose a comprehensive protocol for the performance assessment of photon migration instruments. The protocol has been developed within the European Thematic Network MEDPHOT (optical methods for medical diagnosis and monitoring of diseases) and is based on five criteria: accuracy, linearity, noise, stability, and reproducibility. This protocol was applied to a total of 8 instruments with a set of 32 phantoms, covering a wide range of optical properties.

Fluorescence spectra provide information on the depth of fluorescent lesions in tissue.

The fluorescence spectrum measured from a fluorophore in tissue is affected by the absorption and scattering properties of the tissue, as well as by the measurement geometry. We analyze this effect with Monte Carlo simulations and by measurements on phantoms. The spectral changes can be used to estimate the depth of a fluorescent lesion embedded in the tissue by measurement of the fluorescence signal in different wavelength bands.

Numerical simulations of light scattering by red blood cells.

Scattering of electromagnetic waves from a red blood cell is simulated using the finite-difference time-domain method (FDTD), the Rytov approximation and the discrete dipole approximation (DDA). Both FDTD and DDA are full wave methods that give accurate results in a wide range of wavelengths. The Rytov approximation is a much simpler method that is limited to scattering angles within 30 degrees from the forward direction.

Spectroscopic time-resolved diffuse reflectance and transmittance measurements of the female breast at different interfiber distances.

The first, to our knowledge, in-vivo broadband spectroscopic characterization of breast tissue using different interfiber distances as well as transmittance measurements is presented. Absorption and scattering properties are measured on six healthy subjects, using time-resolved diffuse spectroscopy and an inverse model based on the diffusion equation. Wavelength-tunable picosecond-pulse lasers and time-correlated single-photon counting detection are employed, enabling fully spectroscopic measurements in the range 610 to 1040 nm.

Time-resolved spectrophotometer for turbid media based on supercontinuum generation in a photonic crystal fiber.

We describe an instrument for the time-resolved spectroscopy of turbid media that is based on supercontinuum generation in a photonic crystal fiber. The light injected into the sample consists of subpicosecond pulses that cover 550-1000 nm at 85 MHz at an average power of as much as 40 mW. A spectrometer coupled to a multianode photomultiplier tube is used to detect the light simultaneously in 16 wavelength channels, with a resolution of 5-20 nm/channel, depending on the grating.

Light scattering by multiple red blood cells.

The interaction of light with multiple red blood cells was systematically investigated by the finite-difference time-domain method (FDTD). The simulations showed that the lateral multiple scattering between red blood cells is very weak and that the polarization has an almost insignificant influence on the distribution of the scattered light. The numerical results of the FDTD method were compared with the results from the Rytov approximation and the discrete dipole approximation (DDA).