Erik Thiel

High-resolution two-photon imaging of HE-stained samples in dermatohistopathology – A pilot study on skin tumours

Abstract Non-invasive two-photon microscopy (TPM) provides a new technique which could become in the future a substitute for hematoxylin and eosin (HE) staining of tissue sections of the epidermis and upper and middle dermis as well. High-resolution imaging, in combination with false-colour representation, allows an accurate reproduction of standard microscopy. The vertical skin viewing of epidermis and upper dermis by means of TPM allows for a new histopathological supportive technique, especially in non-melanoma skin cancer such as squamous cell carcinoma and basal cell carcinoma. If it could be used on fresh tissue samples, it could provide an alternative method to Mohs 3D histology, even though the definitive criteria for melanocytic tumours have not yet been sufficiently evaluated. Zusammenfassung Die neue, nicht-invasive Zwei-Photonen-Mikroskopie (two-photon microscopy, TPM) ermöglicht eine qualitativ hohe Reproduktion fast gleichwertig zu histologischen Präparaten exzidierter Tumoren, die die Epidermis und obere und mittlere Dermis erfassen. Mit Hilfe der durch die Scanner-Technik erzeugten hochaufgelösten Bilder in Kombination mit einer zusätzlichen Falschfarbendarstellung können relevante diagnostische wie prognostische Kriterien akkurat beurteilt werden. Die vertikale Bildgebung von Dermis und Epidermis durch die TPM ermöglicht eine ergänzende histopathologische Beurteilung vor allem bei weißem Hautkrebs wie dem Plattenepithel- oder Basalzellkarzinom. Ihr zukünftiger Einsatz an Frischpräparaten verspricht daher eine Alternative zu Mohs 3D-Histologie zu sein, auch wenn entscheidende Diagnosekriterien bei melanozytären Tumoren bislang noch nicht hinreichend beurteilbar sind.

Laser-projected photothermal thermography using thermal wave field interference for subsurface defect characterization

The coherent superposition of two anti-phased thermal wave fields creates a zone of destructive interference which is extremely sensitive to the presence of defects without any reference measurements. Combining a high power laser with a spatial light modulator allows modulating phase and amplitude of an illuminated surface that induces spatially and temporally controlled thermal wave fields. The position and depth of defects are reconstructed from analysis of the amplitude and phase of the resulting photothermal signal. The proposed concept is experimentally validated and supported by numerical modeling.

Spatial and temporal control of thermal waves by using DMDs for interference based crack detection

Active Thermography is a well-established non-destructive testing method and used to detect cracks, voids or material inhomogeneities. It is based on applying thermal energy to a samples’ surface whereas inner defects alter the nonstationary heat flow. Conventional excitation of a sample is hereby done spatially, either planar (e.g. using a lamp) or local (e.g. using a focused laser) and temporally, either pulsed or periodical. In this work we combine a high power laser with a Digital Micromirror Device (DMD) allowing us to merge all degrees of freedom to a spatially and temporally controlled heat source. This enables us to exploit the possibilities of coherent thermal wave shaping. Exciting periodically while controlling at the same time phase and amplitude of the illumination source induces – via absorption at the sample’s surface - a defined thermal wave propagation through a sample. That means thermal waves can be controlled almost like acoustical or optical waves. However, in contrast to optical or acoustical waves, thermal waves are highly damped due to the diffusive character of the thermal heat flow and therefore limited in penetration depth in relation to the achievable resolution. Nevertheless, the coherence length of thermal waves can be chosen in the mmrange for modulation frequencies below 10 Hz which is perfectly met by DMD technology. This approach gives us the opportunity to transfer known technologies from wave shaping techniques to thermography methods. We will present experiments on spatial and temporal wave shaping, demonstrating interference based crack detection.

Subsurface Defect Localization by Structured Heating Using Laser Projected Photothermal Thermography

The presented method is used to locate subsurface defects oriented perpendicularly to the surface. To achieve this, we create destructively interfering thermal wave fields that are disturbed by the defect. This effect is measured and used to locate the defect. We form the destructively interfering wave fields by using a modified projector. The original light engine of the projector is replaced with a fiber-coupled high-power diode laser. Its beam is shaped and aligned to the projectors spatial light modulator and optimized for optimal optical throughput and homogeneous projection by first characterizing the beam profile, and, second, correcting it mechanically and numerically. A high-performance infrared (IR) camera is set up according to the tight geometrical situation (including corrections of the geometrical image distortions) and the requirement to detect weak temperature oscillations at the sample surface. Data acquisition can be performed once a synchronization between the individual thermal wave field sources, the scanning stage, and the IR camera is established by using a dedicated experimental setup which needs to be tuned to the specific material being investigated. During data post-processing, the relevant information on the presence of a defect below the surface of the sample is extracted. It is retrieved from the oscillating part of the acquired thermal radiation coming from the so-called depletion line of the sample surface. The exact location of the defect is deduced from the analysis of the spatial-temporal shape of these oscillations in a final step. The method is reference-free and very sensitive to changes within the thermal wave field. So far, the method has been tested with steel samples but is applicable to different materials as well, in particular to temperature sensitive materials.

Thermal Wave Interference with High-Power VCSEL Arrays For Locating Vertically Oriented Subsurface Defects

Among the photothermal methods, full-field thermal imaging is used to characterize materials, to determine thicknesses of layers, or to find inhomogeneities such as voids or cracks. The use of classical light sources such as flash lamps (impulse heating) or halogen lamps (modulated heating) led to a variety of nondestructive testing methods, in particular, lock-in and flash-thermography. In vertical-cavity surface-emitting lasers (VCSELs), laser light is emitted perpendicularly to the surface with a symmetrical beam profile. Due to the vertical structure, they can be arranged in large arrays of many thousands of individual lasers, which allows power scaling into the kilowatt range. Recently, a high-power yet very compact version of such a VCSEL-array became available that offers both the fast timing behavior of a laser as well as the large illumination area of a lamp. Moreover, it allows a spatial and temporal control of the heating because individual parts of the array can be controlled arbitrarily in frequency, amplitude, and phase. In conjunction with a fast infrared camera, such structured heating opens up a field of novel thermal imaging and testing methods. As a first demonstration of this approach, we chose a testing problem very challenging to conventional thermal infrared testing: The detection of very thin subsurface defects perpendicularly oriented to the surface of metallic samples. First, we generate destructively interfering thermal wave fields, which are then affected by the presence of defects within their reach. It turned out that this technique allows highly sensitive detection of subsurface defects down to depths in excess of the usual thermographic rule of thumb, with no need for a reference or surface preparation.

Thermography using a 1D laser array – From planar to structured heating

Abstract In the field of optically excited thermography, flash lamps (impulse-shaped planar heating) and halogen lamps (modulated planar heating) have become established for the specific regimes of impulse and lock-in thermography. Flying-spot laser thermography is implemented by means of a rasterized focused laser, e. g. for crack detection (continuous wave operation) and photothermal material characterization (high-frequency modulated). The availability of novel technologies, i. e. fast and high-resolution IR cameras, brilliant innovative light sources and high-performance data acquisition and processing technology will enable a paradigm shift from stand-alone photothermal and thermographic techniques to uniform quantitative measurement and testing technology that is faster and more precise. Similar to an LED array, but with irradiance two orders of magnitude higher, a new type of brilliant laser source, i. e. the VCSEL array (vertical-cavity surface-emitting laser), is now available. This novel optical energy source eliminates the strong limitation to the temporal dynamics of established light sources and at the same time is spectrally clearly separated from the detection wavelength. It combines the fast temporal behavior of a diode laser with the high optical irradiance and the wide illumination area of flash lamps. In addition, heating can also be carried out in a structured manner, because individual areas of the VCSEL array can be controlled independently of each other. This new degree of freedom enables the development of completely new thermographic NDT methods.