Stefan Kray

Simultaneous dual-band ultra-high resolution optical coherence tomography

Ultra-high resolution optical coherence tomography (OCT) imaging is demonstrated simultaneously at 840 nm and 1230 nm central wavelength using an off-the-shelf turn-key supercontinuum light source. Spectral filtering of the light source emission results in a double peak spectrum with average powers exceeding 100 mW and bandwidths exceeding 200 nm for each wavelength band. A free-space OCT setup optimized to support both wavelengths in parallel is introduced. OCT imaging of biological tissue ex vivo and in vivo is demonstrated with axial resolutions measured to be < 2 mum and < 4 mum at 840 nm and 1230 nm, respectively. This measuring scheme is used to extract spectroscopic features with outstanding spatial resolution enabling enhanced image contrast.

High-resolution simultaneous dual-band spectral domain optical coherence tomography

We present a fiber-based spectral-domain optical coherence tomography system, measuring simultaneously at 740 nm and 1300 nm central wavelengths. Real-time imaging is demonstrated with axial resolutions <3 μm and <5 μm, respectively. This technique allows for in vivo functional OCT imaging with high spatial resolution and outstanding spectroscopic imaging contrast.

Electronically controlled coherent linear optical sampling for optical coherence tomography

Electronically controlled coherent linear optical sampling for low coherence interferometry (LCI) and optical coherence tomography (OCT) is demonstrated, using two turn-key commercial mode-locked fiber lasers with synchronized repetition rates. This novel technique prevents repetition rate limitations present in previous implementations based on asynchronous optical sampling. Adjustable scanning ranges and scanning rates are realized within an interferometric setup by full electronic control of the mutual time delay of the two laser pulse trains. We implement this novel linear optical sampling scheme with broad spectral bandwidths for LCI, optical filter characterization and OCT imaging in two and three dimensions.

Optical Tomography of MMP Activity Allows a Sensitive Noninvasive Characterization of the Invasiveness and Angiogenesis of SCC Xenografts

For improved tumor staging and therapy control, imaging biomarkers are of great interest allowing a noninvasive characterization of invasiveness. In squamous epithelial skin and cervix lesions, transition to invasive stages is associated with enhanced matrix metalloproteinase (MMP) activity, increased angiogenesis, and worsened prognosis. Thus, we investigated MMP activity as imaging biomarker of invasiveness and the potential of optical tomography in characterizing the angiogenic and invasive behavior of skin squamous cell carcinoma (SCC) xenografts. MMP activity was measured in vivo in HaCaT-ras A-5RT3 tumors at different angiogenic and invasive stages (onset of angiogenesis, intermediate and highly angiogenic, invasive stage) and after 1 week of sunitinib treatment by fluorescence molecular tomography-microcomputed tomography imaging using an activatable probe. Treatment response was additionally assessed morphologically by optical coherence tomography (OCT). In vivo MMP activity significantly differed between the groups, revealing highest levels in the highly angiogenic, invasive tumors that were confirmed by immunohistochemistry. At the onset of angiogenesis with lowest MMP activity, fibroblasts were detected in the MMP-positive areas, whereas macrophages were absent. Accumulation of both cell types occurred in both invasive groups, again to a significantly higher degree at the most invasive and angiogenic stage. Sunitinib treatment significantly reduced the MMP activity and accumulation of fibroblasts and macrophages and blocked tumor invasion that was additionally visualized by OCT. Human cervical SCCs also showed high MMP activity and a similar stromal composition as the HaCaT xenografts, whereas normal tissue was negative. This study strongly suggests MMP activity as imaging biomarker and demonstrates the high sensitivity of optical tomography in determining tumor invasiveness that can morphologically be supported by OCT.

Increased tissue contrast by high resolution simultaneous dual-band optical coherence tomography in three dimensions

Enhanced visualization of tissue contrast and morphological boundaries is demonstrated by analyzing OCT volume data in two distinct wavelength regions. The extension of this so-called simultaneous dual-band method to three dimensions is realized by a custom-built 3D spectral domain OCT system imaging in the 800 und 1200 nm wavelength domain. Color representations of the spectral differences of high resolution OCT volume data significantly simplify the discrimination of different tissue structures beyond the capabilities of cross-sectional spectroscopic OCT approaches.

The Ex vivo Eye Irritation Test (EVEIT) model as a mean of improving venom ophthalmia understanding

&NA; Snakes belonging to the genus Naja (Elapid family), also known as “spitting cobras”, can spit venom towards the eyes of the predator as a defensive strategy, causing painful and potentially blinding ocular envenoming. Venom ophthalmia is characterized by pain, hyperemia, blepharitis, blepharospasm and corneal erosions. Elapid venom ophthalmia is not well documented and no specific treatment exists. Furthermore, accidental ejection of venom by non‐spitting vipers, as Bothrops, also occurs. The Ex vivo Eye Irritation Test model (EVEIT) has enabled important progress in the knowledge of chemical ocular burns. Considering the lack of experimental animal model, we adapted the EVEIT to study venom ophthalmia mechanisms. Ex vivo rabbit corneas were exposed to venoms from spitting (Naja mossambica, Naja nigricollis) and non‐spitting (Naja naja, Bothrops jararaca and Bothrops lanceolatus) snakes, and rinsed or not with water. The corneal thickness and the depth of damage were assessed using high‐resolution optical coherence tomography (HR‐OCT) imaging and histological analysis. All Naja venoms induced significant corneal edema, collagen structure disorganization and epithelial necrosis. Corneas envenomed by African N. mossambica and N. nigricollis venoms were completely opaque. Opacification was not observed in corneas treated with venoms from non‐spitting snakes, such as the Asian cobra, N. naja, and the vipers, B. jararaca and B. lanceolatus. Moreover, Bothrops venoms were able to damage the epithelium and cause collagen structure disorganization, but not edema. Immediate water rinsing improved corneal status, though damage and edema could still be observed. In conclusion, the present study shows that the EVEIT model was successfully adapted to set a new experimental ex vivo animal model of ophthalmia, caused by snake venoms, which will enable to explore new therapies for venom ophthalmia. HighlightsEx vivo Eye Irritation Test (EVEIT) was successfully adapted to set a new ex vivo animal model of snake venom ophthalmia.The EVEIT model reproduces the corneal opacification observed in case of African Elapid venom ophthalmia.Improvement of the corneal status, after envenomation, could be achieved by immediate water decontamination.

Surface profilometry and low coherence interferometry by electronically controlled optical sampling

In optical sampling, a transient waveform under study is sampled with time delayed laser pulses, achieving a high time resolution. The limitations in measurement time due to the mechanical adjustment of the time delay can be overcome by asynchronous optical sampling (ASOPS) [1], where two pulsed lasers with slightly detuned repetition rates are employed. This technique is commonly used for studying transient characteristics [1], for infrared spectrometry [2] as well as remote sensing applications [3] and optical coherence tomography [4]. However, a static difference in repetition rates leads to static scanning ranges, resulting in unnecessary large measurement times and large amounts of measured data especially for the latter applications.

Pathogenesis of the dry eye syndrome observed by optical coherence tomography in vitro

Three dimensional optical coherence tomography (OCT) is introduced as a valuable tool to analyze the pathogenesis of corneal diseases. Here, OCT in combination with a novel in vitro model for the dry eye syndrome enables an improved understanding of the underlying damaging process of the ocular surface. En-face OCT projections indicate a deep structural damage of the epithelium and anterior stroma by osmotic forces.

Supercontinuum Based Ultrahigh Resolution Fourier Domain Optical Coherence Tomography for in vivo Dermatology

A Fourier domain optical coherence tomography system is demonstrated, employing a commercial supercontinuum light source for the 1300nm wavelength region. Ultrahigh axial resolution of 1mm and 100dB sensitivity are achieved.