Microscope integrated real time 4D MHz-OCT operating at high scanning densities are capable of capturing additional visual contrast resolving depth and tissue. Even within a plain C-scan en-face projection structures are recognizable, that are not visible in a white light camera image. With advanced post processing methods, such as absorption coefficient mapping, and morphological classifiers more information is extracted. Presentation to the user in an intuitive way poses practical challenges that go beyond the implementation of a mere overlay display. We present our microscope integrated high speed 4D OCT imaging system, its clinical study use for in-vivo brain tissue imaging, and user feedback on the presentation methods we developed. In neurosurgery the de-facto standard contrast agents used for visibly highlighting brain tumors are Fluorescin and ALA, both of which come with certain caveats. As part of a clinical study we developed a microscope integrated real time 4D MHz-OCT system, operating as high scanning densities, with the intent of creating visual tissue contrast without the use of such contrast agents. Advanced post processing methods to classify tissue can be derived from static properties such as light absorption and morphology, and from dynamic properties, such as perfusion and elastography. However we also noticed that even in a plain C-scan en-face projection structures of interest could be recognized, that were not visible in the corresponding white light camera image. As part of a clinical study so far we collected data from 20 patients, used it for machine learning based classifiers and developing data presentation modalities for eventual use in a surgical environment. We present the challenges in implementing our microscope integrated high speed 4D OCT imaging system, a selection of the imaging data we collected so far during brain tumor surgeries, and the avenues toward presenting processed data to the surgeon.
In recent years, it was demonstrated that discrimination between white matter and tumor-infiltrated white matter based on optical coherence tomography (OCT) data is possible with high accuracy. However, gray matter is also present during the tumor resection and shows similar optical properties to tumor infiltration, which aggravates the tumor classification using optical coherence tomography. A semantic segmentation approach based on a convolutional neural network was applied to the problem in order to classify healthy brain tissue from tumor infiltrated brain tissue. A dataset was created, which consisted of ex vivo OCT B-scans, which were acquired by a swept-source OCT system with a central wavelength of 1300 nm. Each OCT B-scan was indirectly annotated by transforming histological labels from a corresponding H&E section onto it. The labels differentiate between white matter, gray matter and tumor infiltration. The output of the network was modeled to a Dirichlet prior distribution, which enabled the capturing of a prediction uncertainty. This approach achieved an intersection over union score of 0.72 for healthy brain tissue and 0.69 for highly tumor infiltrated brain tissue, when only confident predictions were considered.
Neuro-surgery is challenged by the difficulties of determining brain tumor boundaries during excisions. Optical coherence tomography is investigated as an imaging modality for providing a viable contrast channel. Our MHz-OCT technology enables rapid volumetric imaging, suitable for surgical workflows. We present a surgical microscope integrated MHz-OCT imaging system, which is used for the collection of in-vivo images of human brains, with the purpose of being used in machine learning systems that shall be trained to identify and classify tumorous tissue.
The ill-defined tumor borders of glioblastoma multiforme pose a major challenge for the surgeon during tumor resection, since the goal of the tumor resection is the complete removal, while saving as much healthy brain tissue as possible. In recent years, optical coherence tomography (OCT) was successfully used to classify white matter from tumor infiltrated white matter by several research groups. Motivated by these results, a dataset was created, which consisted of sets of corresponding ex vivo OCT images, which were acquired by two OCT-systems with different properties (e.g. wavelength and resolution). Each image was annotated with semantic labels. The labels differentiate between white and gray matter and three different stages of tumor infiltration. The data from both systems not only allowed a comparison of the ability of a system to identify the different tissue types present during the tumor resection, but also enable a multimodal tissue analysis evaluating corresponding OCT images of the two systems simultaneously. A convolutional neural network with dirichlet prior was trained, which allowed to capture the uncertainty of a prediction. The approach increased the sensitivity of identifying tumor infiltration from 58 % to 78 % for data with a low prediction uncertainty compared to a previous monomodal approach.
We demonstrate a 3.3 MHz A-scan rate OCT for rapid scanning of large areas of human skin. The mosaicking performance and different OCT imaging modalities including intervolume speckle contrast are evaluated.
The identification of ex vivo brain tumor tissue was investigated with two different optical coherence tomography systems exploiting two optical parameters. The optical parameters were calculated from semantically labelled OCT B-scans.
A 1.6 MHz Fourier-domain mode-locked (FDML) optical coherence tomography (OCT) was adapted to an OR-Microscope for clinical application in neurosurgery. 3D-volume scans at video rate are envisaged with approximately 50μm lateral and 20μm axial resolution
In Fourier domain mode locked (FDML) lasers, extremely precise and stable matching of the filter tuning period and light circulation time in the cavity is essential for ultra-low noise operation. During the operation of FDML lasers, the ultra-low noise mode can be lost due to temperature drifts of the already temperature stabilized cavity resulting in increased intensity noise. Until now, the filter frequency is continuously regulated to match the changing light circulation time. However, this causes the filter frequency to constantly change by a few mHz and leads to synchronization issues in cases where a fixed filter frequency is desired. We present an actively cavity length controlled FDML laser and a robust and high precision feedback loop algorithm for maintaining ultra-low noise operation. Instead of adapting the filter frequency, the cavity length is adjusted by a motorized free space beam path to match the fixed filter frequency. The closed-loop system achieves a stability of ~0.18 mHz at a sweep repetition rate of ~418 kHz which corresponds to a ratio of 4×10-10. We investigate the coherence properties during the active cavity length adjustments and observe no noise increase compared to fixed cavity length. The cavity length control is fully functional and for the first time, offers the possibility to operate an FDML laser in sweet spot mode at a fixed frequency or phase locked to an external clock. This opens new possibilities for system integration of FDML lasers.
Optical coherence tomography (OCT) has the potential to become an additional imaging modality for surgical guidance in the field of neurosurgery, especially when it comes to the detection of different infiltration grades of glioblastoma multiforme at the tumor border. Interpretation of the images, however, is still a big challenge. A method to create a labeled OCT dataset based on ex vivo brain samples is introduced. The tissue samples were embedded in an agarose mold giving them a distinctive shape before images were acquired with two OCT systems (spectral domain (SD) and swept source (SS) OCT) and histological sections were created and segmented by a neuropathologist. Based on the given shape, the corresponding OCT images for each histological image can be determined. The transfer of the labels from the histological images onto the OCT images was done with a non-affine image registration approach based on the tissue shape. It was demonstrated that finding OCT images of a tissue sample corresponding to segmented histological images without any color or laser marking is possible. It was also shown that the set labels can be transferred onto OCT images. The accuracy of method is 26 ± 11 pixel, which translates to 192 ± 75 μm for the SS-OCT and 94 ± 43 μm for the SD-OCT. The dataset consists of several hundred labeled OCT images, which can be used to train a classification algorithm.
The routine pathology workflow relies on cutting tissue into single-cell layer thick slices using paraffin or frozen sectioning. We propose a fast method to obtain sections of equivalent quality optically using the strong sectioning capabilities of two-photon microscopy (TPM). Hematoxylin and eosin (HE) equivalent staining of the tissue is achieved using acridine orange and sulforhodamine 101. We improved our previously presented pulsed fiber laser to deliver adjustable pulse durations of ~30ps at repetition rates of up to 16MHz and kW peak power. We can now image up to one square centimeter of tissue with sub-micrometer resolution within 15 minutes.
Fourier domain mode locked (FDML) laser are fast swept light sources. Measuring the linewidth and coherence length of such light sources is not straightforward, but very important for a physical understanding of FDML lasers and their performance in optical coherence tomography (OCT). In order to characterize the dynamic (“instantaneous”) linewidth, we performed beat signal measurements between a stationary narrowband continuous wave laser and an FDML laser and detected the signals with a 63 GHz real time oscilloscope. The evaluation of the beat signals of consecutive FDML wavelength sweeps yields information about the phase evolution within one sweep and over several sweeps. These measurements suggest the existence of a distinct comb like mode structure of the FDML laser and help to determine the locking strength of individual modes (comb lines).
The separation of tumorous brain tissue and healthy brain tissue is still a big challenge in the field of neurosurgery, especially when it comes to the detection of different infiltration grades of glioblastoma multiforme at the tumor border. On the basis of a recently created labelled OCT dataset of ex vivo glioblastoma multiforme tumor samples the detection of brain tumor tissue and the identification of zones with varying degrees of infiltration of tumor cells was investigated. The identification was based on the optical properties, which were extracted by an exponential fit function. The results showed that a separation of tumorous tissue and healthy white matter based on these optical properties is possible. A support vector machine was trained on the optical properties to separate tumor from healthy white matter tissue, which achieved a sensitivity of 91% and a specificity of 76% on an independent training dataset.
The aim of this work is the creation of segmented data set consisting of optical coherence tomography (OCT) scans, which were taken of brain tumor tissue with different tumor infiltration rates. In an ongoing clinical study more than 140 human brain samples with different infiltration grades were recorded ex vivo with two OCT systems, a spectral domain OCT system and a swept-source OCT system that uses a 1310 nm Fourier domain mode locked laser. The histological analysis of the recorded samples builds the ground truth for labeling the corresponding OCT B-Scans. The segmented data set gained from this process will be used to train a classification algorithm, taking into account structural and optical properties such as the attenuation coefficient. In the future the classification algorithm together with a microscope integrated OCT system will be used for the in vivo identification of brain tumors as a guidance tool for the surgeon to increase tumor resection efficiency.
Fourier domain mode locking (FDML) is a recently developed technique for lasers to generate ultra-rapid wavelength sweeps, equivalent to a train of extremely chirped pulses. FDML lasers are the light sources of choice for fastest megahertz optical coherence tomography (MHz-OCT). Measuring the coherence properties of FDML lasers is of particular importance for the image quality in OCT but it is also crucial to develop a better understanding of this unconventional mode locking mechanism. Usually, experiments to analyze the phase stability of FDML lasers use interferometers to generate interference of a single laser by delaying a part of the output to generate a beat signal. Here, for the first time, we present real time beat signal measurements between two independent FDML lasers over the entire sweep range of ~5 THz width for more than 80 roundtrips (~200 μs), evaluate their phase stability and explain the consequence for our understanding of the FDML mechanism. Beat signal measurements allow direct access to the phase difference between the FDML lasers and therefore the difference in timing of the circulating sweeps as well as their instantaneous frequency.
Optical coherence tomography (OCT) applications like ultra-widefield and full eye-length imaging are of high interest for various diagnostic purposes. In swept-source OCT these techniques require a swept light source, which is coherent over the whole imaging depth. We present a zero roll-off 1060 nm Fourier Domain Mode Locked-Laser (FDML-Laser) for retinal OCT imaging at 1.7 MHz A-scan rate and first long-range imaging results with it. Several steps such as improved dispersion compensation and frequency regulation were performed and will be discussed. Besides virtually no loss in OCT signal over the maximum depth range of 4.6 mm and very good dynamic range was observed. Roll-off measurements show no decrease of the point-spread function (PSF), while maintaining a high dynamic range.
We implemented a real-time video-rate 4D-OCT system with virtual reality display. To achieve the required low latencies we optimized the dataflow path and the placement of the necessary synchronization points. Employing temporal reprojection enables to perform volume rendering at 1/3 of the display refresh rate, yet maintaining smooth updates to the HMD; thus we achieve display updates at 90Hz, volume rendering at 30Hz and C-scan acquisition at <15Hz. By mounting of a tracking accessory to the scanning head we can render the OCT volume in virtual space in the position of the actual imaging volume.
Optical coherence tomography (OCT) is a non-invasive imaging technique which is currently investigated for intraoperative detection of residual tumor during resection of human gliomas. Three different OCT systems were used for imaging of human glioblastoma in vivo (830nm spectral domain (SD) OCT integrated into a surgical microscope) and ex vivo (940nm SD-OCT and 1310nm swept-source MHz-OCT using a Fourier domain mode locked (FDML) laser). Before clinical data acquisition, the systems were characterized using a three-dimensional point-spread function phantom. To distinguish tumor from healthy brain tissue later on, attenuation coefficients of each pixel in OCT depth profiles are calculated. First examples from a clinical study show that the pixel-resolved calculation of the attenuation coefficient provides a good image contrast and confirm that white matter shows a higher signal and more homogeneous signal structure than tumorous tissue.
MHz-OCT systems based on FDML swept laser sources combined with the massive parallel processing capabilities of modern computer hardware enable volumetric imaging, processing and stereoscopic display at video rates. The increasing image quality and speed might enable new fields of application where the volumetric OCT completely replaces stereoscopic microscopes instead of being a mere supplement. Aside from the depth resolving capability, a particular advantage is the ability to display a whole image volume from arbitrary points of view without the need to move the actual microscope or to rotate the patient’s eye. Purely digital microscopy is already offered as alternative to traditional through-an-eyepiece surgical microscope. We explore the use of virtual reality to present digital OCT microscopy images to a trained surgeon, carrying out a series of surgical procedures ex-vivo on a porcine eye model.
Two-Photon Microscopy (TPM) can provide three-dimensional morphological and functional contrast in vivo. Through proper staining, TPM can be utilized to create virtual, HE equivalent images and thus can improve throughput in histology-based applications. We previously reported on a new light source for TPM that employs a compact and robust fiber-amplified, directly modulated laser. This laser is pulse-to-pulse wavelength switchable between 1064 nm, 1122 nm, and 1186 nm with an adjustable pulse duration from 50ps to 5ns and arbitrary repetition rates up to 1MHz at kW-peak powers. Despite the longer pulse duration, it can achieve similar average signal levels compared to fs-setups by lowering the repetition rate to achieve similar cw and peak power levels. The longer pulses lead to a larger number of photons per pulse, which yields single shot fluorescence lifetime measurements (FLIM) by applying a fast 4 GSamples/s digitizer. In the previous setup, the wavelengths were limited to 1064 nm and longer. Here, we use four wave mixing in a non-linear photonic crystal fiber to expand the wavelength range down to 940 nm. This wavelength is highly suitable for imaging green fluorescent proteins in neurosciences and stains such as acridine orange (AO), eosin yellow (EY) and sulforhodamine 101 (SR101) used for histology applications. In a more compact setup, we also show virtual HE histological imaging using a direct 1030 nm fiber MOPA.
The absorption of nanosecond laser pulses induces rapid thermo-elastic deformation in tissue. A sub-micrometer scale displacement occurs within a few microseconds after the pulse arrival. We investigate the thermo-elastic deformation using a 1.5 MHz phase-sensitive optical coherence tomography (OCT) system. An analysis of the results shows that the displacement is dominated by the optical absorption. By tuning the excitation wavelength, thermo-elastic displacement spectrum can be extracted, showing the similar features as optical absorption spectrum. By choosing proper excitation wavelength, targeted tissue type can be highlighted, which further enables a new imaging modality, so called thermo-elastic OCT.
We demonstrate that the 3.2 MHz a-scan rate and the improved coherence of our new low noise FDML laser enables live 3D-OCT with different spectral zooms and up to 10 cm of imaging range.
FDML lasers provide sweep rates in the MHz range at wide optical bandwidths, making them ideal sources for high
speed OCT. Recently, at lower speed, ultralong-range swept-source OCT has been demonstrated using a tunable
vertical cavity surface emitting laser (VCSEL) and also using a Vernier-tunable laser. These sources provide relatively
high sweep rates and meter range coherence lengths. In order to achieve similar coherence, we developed an extremely
well dispersion compensated Fourier Domain Mode Locked (FDML) laser, running at 3.2 MHz sweep rate and 120 nm
spectral bandwidth. We demonstrate that this laser offers meter range coherence and enables volumetric long range OCT
of moving objects.
Absorption of nanosecond laser pulses induces rapid thermo-elastic deformation in tissue, i.e. a sub-micrometer scale displacement happens within a couple of microseconds. In this study, we initially investigate the depth-resolved deformation using a 1.5 MHz phase-sensitive optical coherence tomography (OCT) system. Functional images can be reconstructed based on the detected deformation, which enables a new imaging modality called thermo-elastic deformation imaging (TDI). Our results show that the associated displacement is related to the optical absorption of the short laser pulses. The TDI images can provide tissue type information in addition to the conventional OCT images.
Over the last 20 years, optical coherence tomography (OCT) has become a valuable diagnostic tool in ophthalmology with several 10,000 devices sold today. Other applications, like intravascular OCT in cardiology and gastro-intestinal imaging will follow. OCT provides 3-dimensional image data with microscopic resolution of biological tissue in vivo. In most applications, off-line processing of the acquired OCT-data is sufficient. However, for OCT applications like OCT aided surgical microscopes, for functional OCT imaging of tissue after a stimulus, or for interactive endoscopy an OCT engine capable of acquiring, processing and displaying large and high quality 3D OCT data sets at video rate is highly desired.
We developed such a prototype OCT engine and demonstrate live OCT with 25 volumes per second at a size of 320x320x320 pixels. The computer processing load of more than 1.5 TFLOPS was handled by a GTX 690 graphics processing unit with more than 3000 stream processors operating in parallel. In the talk, we will describe the optics and electronics hardware as well as the software of the system in detail and analyze current limitations. The talk also focuses on new OCT applications, where such a system improves diagnosis and monitoring of medical procedures. The additional acquisition of hyperspectral stimulated Raman signals with the system will be discussed.
We present full volumetric high speed OCT imaging of the retina with multiple settings varying in volume size and volume rate. The volume size ranges from 255x255 A-scans to 160x40 A-scans with 450 samples per depth scan with volume rates varying between 20.8 V/s for the largest volumes to 195.2 V/s for the smallest. The system is based on a 1060nm Fourier domain mode locked (FDML) laser with 1.6MHz line rate. Scanning along the fast axis is performed with a 2.7 kHz or 4.3 kHz resonant scanner operated in bidirectional scanning mode, while a standard galvo scanner is used for the slow axis. The performance is analyzed with respect to various potential applications, like intraoperative OCT.
While FDML lasers with MHz sweep speeds have been presented five years ago, these devices have required manual control for startup and operation. Here, we present a fully self-starting and continuously regulated FDML laser with a sweep rate of 1.5 MHz. The laser operates over a sweep range of 115 nm centered at 1315 nm, and provides very high average output power of more than 100 mW. We characterize the laser performance, roll-off, coherence length and investigate the wavelength and phase stability of the laser output under changing environmental conditions. The high output power allows optical coherence tomography (OCT) imaging with an OCT sensitivity of 108 dB at 1.5 MHz.
Full volumetric high speed OCT imaging of the retina with multiple settings varying in volume size and volume rate is
presented. The volume size ranges from 255x255 A-scans to 160x40 A-scans with 450 samples per depth scan. The
volume rates vary between 20.8 V/s for the largest volumes to 195.2 V/s for the smallest. The system is based on a
1060nm Fourier domain mode locked (FDML) laser with 1.6MHz line rate. Scanning along the fast axis is performed
with a 2.7 kHz or 4.3 kHz resonant scanner operated in bidirectional scanning mode, while a standard galvo scanner is
used for the slow axis. The performance is analyzed with respect to various potential applications, like intraoperative
OCT.
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