Optische Kohärenztomographie (OCT) mit mikroskopischer Auflösung

Im Gegensatz zur konventionellen optischen Kohärenztomografie bietet die mikroskopische OCT (mOCT) eine zehnfach so hohe Auflösung, sowohl axial als auch lateral. Dadurch ist es möglich, zelluläre Morphologie in vivo darzustellen. Durch hohe Bildgebungsraten von mehreren Hundert B-Scans pro Sekunde ist es auch möglich hochdynamische Vorgänge auf zellulärer Ebene, wie etwa den Zilienschlag, zu quantifizieren.

 

Publikationen

2018

  • Hinnerk Schulz-Hildebrandt and Mario Pieper and Charlotte Stehmar and Martin Ahrens and Christian Idel and Barbara Wollenberg and Peter König and Gereon Hüttmann: Novel endoscope with increased depth of field for imaging human nasal tissue by microscopic optical coherence tomography. Biomedical Optics Express, no. 9, pp. 636-647, 2018
    BibTeX Link Link
    @article{schulz2018novel,
    title = {Novel endoscope with increased depth of field for imaging human nasal tissue by microscopic optical coherence tomography},
    author = {Hinnerk Schulz-Hildebrandt and Mario Pieper and Charlotte Stehmar and Martin Ahrens and Christian Idel and Barbara Wollenberg and Peter K\"{o}nig and Gereon H\"{u}ttmann},
    editor = {Optical Society of America},
    url = {https://www.osapublishing.org/abstract.cfm?URI=boe-9-2-636
    https://www.osapublishing.org/boe/viewmedia.cfm?uri=boe-9-2-636&seq=0},
    doi = {10.1364/BOE.9.000636},
    isbn = {10.1364/BOE.9.000636},
    year = {2018},
    date = {2018-01-16},
    journal = {Biomedical Optics Express},
    volume = {9},
    number = {2},
    pages = {636-647},
    abstract = {Intravital microscopy (IVM) offers the opportunity to visualize static and dynamic changes of tissue on a cellular level. It is a valuable tool in research and may considerably improve clinical diagnosis. In contrast to confocal and non-linear microscopy, optical coherence tomography (OCT) with microscopic resolution (mOCT) provides intrinsically cross-sectional imaging. Changing focus position is not needed, which simplifies especially endoscopic imaging. For in-vivo imaging, here we are presenting endo-microscopic OCT (emOCT). A graded-index-lens (GRIN) based 2.75 mm outer diameter rigid endoscope is providing 1.5 \textendash 2 μm nearly isotropic resolution over an extended field of depth. Spherical and chromatic aberrations are used to elongate the focus length. Simulation of the OCT image formation, suggests overall a better image quality in this range compared to a focused Gaussian beam. Total imaging depth at a reduced sensitivity and lateral resolution is more than 200 μm. Using a frame rate of 80 Hz cross-sectional images of concha nasalis were demonstrated in humans, which could resolve cilial motion, cellular structures of the epithelium, vessels and blood cells. Mucus transport velocity was determined successfully. The endoscope may be used for diagnosis and treatment control of different lung diseases like cystic fibrosis or primary ciliary dyskinesia, which manifest already at the nasal mucosa.},
    keywords = {Endoskope, OCM},
    pubstate = {published},
    tppubtype = {article}
    }
  • Hinnerk Schulz-Hildebrandt and Michael Münter and Martin Ahrens and Hendrik Spahr and Dierck Hillmann and Peter König and Gereon Hüttmann: Coherence and diffraction limited resolution in microscopic OCT by a unified approach for the correction of dispersion and aberrations. in 2nd Canterbury Conference on OCT with Emphasis on Broadband Optical Sources, no. 10591, pp. 105910O, 2018
    BibTeX Link
    @inproceedings{Schulz-Hildebrandt2018a,
    title = {Coherence and diffraction limited resolution in microscopic OCT by a unified approach for the correction of dispersion and aberrations},
    author = {Hinnerk Schulz-Hildebrandt and Michael M\"{u}nter and Martin Ahrens and Hendrik Spahr and Dierck Hillmann and Peter K\"{o}nig and Gereon H\"{u}ttmann},
    doi = {10.1117/12.2303755},
    isbn = {9781510616745},
    year = {2018},
    date = {2018-03-05},
    booktitle = {2nd Canterbury Conference on OCT with Emphasis on Broadband Optical Sources},
    volume = {10591},
    pages = {105910O},
    abstract = {Optical coherence tomography (OCT) images scattering tissues with 5 to 15 μm resolution. This is usually not sufficient for a distinction of cellular and subcellular structures. Increasing axial and lateral resolution and compensation of artifacts caused by dispersion and aberrations is required to achieve cellular and subcellular resolution. This includes defocus which limit the usable depth of field at high lateral resolution. OCT gives access the phase of the scattered light and hence correction of dispersion and aberrations is possible by numerical algorithms. Here we present a unified dispersion/aberration correction which is based on a polynomial parameterization of the phase error and an optimization of the image quality using Shannon’s entropy. For validation, a supercontinuum light sources and a costume-made spectrometer with 400 nm bandwidth were combined with a high NA microscope objective in a setup for tissue and small animal imaging. Using this setup and computation corrections, volumetric imaging at 1.5 μm resolution is possible. Cellular and near cellular resolution is demonstrated in porcine cornea and the drosophila larva, when computational correction of dispersion and aberrations is used. Due to the excellent correction of the used microscope objective, defocus was the main contribution to the aberrations. In addition, higher aberrations caused by the sample itself were successfully corrected. Dispersion and aberrations are closely related artifacts in microscopic OCT imaging. Hence they can be corrected in the same way by optimization of the image quality. This way microscopic resolution is easily achieved in OCT imaging of static biological tissues.},
    keywords = {OCM},
    pubstate = {published},
    tppubtype = {inproceedings}
    }

2017

  • Jens Horstmann and Hinnerk Schulz-Hildebrandt and Felix Bock and Sebastian Siebelmann and Eva Lankenau and Gereon Hüttmann and Philipp Steven and Claus Cursiefen: Label-Free In Vivo Imaging of Corneal Lymphatic Vessels Using Microscopic Optical Coherence Tomography. Investigative Ophthalmology & Visual Science, no. 58, pp. 5880-5886, 2017
    BibTeX Link Link
    @article{Horstmann2017,
    title = {Label-Free In Vivo Imaging of Corneal Lymphatic Vessels Using Microscopic Optical Coherence Tomography},
    author = {Jens Horstmann and Hinnerk Schulz-Hildebrandt and Felix Bock and Sebastian Siebelmann and Eva Lankenau and Gereon H\"{u}ttmann and Philipp Steven and Claus Cursiefen},
    editor = {The Association Research for in Vision and Ophthalmology},
    url = {http://iovs.arvojournals.org/article.aspx?articleid=2664271},
    doi = {10.1167/iovs.17-22286},
    isbn = {1552-5783},
    year = {2017},
    date = {2017-12-25},
    journal = {Investigative Ophthalmology & Visual Science},
    volume = {58},
    number = {13},
    pages = {5880-5886},
    abstract = {Purpose: Corneal neovascularization, in particular lymphangiogenesis, is a limiting factor in corneal transplant survival. Novel treatment approaches focus on (selective) inhibition and regression of lymphatic vessels. Imaging clinically invisible corneal lymphatic vessels is a prerequisite for these strategies. Using a murine model, this study investigates whether corneal lymphatic vessels can be imaged using microscopic optical coherence tomography (mOCT). 
    
    Methods: Corneal neovascularization was induced by intrastromal placement of 11.0 nylon sutures in one eye of BALB/c mice. After 2 weeks, cross-sectional images and volumes of the corneas with a 0.5 mm lateral and axial field of view were acquired using a custom-built mOCT system enabling a resolution of 1 μm at a B-scan rate of 165/s. Three of the six animals received an additional intrastromal injection of India ink 24 hours before the measurement to stain the corneal lymphatic system in vivo. Immunohistochemistry using CD31 and LYVE-1 was used to validate the mOCT findings. 
    
    Results: Using mOCT, lymphatic vessels were visible as dark vessel-like structures with the lumen lacking a hyperreflective wall and mostly lacking cells. However, individual, slowly moving particles, which most likely are immune cells, occasionally could be observed inside the lumen. In lymphatic vessels of ink-stained corneas, hyperreflection and shadowing underneath was observed. Ink-filled lymphatic vessels were colocalized in consecutive corneal flat mounts of the same specimen. 
    
    Conclusions: Corneal lymphatic vessels can be imaged using mOCT. This novel approach opens new options for noninvasive clinical imaging of corneal lymphatic vessels for diagnostic and therapeutic indications.},
    keywords = {OCM},
    pubstate = {published},
    tppubtype = {article}
    }

2016

  • Schulz-Hildebrandt, H. and Pieper, M. and Kasper, J. and Traulsen, N. and Mall, M. and König, P. and Hüttmann, G.: Towards automated evaluation of mucus transport measured by microscopic OCT (mOCT) during hypertonic saline treatment of Cystic Fibrosis. Pneumologie, no. 70, pp. 1-48, 2016
    BibTeX Link
    @article{Schulz-Hildebrandt2016,
       author = {Schulz-Hildebrandt, H. and Pieper, M. and Kasper, J. and Traulsen, N. and Mall, M. and König, P. and Hüttmann, G.},
       title = {Towards automated evaluation of mucus transport measured by microscopic OCT (mOCT) during hypertonic saline treatment of Cystic Fibrosis},
       journal = {Pneumologie},
       volume = {70},
       number = {07},
       pages = {1-48},
       ISSN = {0934-8387},
       DOI = {10.1055/s-0036-1584651},
       year = {2016},
       type = {Journal Article}
    }
    

2015

  • Ansari, R. and Buj, C. and Pieper, M. and Konig, P. and Schweikard, A. and Huttmann, G.: Micro-anatomical and functional assessment of ciliated epithelium in mouse trachea using optical coherence phase microscopy. Opt Express, no. 23, pp. 23217-24, 2015
    BibTeX Link
    @article{Ansari2015,
       author = {Ansari, R. and Buj, C. and Pieper, M. and Konig, P. and Schweikard, A. and Huttmann, G.},
       title = {Micro-anatomical and functional assessment of ciliated epithelium in mouse trachea using optical coherence phase microscopy},
       journal = {Opt Express},
       volume = {23},
       number = {18},
       pages = {23217-24},
       note = {1094-4087
    Ansari, Rehman
    Buj, Christian
    Pieper, Mario
    Konig, Peter
    Schweikard, Achim
    Huttmann, Gereon
    Journal Article
    United States
    Opt Express. 2015 Sep 7;23(18):23217-24. doi: 10.1364/OE.23.023217.},
       abstract = {Motile cilia perform a range of important mechanosensory and chemosensory functions, along with expulsion of mucus and inhaled pathogens from the lungs. Here we demonstrate that spectral domain optical coherence phase microscopy (SD-OCPM), which combines the principles of optical coherence tomography (OCT) and confocal microscopy, is particularly well-suited for characterization of both morphology and the ciliary dynamics of mouse trachea. We present micro-anatomical images of mouse trachea, where different cell types can be clearly visualized. The phase contrast, which measures the sub-nanometer changes in axial optical pathlength is used to determine the frequency and direction of cilia beatings.},
       ISSN = {1094-4087},
       DOI = {10.1364/oe.23.023217},
       year = {2015},
       type = {Journal Article}
    }
    

2014

  • Ansari, Rehman and Myrtus, Christian and Aherrahrou, Redouane and Erdmann, Jeanette and Schweikard, Achim and Hüttmann, Gereon: Ultrahigh-resolution, high-speed spectral domain optical coherence phase microscopy. Optics Letters, no. 39, pp. 45-47, 2014
    BibTeX Link Link
    @article{Ansari2014,
       author = {Ansari, Rehman and Myrtus, Christian and Aherrahrou, Redouane and Erdmann, Jeanette and Schweikard, Achim and Hüttmann, Gereon},
       title = {Ultrahigh-resolution, high-speed spectral domain optical coherence phase microscopy},
       journal = {Optics Letters},
       volume = {39},
       number = {1},
       pages = {45-47},
       abstract = {We present an ultrahigh-resolution, high-speed spectral domain optical coherence phase microscopy (SD-OCPM) system that combines submicrometer transverse spatial resolution and subnanometer optical path length sensitivity, with an acquisition speed of over 217,000  voxels/s. The proposed SD-OCPM system overcomes two significant drawbacks of traditional common-path interferometers—limited transverse spatial resolution and suboptimal detection sensitivity—while maintaining phase stability that is comparable with common-path interferometer setups. The transverse and axial spatial resolution of the setup is measured to be 0.6 and 1.9 μm, respectively, with a phase sensitivity of 0.0027 rad (corresponds to optical path length sensitivity of 110 pm). High-speed acquisition allows for phase-sensitive 4D imaging of biological samples with subcellular resolution.},
       keywords = {Microscopy
    Coherence imaging
    Three-dimensional microscopy},
       DOI = {10.1364/OL.39.000045},
       url = {http://ol.osa.org/abstract.cfm?URI=ol-39-1-45},
       year = {2014},
       type = {Journal Article}
    }
    

2013

  • Franke, Gesa Lilith and Hillmann, Dierck and Lührs, Christian and Koch, Peter and Wollenzin, Jörn and Hüttmann, Gereon: Towards microscopic resolution in holoscopy. pp. 85711O-85711O, 2013
    BibTeX Link Link
    @article{Franke2013,
       author = {Franke, Gesa Lilith and Hillmann, Dierck and Lührs, Christian and Koch, Peter and Wollenzin, Jörn and Hüttmann, Gereon},
       title = {Towards microscopic resolution in holoscopy},
       pages = {85711O-85711O},
       note = {10.1117/12.2006806},
       abstract = {Holoscopy is a new imaging approach combining digital holography and full-field Fourier-domain optical coherence tomography. The interference pattern between light scattered by a sample and a defined reference wave is recorded and processed numerically. During reconstruction numerical refocusing is applied, overcoming the limitation of the focal depth and thus a uniform, diffraction limited lateral resolution over the whole measurement depth can be obtained. The advantage of numerical refocusing becomes especially significant for imaging at high numerical apertures (NAs). We use a high-resolution setup based on a Mach-Zehnder interferometer with an high-resolution microscope objective (NA = 0.75). For reliable reconstruction of a sample volume the Rayleigh length of the microscope objective and the axial resolution, given by the spectral range of the light source, need to be matched. For a 0.75 NA objective a tunable light source with a sweeping range of ! 300nm is required. Here we present as a first step a tunable Ti:sapphire laser with a tuning range of 187 nm. By characterizing the spectral properties of the Ti:sapphire laser and determining the axial point spread function we demonstrate the feasibility of this light source for high-resolution holoscopy.},
       DOI = {10.1117/12.2006806},
       url = {http://dx.doi.org/10.1117/12.2006806},
       year = {2013},
       type = {Journal Article}
    }
    

  • Ansari, Rehman and Aherrahrou, Redouane and Aherrahrou, Zouhair and Erdmann, Jeanette and Huttmann, Gereon and Schweikard, Achim: Quantitative analysis of cardiomyocyte dynamics with optical coherence phase microscopy. no. 8213, pp. 821338, SPIE,
    BibTeX
    @inproceedings{Ansari2012,
       author = {Ansari, Rehman and Aherrahrou, Redouane and Aherrahrou, Zouhair and Erdmann, Jeanette and Huttmann, Gereon and Schweikard, Achim},
       title = {Quantitative analysis of cardiomyocyte dynamics with optical coherence phase microscopy},
       editor = {Joseph, A. Izatt and James, G. Fujimoto and Valery, V. Tuchin},
       publisher = {SPIE},
       volume = {8213},
       pages = {821338},
    
    }
  • Franke, Gesa Lilith and Hillmann, Dierck and Claussen, Thorsten and Luhrs, Christian and Koch, Peter and Huttmann, Gereon: High resolution holoscopy. no. 8213, pp. 821324, SPIE,
    BibTeX
    @inproceedings{Franke2012,
       author = {Franke, Gesa Lilith and Hillmann, Dierck and Claussen, Thorsten and Luhrs, Christian and Koch, Peter and Huttmann, Gereon},
       title = {High resolution holoscopy},
       editor = {Joseph, A. Izatt and James, G. Fujimoto and Valery, V. Tuchin},
       publisher = {SPIE},
       volume = {8213},
       pages = {821324},
    
    }