Proceedings ArticleDOI
Two-photon laser scanning microscopy on native cartilage and collagen-membranes for tissue-engineering
TLDR
In these experiments 2-Photon laser scanning microscopy has been used to acquire 3-dimensional structural information on native unstained biological samples for tissue engineering purposes and imaging depth, fluorescence intensity and surface topology appear promising as key information for discriminating between the non-arthritic and arthritic states.Abstract:
In our experiments 2-Photon laser scanning microscopy (2PLSM) has been used to acquire 3-dimensional structural information on native unstained biological samples for tissue engineering purposes. Using near infrared (NIR) femtosecond laser pulses for 2-photon excitation and second harmonic generation (SHG) it was possible to achieve microscopic images at great depths in strongly (light) scattering collagen membranes (depth up to 300 μm) and cartilage samples (depth up to 460 μm). With the objective of optimizing the process of chondrocyte growth on collagen scaffolding materials for implantation into human knee joints, two types of samples have been investigated. (1) Both arthritic and non-arthritic bovine and human cartilage samples were examined in order to differentiate between these states and to estimate the density of chondrocytes. In particular, imaging depth, fluorescence intensity and surface topology appear promising as key information for discriminating between the non-arthritic and arthritic states. Human chondrocyte densities between 2-10 6 /cm 3 and 20-10 6 /cm 3 , depending on the relative position of the sample under investigation within the cartilage, were measured using an automated procedure. (2) Chondrocytes which had been sown out on different types of I/III-collagen membranes, were discriminated from the scaffolding membranes on the basis of their native fluorescence emission spectra. With respect to the different membranes, either SHG signals from the collagen fibers of the membranes or differences in the emission spectra of the chondrocytes and the scaffolding collagenes were used to identify chondrocytes and membranes.read more
Citations
More filters
Journal ArticleDOI
Two-Photon Polymerization for Microfabrication of Three-Dimensional Scaffolds for Tissue Engineering Application
TL;DR: Three-dimensional microfabrication using two-photon polymerization of biocompatible photopolymers is applied for producing scaffolds for tissue engineering of artificial cartilage using methacrylated oligo(lactide-co-caprolactone), urethanes, polyglycerines and biopolymers.
Journal ArticleDOI
Visualizing Osteogenesis In Vivo Within a Cell–Scaffold Construct for Bone Tissue Engineering Using Two-Photon Microscopy
TL;DR: Visualizing the in vivo progression of the cell and scaffold microenvironment will contribute to the understanding of tissue-engineered regeneration and should lead to the development of more streamlined and therapeutically powerful approaches.
Journal ArticleDOI
Two-photon techniques in tissue engineering.
TL;DR: It was able to show that spectral autofluorescence imaging provides spatially resolved data for the non-invasive online control of the tissue engineering process as well as the quantification of cell distribution within the scaffold.
Journal ArticleDOI
Multifocal two-photon laser scanning microscopy combined with photo-activatable GFP for in vivo monitoring of intracellular protein dynamics in real time
TL;DR: The intracellular dynamics of the Arabidopsis MYB transcription factor LHY/CCA1-like 1 (LCL1) that contains both a nuclear import and a nuclear export signal was quantitatively investigated and an export-negative mutant of LCL1 remained trapped inside the nucleus.
Book ChapterDOI
Systems nanobiology: from quantitative single molecule biophysics to microfluidic-based single cell analysis.
Joerg Martini,Wibke Hellmich,Dominik Greif,Anke Becker,Thomas Merkle,Robert Ros,Alexandra Ros,Katja Toensing,Dario Anselmetti +8 more
TL;DR: This chapter presents three methodological applications that demonstrate how quantitative informations can be accessed that are representative for cellular processes or single cell analysis like gene expression regulation, intracellular protein translocation dynamics, and single cell protein fingerprinting.
References
More filters
Journal ArticleDOI
Two-Photon Laser Scanning Fluorescence Microscopy
TL;DR: The fluorescence emission increased quadratically with the excitation intensity so that fluorescence and photo-bleaching were confined to the vicinity of the focal plane as expected for cooperative two-photon excitation.
Journal ArticleDOI
Nonlinear magic: multiphoton microscopy in the biosciences
TL;DR: Multiphoton microscopy has found a niche in the world of biological imaging as the best noninvasive means of fluorescence microscopy in tissue explants and living animals and its use is now increasing exponentially.
Journal ArticleDOI
A review of the optical properties of biological tissues
TL;DR: The known optical properties (absorption, scattering, total attenuation, effective attenuation and/or anisotropy coefficients) of various biological tissues at a variety of wavelengths are reviewed in this article.
Journal ArticleDOI
Articular cartilage repair: basic science and clinical progress. A review of the current status and prospects.
TL;DR: The existence of many new and encouraging biological approaches to cartilage repair justifies the future investment of time and money in this research area, particularly given the extremely high socio-economic importance of such therapeutic strategies in the prevention and treatment of these common joint diseases and traumas.
Journal ArticleDOI
Live tissue intrinsic emission microscopy using multiphoton-excited native fluorescence and second harmonic generation
Warren R. Zipfel,Rebecca M. Williams,R. H. Christie,Alexander Yu. Nikitin,Bradley T. Hyman,Watt W. Webb +5 more
TL;DR: Applications involving a range of intrinsic molecules and molecular assemblies that enable direct visualization of tissue morphology, cell metabolism, and disease states such as Alzheimer's disease and cancer are compiled and demonstrated.