Biocompatible Materials Analytical Tools PDF

376-1714-00L Biocompatible Materials Analytical tools, microscopy, bioMEMS, patterning 20.11.2024 Prof. Dr. Katharina Maniura, Empa, Biointerfaces/ D-HEST Dr. Markus Rottmar, Empa, Biointerfaces Prof. Dr. Marcy Zenobi-Wong, ETHZ, D-HEST, Tissue Engineering & Fabrication Basic principle in (bio)sen...

376-1714-00L Biocompatible Materials Analytical tools, microscopy, bioMEMS, patterning 20.11.2024 Prof. Dr. Katharina Maniura, Empa, Biointerfaces/ D-HEST Dr. Markus Rottmar, Empa, Biointerfaces Prof. Dr. Marcy Zenobi-Wong, ETHZ, D-HEST, Tissue Engineering & Fabrication Basic principle in (bio)sensing ▪ ▪ November 20, 2024 Katharina Maniura 2 Analysis methods – recap lecture protein adsorption November 20, 2024 Katharina Maniura 3 Ellipsometry November 20, 2024 Katharina Maniura 4 Optical Waveguide Lightmode Spectroscopy (OWLS) The OWLS technique measures refractive index changes of which the adsorbed mass can be calculated using de Feijter‘s formula. Since this formula only contains the difference between the refractive indices of the adsorbed layer and the cover medium, the coupled water is not visible by this technique. November 20, 2024 Katharina Maniura 5 Quartz Crystal Microbalance (QCM) - 1 DOI:10.5772/intechopen.97576 November 20, 2024 Katharina Maniura 6 Quartz Crystal Microbalance (QCM) - 2 A Quartz Crystal Microbalance Immunosensor for Stem Cell Selection and Extraction Sensors 2017, 17(12), 2747; https://doi.org/10.3390/s17122747 November 20, 2024 Katharina Maniura 7  Biotin-streptavidin conjugation system Illustration of the Streptavidin-Biotin interaction. The multivalent properties of streptavidin allow it to bind up to four biotin molecules with a high degree of affinity. Biotin is typically conjugated to an enzyme, antibody, or target protein https://www.aatbio.com/catalog/biotin-and-streptavidin November 20, 2024 Katharina Maniura 8  Protein/ biomolecule adsorption analysis techniques ▪ various techniques available to study protein / cell – biomaterial interactions ▪ label free vs. label detection methods ▪ in situ vs. ex situ techniques ▪ combinatorial sensing and chemical mapping techniques are required to understand the dynamic arrangement and conformation of proteins, cells and tissues on (bio)material surfaces ▪ in situ sensing of the build-up of hierarchical strutures allows to understand the effect of surface engineering for the host response November 20, 2024 Katharina Maniura 9  Light microscopy – conjugate planes ▪ The imaging and illumination ray paths through a microscope adjusted for Köhler illumination are presented, with the focal conjugates of each plane set indicated by crossover points of the ray traces. ▪ Illustrated diagrammatically in the figure is the reciprocal nature of the two sets of conjugate planes that occur in the microscope. ▪ The optical relationship between the conjugate plane sets is based upon the fact that, in the illuminating ray path (shown in red), the spherical wave fronts converge and are brought into focus onto the aperture planes, while in the imaging ray path (shown in yellow), the spherical waves converge into focused rays in the field planes. ▪ Light rays that are focused in one set of conjugate planes are nearly parallel when passing through the other set of conjugate planes. ▪ The reciprocal relationship between the two sets of conjugate planes determines how the two ray paths fundamentally interact in forming an image in the microscope, and it also has practical consequences for operation of the microscope. https://www.microscopyu.com/microscopy-basics/conjugate-planes-in-optical-microscopy November 20, 2024 Katharina Maniura 10  Light microscopy - contrast https://micro.magnet.fsu.edu/primer/techniques/contrast.html November 20, 2024 Katharina Maniura 11 One sample: 4 different ways to look at it November 20, 2024 Katharina Maniura 12 Fluorescence microscopy in biomaterial research November 20, 2024 Katharina Maniura 13 Confocal microscopy: 3D sectioning of specimen November 20, 2024 Katharina Maniura 14 Diffraction and resolution in microscopy November 20, 2024 Katharina Maniura 15 Point Spread Function of a point source of light Airy disc = descriptions of the best- focused spot of light that a perfect lens with a circular aperture can make, limited by the diffraction of light November 20, 2024 Katharina Maniura 16 Convolution ▪ Image formation in a confocal microscope: central longitudinal (XZ) slice. ▪ The 3D acquired distribution arises from the convolution of the real light sources with the PSF. November 20, 2024 Katharina Maniura 17 Deconvolution November 20, 2024 Katharina Maniura 18 Super-resolution microscopy: imaging beyond the diffraction limit Structured Illumination Microscopy November 20, 2024 Katharina Maniura 19 PALM / STORM – photoactivated localization microscopy November 20, 2024 Katharina Maniura 20 PALM / STORM examples (2D) November 20, 2024 Katharina Maniura 21 PALM / STORM examples (3D) November 20, 2024 Katharina Maniura 22 Quantitative (fluorescence) microscopy – word of caution November 20, 2024 Katharina Maniura 23  Histological analysis November 20, 2024 Katharina Maniura 24 Electron microscopy in biomaterial research check out courses at ScopeM! November 20, 2024 Katharina Maniura 25 Sample preparation for EM TEM November 20, 2024 Katharina Maniura 26 Focused ion beam (FIB) – SEM (1) November 20, 2024 Katharina Maniura 27 Focused ion beam (FIB) – SEM (2) The response of soft tissue cells to Ti implants is modulated by blood- implant interactions Lackington et al., Materials Today Bio 15 (2022) 100303 November 20, 2024 Katharina Maniura 28  (Bio)MEMS for mechanotransduction studies November 20, 2024 Katharina Maniura 29  Mechanotransduction tools November 20, 2024 Katharina Maniura 30  Vasculature – shear flow November 20, 2024 Katharina Maniura 31 November 20, 2024 Katharina Maniura 32 November 20, 2024 Katharina Maniura 33 November 20, 2024 Katharina Maniura 34 Stretchable transwell chamber platform November 20, 2024 Katharina Maniura 35 Stretchable transwell chamber platform November 20, 2024 Katharina Maniura 36 Uniaxial stretching of epithelial monolayers November 20, 2024 Katharina Maniura 37 Encoded FRET biosensors – force sensing and remodeling of E-cadherin at cell-cell junctions November 20, 2024 Katharina Maniura 38 Force transduction at cadherin cell-cell junctions November 20, 2024 Katharina Maniura 39 Microfabricated strain array for cell and microtissue mechanotransduction studies November 20, 2024 Katharina Maniura 40 Integrin-specific molecular FRET tension sensors November 20, 2024 Katharina Maniura 41 Spatial patterning of cells and tissues November 20, 2024 Katharina Maniura 42 Microcontact printing of proteins to pattern cells November 20, 2024 Katharina Maniura 43 Challenges with microcontact printing November 20, 2024 Katharina Maniura 44 Lift-off protein patterning to control cell shape November 20, 2024 Katharina Maniura 45 Summary microscopy, bioMEMS, patterning November 20, 2024 Katharina Maniura 46