The intrinsic turbidity of scaffolds formed by natural biomaterials such as collagen fibers prevents high-resolution light microscopy in depth. In this research, we have developed a new method of using light microscopy for penetrative three-dimensional (3-D) visualization of scaffolds formed by collagen, chitosan, or cellulose. First, we applied an optical-clearing solution, FocusClear, to permeate and reduce the turbidity of the scaffolds. The improved photon penetration allowed fluorophores for efficient excitation and emission in the FocusClear solution. Confocal microscopy was applied to achieve cellular-level resolution up to 350 microm for both the fibroblast/collagen and the osteoblast/chitosan constructs and micrometer-level resolution up to 40 microm for the cellulose membrane. The depth of imaging of the cellulose membrane was further improved to 80 microm using two-photon microscopy. Significantly, these voxel-based confocal/two-photon micrographs allowed postrecording image processing via Amira projection algorithms for 3-D visualization and analysis of the scanned region. Although this optical method remains limited in viewing block scaffolds in thin sections, our approach provides a noninvasive way to microscopica...Continue Reading
Confocal fluorescence spectroscopy of subcutaneous cartilage expressing green fluorescent protein versus cutaneous collagen autofluorescence
Molecular interactions of exogenous chemical agents with collagen--implications for tissue optical clearing
Use of glycerol as an optical clearing agent for enhancing photonic transference and detection of Salmonella typhimurium through porcine skin
Composite chondroitin-6-sulfate/dermatan sulfate/chitosan scaffolds for cartilage tissue engineering
Influence of porosity and fibre diameter on the degradation of chitosan fibre-mesh scaffolds and cell adhesion
Functional imaging in bulk tissue specimens using optical emission tomography: fluorescence preservation during optical clearing
Specific requirement of NMDA receptors for long-term memory consolidation in Drosophila ellipsoid body.
Optical clearing of human skin: comparative study of permeability and dehydration of intact and photothermally perforated skin
Penetration kinetics of dimethyl sulphoxide and glycerol in dynamic optical clearing of porcine skin tissue in vitro studied by Fourier transform infrared spectroscopic imaging
Quantitative second harmonic generation imaging and modeling of the optical clearing mechanism in striated muscle and tendon
Differential permeability rate and percent clearing of glucose in different regions in rabbit sclera
Optical clearing of unsectioned specimens for three-dimensional imaging via optical transmission and emission tomography.
3-D visualization and quantitation of microvessels in transparent human colorectal carcinoma [corrected
Three-dimensional optical method for integrated visualization of mouse islet microstructure and vascular network with subcellular-level resolution
ISDoT: in situ decellularization of tissues for high-resolution imaging and proteomic analysis of native extracellular matrix
Vascular labeling of luminescent gold nanorods enables 3-D microscopy of mouse intestinal capillaries
Glycerol-mediated nanostructure modification leading to improved transparency of porous polymeric scaffolds for high performance 3D cell imaging
Cell Imaging in CNS
Here is the latest research on cell imaging and imaging modalities, including light-sheet microscopy, in the central nervous system.