superresolution structured illumination microscopy (sr-sim)  (Carl Zeiss)

Journal: Scientific Reports

Article Title: Imaging of angiogenesis of human umbilical vein endothelial cells by uptake of exosomes secreted from hepatocellular carcinoma cells

doi: 10.1038/s41598-018-24563-0

Figure Lengend Snippet: The isolation, characterization, and observation of exosomes secreted from HepG2 cells. ( a ) The exosomes secreted from HepG2 cells (HepG2-exosomes) isolated by the ExoQuick-TC kit in the bottom of the tubes were found as white pellets. Both white arrows show the exosomes. ( b ) An image of transmission electron microscopy of exosomes secreted from HepG2. ( c ) The size distribution, average size and zeta potential of HepG2-exosomes in distilled water. ( d–h ) The expression of marker molecules such as CD63, CD81, NKG2D and HSP70 on the surface of HepG2-exosomes. ( i ) The protein concentration of exosomes secreted from HepG2, as determined by the BCA method. ( j–m ) The exosomes in HepG2 cells were detected by using a FITC-labeled anti-human CD63 mouse monoclonal antibody. The morphology of the cells ( j ), nuclei labeled with Hoechst33342 ( k ), exosomes labeled with FITC-labeled anti-human CD63 mouse monoclonal antibody exist in HepG2 cells ( l ) and the merged images of the nuclei and exosomes ( m ) are shown. White arrows show the exosomes. ( n,o ) Three-dimensional images of nuclei labeled with Hoechst33342 ( n ) and exosomes bound to the FITC-labeled anti-human CD63 mouse monoclonal antibody ( o ) found in HepG2 cells. White arrows show the exosomes. These figures were obtained using superresolution structured illumination microscopy (SR-SIM, Carl Zeiss).

Article Snippet: After washing the samples with the culture medium three times, they were observed by superresolution structured illumination microscopy (SR-SIM (ELYRA), Carl Zeiss Co., Ltd.).

Techniques: Isolation, Transmission Assay, Electron Microscopy, Zeta Potential Analyzer, Expressing, Marker, Protein Concentration, Labeling, Microscopy

Journal: Scientific Reports

Article Title: Imaging of angiogenesis of human umbilical vein endothelial cells by uptake of exosomes secreted from hepatocellular carcinoma cells

doi: 10.1038/s41598-018-24563-0

Figure Lengend Snippet: The isolation, characterization, and observation of exosomes secreted from HepG2 cells. ( a ) The exosomes secreted from HepG2 cells (HepG2-exosomes) isolated by the ExoQuick-TC kit in the bottom of the tubes were found as white pellets. Both white arrows show the exosomes. ( b ) An image of transmission electron microscopy of exosomes secreted from HepG2. ( c ) The size distribution, average size and zeta potential of HepG2-exosomes in distilled water. ( d–h ) The expression of marker molecules such as CD63, CD81, NKG2D and HSP70 on the surface of HepG2-exosomes. ( i ) The protein concentration of exosomes secreted from HepG2, as determined by the BCA method. ( j–m ) The exosomes in HepG2 cells were detected by using a FITC-labeled anti-human CD63 mouse monoclonal antibody. The morphology of the cells ( j ), nuclei labeled with Hoechst33342 ( k ), exosomes labeled with FITC-labeled anti-human CD63 mouse monoclonal antibody exist in HepG2 cells ( l ) and the merged images of the nuclei and exosomes ( m ) are shown. White arrows show the exosomes. ( n,o ) Three-dimensional images of nuclei labeled with Hoechst33342 ( n ) and exosomes bound to the FITC-labeled anti-human CD63 mouse monoclonal antibody ( o ) found in HepG2 cells. White arrows show the exosomes. These figures were obtained using superresolution structured illumination microscopy (SR-SIM, Carl Zeiss).

Article Snippet: These figures were obtained using superresolution structured illumination microscopy (SR-SIM, Carl Zeiss).

Techniques: Isolation, Transmission Assay, Electron Microscopy, Zeta Potential Analyzer, Expressing, Marker, Protein Concentration, Labeling, Microscopy

Journal: Scientific Reports

Article Title: Real-time two- and three-dimensional imaging of monocyte motility and navigation on planar surfaces and in collagen matrices: roles of Rho

doi: 10.1038/srep25016

Figure Lengend Snippet: ( A ) Maximum intensity projection of collagen type I fibers in a loose matrix (0.8 mg/ml collagen), obtained by picrosirius red staining and superresolution structured illumination microscopy, and snapshot (200 × 400 μm) of fluorescently labeled monocytes, in a loose 3D collagen matrix, migrating in a chemotactic fMLP gradient. Scale bar: 10 μm. ( B ) Migration tracks of monocytes (in a loose collagen type I matrix and chemotactic fMLP gradient) in the absence of inhibitors (control) or in the presence of a ROCK inhibitor (Y-27632) or Rho inhibitor (TAT-C3). ( C ) Migration tracks of monocytes (in a loose collagen type I matrix and chemotactic fMLP gradient) in the presence of a nonmuscle myosin II inhibitor (blebbistatin). To circumvent phototoxicity, cells were labeled with a red fluorescent dye and illuminated with green light. ( D ) Summary data. Data are shown as mean ± s.e.m. for tracked cells (25 per experiment) pooled from two to four independent experiments for each group. * p < 0.05; Kruskal-Wallis one way analysis of variance on ranks and Dunn’s method for post-hoc comparisons.

Article Snippet: 3D images were obtained using an ELYRA S.1 superresolution structured illumination microscopy (SR-SIM) system (Zeiss, Germany), which provides resolution beyond the diffraction limit .

Techniques: Staining, Microscopy, Labeling, Migration, Control

Journal: Scientific Reports

Article Title: Real-time two- and three-dimensional imaging of monocyte motility and navigation on planar surfaces and in collagen matrices: roles of Rho

doi: 10.1038/srep25016

Figure Lengend Snippet: ( A ) Maximum intensity projection of collagen type I fibers in a dense matrix (2.4 mg/ml collagen), obtained by picrosirius red staining and superresolution structured illumination microscopy, and snapshot (200 × 400 μm) of fluorescently labeled monocytes, in a dense 3D collagen matrix, migrating in a chemotactic fMLP gradient. Scale bar: 10 μm. ( B ) Migration tracks of monocytes (in a dense collagen type I matrix and chemotactic fMLP gradient) in the absence of inhibitors (control) or in the presence of a ROCK inhibitor (Y-27632) or Rho inhibitor (TAT-C3). ( C ) Migration tracks of monocytes (in a dense collagen type I matrix and chemotactic fMLP gradient) in the presence of a nonmuscle myosin II inhibitor (blebbistatin). To circumvent phototoxicity, cells were labeled with a red fluorescent dye and illuminated with green light. ( D ) Summary data. Data are shown as mean ± s.e.m. for tracked cells (25 per experiment) pooled from two to six independent experiments for each group. * p < 0.05; Kruskal-Wallis one way analysis of variance on ranks and Dunn’s method for post-hoc comparisons.

Article Snippet: 3D images were obtained using an ELYRA S.1 superresolution structured illumination microscopy (SR-SIM) system (Zeiss, Germany), which provides resolution beyond the diffraction limit .

Techniques: Staining, Microscopy, Labeling, Migration, Control