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Mönkemöller, Viola: Optical super-resolution microscopy of the structure and dynamics of cellular nanopores. 2016
Inhalt
Introduction
Optical nanoscopy
Microscopy techniques
Fluorescence microscopy
Resolution limit
Confocal laser scanning microscopy
Two-photon excitation microscope
Photon-reassignment microscopy
Optical super-resolution microscopy
Stimulated emission depletion (STED)
Single molecule localization microscopy based techniques
Structured Illumination Microscopy
LSECs - Liver Sinusoidal Endothelial Cells
Function and morphology
Human LSECs in 3D-structured illumination microscopy
Preparation of LSECs from rat
Freezing and thawing of LSECs
Confocal microscopy images of LSECs
LSECs imaged by SMLM compared to 3D-SIM
dSTORM on LSECs
Evaluation of the fenestration diameter in dSTORM vs. 3D-SIM
Acquisition speed
Conclusion
LSECs imaged by SMLM and 3D-SIM on the same setup
Correlative SMLM and 3D-SIM on the same setup
Correlative SMLM and 3D-SIM of LSECs
Rapid multi-color 3D-SIM overviews combined with super-resolution of SMLM
Correlation between the membrane and cytoskeleton in LSECs
Fluorescence stains and their effect on living cells
The plasma membrane
Lectins
Wheat germ agglutinin
Ulex europaeus agglutinin
Intercalating dyes
Fenestration dynamics and structural changes of living LSECs
Fenestration dynamics of living LSECs
Effect of different substances on living rat LSECs
Ethanol and paracetamol treatment
Instant SIM and its extension to two-photon microscopy
LSECs imaged by instant SIM
Further super-resolution projects
Visualizing lipid nanodomains in mammalian cells
Open-source image reconstruction of super-resolution SIM data in ImageJ
Nanoparticles as nonfluorescent analogues of fluorophores
Conclusions and outlook
Preparation and staining protocols
LSECs - Liver Sinusoidal Endothelial Cells
LSECs imaged by SMLM compared to 3D-SIM
LSECs imaged by SMLM and 3D-SIM on the same setup
Fluorescence stains and their effect on live cells
Fenestration dynamics and structural changes of living LSECs
Instant SIM and its extension to two-photon microscopy
Visualizing lipid nanodomains in mammalian cells
Open-source image reconstruction of super-resolution SIM data in ImageJ
Nanoparticles as nonfluorescent analogues of fluorophores
Materials and methods
Materials
dSTORM imaging
3D-SIM imaging
Acknowledgment