This mode captures super-resolution 2D images at high speed with incredible contrast. TIRF-SIM mode takes advantage of Total Internal Reflection Fluorescence observation at double the resolution as compared to conventional TIRF microscopes, facilitating a greater understanding of molecular interactions at the cell surface.
Plasma membrane of B16 melanoma cell labeled with YFP
Objective: CFI Apochromat TIRF 100XC Oil (NA 1.49)
Photographed with the cooperation of: Dr. Yasushi Okada, Laboratory for Cell Polarity Regulation, Quantitative Biology Center, RIKEN
Two reconstruction methods are available. Slice reconstruction allows axial super-resolution imaging with optical sectioning at 300 nm resolution in live-cell specimens; stack reconstruction can image thicker specimens with higher contrast than slice reconstruction.
Bacillus subtilis bacterium stained with membrane dye Nile Red (red), and expressing the cell division protein DivIVA fused to GFP (green).
The superior resolution of N-SIM system allows for accurate localization of the protein during division.
Reconstruction method: Slice
Photos courtesy of: Drs. Henrik Strahl and Leendert Hamoen, Centre for Bacterial Cell Biology, Newcastle University
Mouse keratinocyte labeled with an antibody against keratin intermediate filaments and stained with an Alexa Fluor® 488 conjugated second antibody.
Reconstruction method: Stack
Photos courtesy of: Dr. Reinhard Windoffer, RWTH Aachen University