This mode captures super-resolution 2D images at high speed with incredible contrast. The TIRF-SIM mode enables Total Internal Reflection Fluorescence observation at double the resolution of 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
The 3D-SIM mode generates structured illumination patterns in three dimensions to deliver a two-fold improvement in lateral and axial resolutions. Two reconstruction methods (“slice” and “stack”) are available to optimize results according to application requirements (e.g. sample thickness, speed, etc.). Slice reconstruction is suitable for imaging living cells at specific depths, as it allows axial super-resolution imaging with optical sectioning at 300 nm resolution. Stack reconstruction, based on Gustafsson’s theory, is suitable for acquisition of volume data as it 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 super-resolution microscope enables 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 indirectly immunolabeled for keratin intermediate filaments and visualized with Alexa Fluor® 488 conjugated secondary antibodies.
Reconstruction method: Stack
Photo courtesy of: Dr. Reinhard Windoffer, RWTH Aachen University