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. Optional 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 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
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