Super-resolution localization microscopy provides a powerful tool to study biochemical mechanisms at single molecule level. Although the lateral position of the fluorescent dye molecules can be determined routinely with high precision, measurement of other modalities such as 3D and multicolor without the degradation of the original super-resolved image is still in the focus. In this paper a dual-objective multimodal single molecule localization microscopy (SMLM) technique has been developed, optimized and tested. The proposed optical arrangement can be implemented onto a conventional inverted microscope without serious system modification. The performance of the method was tested using fluorescence beads, F-actin filaments and sarcomere structures. It was shown that the proposed imaging method does not degrade the image quality of the original SMLM 2D image but could provide information on the axial position or emission spectra of the dye molecules.
Gajdos et al., Scientific Reports volume 9, Article number: 798 (2019)
OSLO file: mmSTORM-OSLO
> Optical simulation of the Dual Objective setup.
Blender file: mmSTORM_2inch_2
> 3D rendering of the Dual Objective setup.
LabView mmSTORM-ast-lens: GitLab repository GPLv3
> Calculating cylindrical lens position in normal and dual objective setups.
Gallery (images and blender simulation):