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.
