Combining 3D single molecule localization strategies and sprectral demultiplexing for bioimaging (Conference Presentation)

DAISY combined Supercritical Angle Fluorescence (SAF) detection in order to provide an absolute axial reference to a point spread function engineering approach . The dual view optical setup permits to introduce a strong astigmatism and decouple lateral and axial information, thus extending the axial performances without degrading the lateral precision. This technique provides 3D absolute information over a 1-µm capture range above the glass coverslip and an axial localization precision down to 15 nm with minimal loss of lateral resolution and little sensitivity to field aberrations. We will present sequential and simultaneous up to 3 color 3D imaging in cells.