SOLEIL
Single-objective lens inclined light sheet localization microscopy
Shih Te Hung (TU Delft - Team Carlas Smith)
JP Cnossen (TU Delft - Team Carlas Smith)
DANIEL FAN (TU Delft - Team Carlas Smith)
Marijn Siemons (Universiteit Utrecht)
Daphne Jurriens (Universiteit Utrecht)
K.S. Grussmayer (TU Delft - BN/Kristin Grussmayer Lab)
Oleg Soloviev (TU Delft - Team Michel Verhaegen, Flexible Optical B.V.)
Lukas C. Kapitein (Universiteit Utrecht)
C.S. Smith (TU Delft - Team Carlas Smith, ImPhys/Computational Imaging)
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Abstract
High-NA light sheet illumination can improve the resolution of single-molecule localization microscopy (SMLM) by reducing the background fluorescence. These approaches currently require custom-made sample holders or additional specialized objectives, which makes the sample mounting or the optical system complex and therefore reduces the usability of these approaches. Here, we developed a single-objective lens-inclined light sheet microscope (SOLEIL) that is capable of 2D and 3D SMLM in thick samples. SOLEIL combines oblique illumination with point spread function PSF engineering to enable dSTORM imaging in a wide variety of samples. SOLEIL is compatible with standard sample holders and off-the-shelve optics and standard high NA objectives. To accomplish optimal optical sectioning we show that there is an ideal oblique angle and sheet thickness. Furthermore, to show what optical sectioning delivers for SMLM we benchmark SOLEIL against widefield and HILO microscopy with several biological samples. SOLEIL delivers in 15 μm thick Caco2-BBE cells a 374% higher intensity to background ratio and a 54% improvement in the estimated CRLB compared to widefield illumination, and a 184% higher intensity to background ratio and a 20% improvement in the estimated CRLB compared to HILO illumination.