SOLEIL: Single-objective lens inclined light sheet localization microscopy

Shih Te Hung, Jelmer Cnossen, Daniel Fan, Marijn Siemons, Daphne Jurriens, Kristin Grusmayer, Oleg Soloviev, Lukas C. Kapitein, Carlas S. Smith*

*Corresponding author for this work

Research output: Contribution to journalArticleScientificpeer-review

8 Citations (Scopus)
130 Downloads (Pure)

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.

Original languageEnglish
Pages (from-to)3275-3294
JournalBiomedical Optics Express
Volume13
Issue number6
DOIs
Publication statusPublished - 2022

Fingerprint

Dive into the research topics of 'SOLEIL: Single-objective lens inclined light sheet localization microscopy'. Together they form a unique fingerprint.

Cite this