Soleil: single-objective lens inclined light sheet localization microscopy

Shih-Te Hung; Jelmer Cnossen; Daniel Fan; Marijn Siemons; Daphne Jurriens; Kristin Grußmayer; Oleg Soloviev; Lukas C Kapitein; Carlas S Smith

doi:10.1364/BOE.451634

Soleil: single-objective lens inclined light sheet localization microscopy

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

Delft University of Technology

Research output: Contribution to journal › Article › Academic › peer-review

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 language	English
Pages (from-to)	3275-3294
Number of pages	20
Journal	Biomedical Optics Express
Volume	13
Issue number	6
DOIs	https://doi.org/10.1364/BOE.451634
Publication status	Published - 1 Jun 2022

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10.1364/BOE.451634Licence: Other

boe-13-6-3275Final published version, 34 MBLicence: Other

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title = "Soleil: single-objective lens inclined light sheet localization microscopy",

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.",

author = "Shih-Te Hung and Jelmer Cnossen and Daniel Fan and Marijn Siemons and Daphne Jurriens and Kristin Gru{\ss}mayer and Oleg Soloviev and Kapitein, {Lukas C} and Smith, {Carlas S}",

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T2 - single-objective lens inclined light sheet localization microscopy

AU - Hung, Shih-Te

AU - Cnossen, Jelmer

AU - Fan, Daniel

AU - Siemons, Marijn

AU - Jurriens, Daphne

AU - Grußmayer, Kristin

AU - Soloviev, Oleg

AU - Kapitein, Lukas C

AU - Smith, Carlas S

PY - 2022/6/1

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N2 - 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.

AB - 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.

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EP - 3294

JO - Biomedical Optics Express

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Soleil: single-objective lens inclined light sheet localization microscopy

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