Light-sheet microscopy in thick media using scanned Bessel beams and two-photon fluorescence excitation

Fahrbach, Florian O.; Gurchenkov, Vasily; Alessandri, Kévin; Nassoy, Pierre; Rohrbach, Alexander

doi:10.1364/oe.21.013824

Cited by 177 publications

(114 citation statements)

References 19 publications

Supporting

Mentioning

113

Contrasting

Order By: Relevance

“…Finally, alternative illumination schemes other than the Gaussian beams modeled within this study have also been reported in the literature (structured illumination, Bessel and Airy beams, multi-photon, etc.) [49][50][51][52], which may be explored using similar methodologies in future studies.…”

Section: Discussionmentioning

confidence: 99%

Assessing the imaging performance of light sheet microscopies in highly scattering tissues

Glaser

Wang

Liu

2016

Biomed. Opt. Express

View full text Add to dashboard Cite

Light sheet microscopy (LSM) has emerged as an opticalimaging method for high spatiotemporal volumetric imaging of relatively transparent samples. While this capability has allowed the technique to be highly impactful in fields such as developmental biology, applications involving highly scattering thick tissues have been largely unexplored. Herein, we employ Monte Carlo simulations to explore the use of LSM for imaging turbid media. In particular, due to its similarity to dual-axis confocal (DAC) microscopy, we compare LSM performance to pointscanned (PS-DAC) and line-scanned (LS-DAC) dual-axis confocal microscopy techniques that have been previously shown to produce highquality images at round-trip optical lengths of ~9 -10 and ~3 -4 respectively. The results of this study indicate that LSM using widefield collection (WF-LSM) provides comparable performance to LS-DAC in thick tissues, due to the fact that they both utilize an illumination beam focused in one dimension (i.e. a line or sheet). On the other hand, LSM using confocal line detection (CL-LSM) is more analogous to PS-DAC microscopy, in which the illumination beam is focused in two dimensions to a point. The imaging depth of LSM is only slightly inferior to DAC (~2 -3 and ~6 -7 optical lengths for WF-LSM and CL-LSM respectively) due to the use of a lower numerical aperture (NA) illumination beam for extended imaging along the illumination axis. Therefore, we conclude that the ability to image deeply is dictated most by the confocality of the microscope technique. In addition, we find that imaging resolution is mostly dependent on the collection NA, and is relatively invariant to imaging depth in a homogeneous scattering medium. Our results indicate that superficial imaging of highly scattering tissues using light sheet microscopy is possible.

show abstract

Section: Discussionmentioning

confidence: 99%

Assessing the imaging performance of light sheet microscopies in highly scattering tissues

Glaser

Wang

Liu

2016

Biomed. Opt. Express

View full text Add to dashboard Cite

show abstract

“…Thus, quasi-Bessel beams can be generated by applying conical phase shifts to a Gaussian beam, using an axicon or a spatial light modulator. Here, a spatial light modulator is used in a configuration described by Fahrbach [14]. We implement the spatial light modulator in a separate beam path using flip mirrors to allow comparison to the original Gaussian illumination.…”

Section: Bessel Beam Illuminationmentioning

confidence: 99%

Gaussian vs. Bessel light-sheets: performance analysis in live large sample imaging

Reidt

Correia

Donnachie

et al. 2017

Optical Trapping and Optical Micromanipulation XIV

View full text Add to dashboard Cite

Lightsheet fluorescence microscopy (LSFM) has rapidly progressed in the past decade from an emerging technology into an established methodology. This progress has largely been driven by its suitability to developmental biology, where it is able to give excellent spatial-temporal resolution over relatively large fields of view with good contrast and low phototoxicity. In many respects it is superseding confocal microscopy. However, it is no magic bullet and still struggles to image deeply in more highly scattering samples. Many solutions to this challenge have been presented, including, Airy and Bessel illumination, 2-photon operation and deconvolution techniques. In this work, we show a comparison between a simple but effective Gaussian beam illumination and Bessel illumination for imaging in chicken embryos. Whilst Bessel illumination is shown to be of benefit when a greater depth of field is required, it is not possible to see any benefits for imaging into the highly scattering tissue of the chick embryo.

show abstract

“…We have used Gaussian-beam light sheets for our imaging because of its simplicity; further improvement to the sectioning capability of the 2p excitation could be achieved by utilizing Bessel-beam light sheets [18][19][20]. Since the side lobes of Bessel beams do not contribute to useful signal generation in 2p excitation, more average laser power would be required for using Bessel-versus Gaussian-beam 2p-SPIM -care then must be exercised to ensure that the extra laser power does not inflict unacceptable photodamage to the live samples under study.…”

Section: Imaging Setup Characterizationmentioning

confidence: 99%

Dynamic structure and protein expression of the live embryonic heart captured by 2-photon light sheet microscopy and retrospective registration

Trivedi¹,

Truong²,

Trinh³

et al. 2015

Biomed. Opt. Express

View full text Add to dashboard Cite

Abstract:We present an imaging and image reconstruction pipeline that captures the dynamic three-dimensional beating motion of the live embryonic zebrafish heart at subcellular resolution. Live, intact zebrafish embryos were imaged using 2-photon light sheet microscopy, which offers deep and fast imaging at 70 frames per second, and the individual optical sections were assembled into a full 4D reconstruction of the beating heart using an optimized retrospective image registration algorithm. This imaging and reconstruction platform permitted us to visualize protein expression patterns at endogenous concentrations in zebrafish gene trap lines.

show abstract

Light-sheet microscopy in thick media using scanned Bessel beams and two-photon fluorescence excitation

Cited by 177 publications

References 19 publications

Assessing the imaging performance of light sheet microscopies in highly scattering tissues

Assessing the imaging performance of light sheet microscopies in highly scattering tissues

Gaussian vs. Bessel light-sheets: performance analysis in live large sample imaging

Dynamic structure and protein expression of the live embryonic heart captured by 2-photon light sheet microscopy and retrospective registration

Contact Info

Product

Resources

About