Miniaturized modules for light sheet microscopy with low chromatic aberration

Bruns, Thomas; Bauer, Michael; Bruns, Sarah; Meyer, Heiko; Kubin, D.; Schneckenburger, Herbert

doi:10.1111/jmi.12439

Cited by 16 publications

(12 citation statements)

References 14 publications

(18 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…This makes the calculation of highresolution SIM images very difficult. However, it appears possible to combine structured illumination and light sheet microscopy [5] (in an up-modulation mode) and to suppress scattered Figure 7. 3T3 fibroblasts from a multilayer incubated with the mitochondrial marker R123 (5 µM for 30 min); wide field image (a) and SIM image (b) in a sample depth ∆z = 10 µm.…”

Section: Discussionmentioning

confidence: 99%

“…This makes the calculation of high-resolution SIM images very difficult. However, it appears possible to combine structured illumination and light sheet microscopy [5] (in an up-modulation mode) and to suppress scattered light outside the light sheet. In this way, imaging in larger depths of the sample may become possible.…”

Section: Discussionmentioning

confidence: 99%

“…It should be emphasized that in comparison to commercial systems, our present SIM setup is not optimized for the highest speed. However, it has the advantage that it uses a conventional inverted microscope and can be easily combined with other microscopy techniques, e.g., light sheet microscopy [5], as well as spectral imaging or fluorescence lifetime imaging microscopy (FLIM).…”

Section: Discussionmentioning

confidence: 99%

“…This favors LSFM for experiments performed over a long time period. Experimental setups for LSFM range from stand-alone microscopes to miniaturized modules, which can be adapted to various kinds of commercial microscopes and combined with further experimental techniques [5]. Only in the last 25 years have methods been described with a resolution below the Abbe criterion.…”

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Increasing Resolution in Live Cell Microscopy by Structured Illumination (SIM)

et al. 2019

View full text Add to dashboard Cite

In the context of various approaches to super-resolution microscopy, structured illumination microscopy (SIM) offers several advantages: it needs rather low light doses (with a low risk of phototoxicity or photobleaching), is comparably fast and flexible concerning the use of microscopes, objective lenses and cameras, and has potential for 3D imaging. This paper describes an experimental setup for SIM with first diffraction orders of a spectral light modulator (SLM) creating an interference pattern in two dimensions. We kept this system rather compact with a comparably large illuminated object field, validated it with nano-beads and applied it further to living cells for imaging the cytoskeleton, mitochondria or cell nuclei with a resolution slightly above 100 nm. Its advantages, challenges and limitations—concerning cameras, acquisition time, depth of imaging, light exposure, and combining it with further super-resolving methods—are discussed.

show abstract

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Increasing Resolution in Live Cell Microscopy by Structured Illumination (SIM)

et al. 2019

View full text Add to dashboard Cite

show abstract

“…Hence, it is more suitable for low resolution imaging. Other designs integrate a sample cuvette in the form of a capillary with side illumination into a stage inset of an inverted microscope [8][9][10][11]. While this approach is compact and low cost, it provides relatively low axial resolution (~5 µm) and/or demands specific tube-mounted samples.…”

Section: Introductionmentioning

confidence: 99%

sideSPIM – selective plane illumination based on a conventional inverted microscope

Hedde¹,

Malacrida²,

Ahrar³

et al. 2017

Biomed. Opt. Express

View full text Add to dashboard Cite

Previously described selective plane illumination microscopy techniques typically offset ease of use and sample handling for maximum imaging performance or . Also, to reduce cost and complexity while maximizing flexibility, it is highly desirable to implement light sheet microscopy such that it can be added to a standard research microscope instead of setting up a dedicated system. We devised a new approach termed sideSPIM that provides uncompromised imaging performance and easy sample handling while, at the same time, offering new applications of plane illumination towards fluidics and high throughput 3D imaging of multiple specimen. Based on an inverted epifluorescence microscope, all of the previous functionality is maintained and modifications to the existing system are kept to a minimum. At the same time, our implementation is able to take full advantage of the speed of the employed sCMOS camera and piezo stage to record data at rates of up to 5 stacks/s. Additionally, sample handling is compatible with established methods and switching magnification to change the field of view from single cells to whole organisms does not require labor intensive adjustments of the system.

show abstract

A minimal-complexity light-sheet microscope maps network activity in 3D neuronal systems

Wysmolek

Kiessler

Salbaum

et al. 2022

Sci Rep

View full text Add to dashboard Cite

In vitro systems mimicking brain regions, brain organoids, are revolutionizing the neuroscience field. However, characterization of their electrical activity has remained a challenge as it requires readout at millisecond timescale in 3D at single-neuron resolution. While custom-built microscopes used with genetically encoded sensors are now opening this door, a full 3D characterization of organoid neural activity has not been performed yet, limited by the combined complexity of the optical and the biological system. Here, we introduce an accessible minimalistic light-sheet microscope to the neuroscience community. Designed as an add-on to a standard inverted microscope it can be assembled within one day. In contrast to existing simplistic setups, our platform is suited to record volumetric calcium traces. We successfully extracted 4D calcium traces at high temporal resolution by using a lightweight piezo stage to allow for 5 Hz volumetric scanning combined with a processing pipeline for true 3D neuronal trace segmentation. As a proof of principle, we created a 3D connectivity map of a stem cell derived neuron spheroid by imaging its activity. Our fast, low complexity setup empowers researchers to study the formation of neuronal networks in vitro for fundamental and neurodegeneration research.

show abstract

Miniaturized modules for light sheet microscopy with low chromatic aberration

Cited by 16 publications

References 14 publications

Increasing Resolution in Live Cell Microscopy by Structured Illumination (SIM)

Increasing Resolution in Live Cell Microscopy by Structured Illumination (SIM)

sideSPIM – selective plane illumination based on a conventional inverted microscope

A minimal-complexity light-sheet microscope maps network activity in 3D neuronal systems

Contact Info

Product

Resources

About