Phase contrast tomography of the mouse cochlea at microfocus x-ray sources

Bartels, Matthias; Hernández, Victor H.; Krenkel, Martin; Moser, Tobias; Salditt, Tim

doi:10.1063/1.4818737

Cited by 64 publications

(75 citation statements)

References 34 publications

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“…35, Figure 4C, Supplemental Figure 8, and Supplemental Video 1). Resolution and contrast sufficed for tracing the basilar membrane ( Figure 4C, inset).…”

Section: Resultsmentioning

confidence: 99%

Optogenetic stimulation of the auditory pathway

et al. 2014

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Auditory prostheses can partially restore speech comprehension when hearing fails. Sound coding with current prostheses is based on electrical stimulation of auditory neurons and has limited frequency resolution due to broad current spread within the cochlea. In contrast, optical stimulation can be spatially confined, which may improve frequency resolution. Here, we used animal models to characterize optogenetic stimulation, which is the optical stimulation of neurons genetically engineered to express the light-gated ion channel channelrhodopsin-2 (ChR2).

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“…35, Figure 4C, Supplemental Figure 8, and Supplemental Video 1). Resolution and contrast sufficed for tracing the basilar membrane ( Figure 4C, inset).…”

Section: Resultsmentioning

confidence: 99%

Optogenetic stimulation of the auditory pathway

et al. 2014

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“…Similar to synchrotron radiation, where the signal‐to‐noise is high enough to detect anatomical and histological details in unstained specimens, and similar to previous studies in other biological samples (Bartels et al., 2013; Krenkel et al., 2015; Töpperwien, Krenkel, Quade, et al., 2016), suitable drying procedures may thus be sufficient to yield sufficient image contrast also for insects and arthropods.…”

Section: Perspectives and Final Considerationsmentioning

confidence: 99%

“…In addition, we experimentally study a range of staining and fixation approaches useful for future studies and provide an outlook into questions that might be answered using micro‐CT in the future. Finally, we propose to especially exploit phase contrast, which has now become a reality also with advanced laboratory μ‐CT (Bartels, Hernandez, Krenkel, Moser, & Salditt, 2013; Töpperwien, Krenkel, Quade, & Salditt, 2016; Töpperwien et al., 2017)…”

Section: Introductionmentioning

confidence: 99%

X‐ray computed tomography and its potential in ecological research: A review of studies and optimization of specimen preparation

Gutiérrez

Ott

Töpperwien

et al. 2018

Ecology and Evolution

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Imaging techniques are a cornerstone of contemporary biology. Over the last decades, advances in microscale imaging techniques have allowed fascinating new insights into cell and tissue morphology and internal anatomy of organisms across kingdoms. However, most studies so far provided snapshots of given reference taxa, describing organs and tissues under “idealized” conditions. Surprisingly, there is an almost complete lack of studies investigating how an organism′s internal morphology changes in response to environmental drivers. Consequently, ecology as a scientific discipline has so far almost neglected the possibilities arising from modern microscale imaging techniques. Here, we provide an overview of recent developments of X‐ray computed tomography as an affordable, simple method of high spatial resolution, allowing insights into three‐dimensional anatomy both in vivo and ex vivo. We review ecological studies using this technique to investigate the three‐dimensional internal structure of organisms. In addition, we provide practical comparisons between different preparation techniques for maximum contrast and tissue differentiation. In particular, we consider the novel modality of phase contrast by self‐interference of the X‐ray wave behind an object (i.e., phase contrast by free space propagation). Using the cricket Acheta domesticus (L.) as model organism, we found that the combination of FAE fixative and iodine staining provided the best results across different tissues. The drying technique also affected contrast and prevented artifacts in specific cases. Overall, we found that for the interests of ecological studies, X‐ray computed tomography is useful when the tissue or structure of interest has sufficient contrast that allows for an automatic or semiautomatic segmentation. In particular, we show that reconstruction schemes which exploit phase contrast can yield enhanced image quality. Combined with suitable specimen preparation and automated analysis, X‐ray CT can therefore become a promising quantitative 3D imaging technique to study organisms′ responses to environmental drivers, in both ecology and evolution.

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“…20 In the first case the resolution is limited by the source spot dimensions whereas in the last case the detector PSF is the limiting factor. As the minimal spot size of the source lies in the range of 4 µm we chose the second approach in order to obtain the highest possible resolution.…”

Section: Laboratory Setupmentioning

confidence: 99%

Phase-contrast tomography of neuronal tissues: from laboratory- to high resolution synchrotron CT

et al. 2016

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Assessing the three-dimensional architecture of neuronal tissues with sub-cellular resolution presents a significant analytical challenge. Overcoming the limitations associated with serial slicing, phase-contrast x-ray tomography has the potential to contribute to this goal. Even compact laboratory CT at an optimized liquid-metal jet microfocus source combined with suitable phase-retrieval algorithms and preparation protocols can yield renderings with single cell sensitivity in millimeter sized brain areas of mouse. Here, we show the capabilities of the setup by imaging a Golgi-Cox impregnated mouse brain. Towards higher resolution we extend these studies at our recently upgraded waveguide-based cone-beam holo-tomography instrument GINIX at DESY. This setup allows high resolution recordings with adjustable field of view and resolution, down to the voxel sizes in the range of a few ten nanometers. The recent results make us confident that important issues of neuronal connectivity can be addressed by these methods, and that 3D (virtual) histology with nanoscale resolution will become an attractive modality for neuroscience research.

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Phase contrast tomography of the mouse cochlea at microfocus x-ray sources

Cited by 64 publications

References 34 publications

Optogenetic stimulation of the auditory pathway

Optogenetic stimulation of the auditory pathway

X‐ray computed tomography and its potential in ecological research: A review of studies and optimization of specimen preparation

Phase-contrast tomography of neuronal tissues: from laboratory- to high resolution synchrotron CT

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