Methodological challenges of optical tweezers-based X-ray fluorescence imaging of biological model organisms at synchrotron facilities

2015 ◽  
Vol 22 (4) ◽  
pp. 1096-1105 ◽  
Author(s):  
Eva Vergucht ◽  
Toon Brans ◽  
Filip Beunis ◽  
Jan Garrevoet ◽  
Stephen Bauters ◽  
...  
Keyword(s):  
Synchrotron Radiation ◽  
Optical Tweezers ◽  
Single Cells ◽  
Biological Model ◽  
Model Organisms ◽  
X Ray ◽  
Research Fields ◽  
Highly Sensitive ◽  
Resolution Level

Recently, a radically new synchrotron radiation-based elemental imaging approach for the analysis of biological model organisms and single cells in their naturalin vivostate was introduced. The methodology combines optical tweezers (OT) technology for non-contact laser-based sample manipulation with synchrotron radiation confocal X-ray fluorescence (XRF) microimaging for the first time at ESRF-ID13. The optical manipulation possibilities and limitations of biological model organisms, the OT setup developments for XRF imaging and the confocal XRF-related challenges are reported. In general, the applicability of the OT-based setup is extended with the aim of introducing the OT XRF methodology in all research fields where highly sensitivein vivomulti-elemental analysis is of relevance at the (sub)micrometre spatial resolution level.

scholarly journals In vivo X-ray elemental imaging of single cell model organisms manipulated by laser-based optical tweezers

2015 ◽  
Vol 5 (1) ◽  
Author(s):  
Eva Vergucht ◽  
Toon Brans ◽  
Filip Beunis ◽  
Jan Garrevoet ◽  
Maarten De Rijcke ◽  
...  
Keyword(s):  
Single Cell ◽  
Optical Tweezers ◽  
Cell Model ◽  
Model Organisms ◽  
X Ray ◽  
Elemental Imaging ◽  
Single Cell Model

F-59 X-ray Imaging on Biological Model Organisms Using Micro and Nano X-ray Fluorescence

2009 ◽  
Vol 24 (2) ◽  
pp. 167-167
Author(s):  
B. De Samber ◽  
K. De Schamphelaere ◽  
B. Masschaele ◽  
S. Vanblaere ◽  
F. Vanhaecke ◽  
...  
Keyword(s):  
Biological Model ◽  
Model Organisms ◽  
X Ray ◽  
X Ray Imaging

scholarly journals Construction of 3D Cellular Composites with Stem Cells Derived from Adipose Tissue and Endothelial Cells by Use of Optical Tweezers in a Natural Polymer Solution

Materials ◽  
2019 ◽  
Vol 12 (11) ◽  
pp. 1759 ◽  
Author(s):  
Takehiro Yamazaki ◽  
Toshifumi Kishimoto ◽  
Paweł Leszczyński ◽  
Koichiro Sadakane ◽  
Takahiro Kenmotsu ◽  
...  
Keyword(s):  
Stem Cells ◽  
Endothelial Cells ◽  
Adipose Tissue ◽  
Optical Tweezers ◽  
Single Cells ◽  
Three Dimensional ◽  
Cell Types ◽  
Cellular Interactions ◽  
Research Fields ◽  
Wide Range

To better understand the regulation and function of cellular interactions, three-dimensional (3D) assemblies of single cells and subsequent functional analysis are gaining popularity in many research fields. While we have developed strategies to build stable cellular structures using optical tweezers in a minimally invasive state, methods for manipulating a wide range of cell types have yet to be established. To mimic organ-like structures, the construction of 3D cellular assemblies with variety of cell types is essential. Our recent studies have shown that the presence of nonspecific soluble polymers in aqueous solution is the key to creating stable 3D cellular assemblies efficiently. The present study further expands on the construction of 3D single cell assemblies using two different cell types. We have successfully generated 3D cellular assemblies, using GFP-labeled adipose tissue-derived stem cells and endothelial cells by using optical tweezers. Our findings will support the development of future applications to further characterize cellular interactions in tissue regeneration.

Strategy of an in vivo Study of the Evolution of Drugs via X-Ray Diffraction with Synchrotron Radiation

2018 ◽  
Vol 82 (7) ◽  
pp. 895-897
Author(s):  
M. R. Sharafutdinov ◽  
B. P. Tolochko ◽  
K. E. Kuper ◽  
A. I. Ancharov
Keyword(s):  
Synchrotron Radiation ◽  
In Vivo Study ◽  
X Ray Diffraction ◽  
X Ray

New methods for semiconductor charge-diffusion-length measurements using synchrotron radiation

1998 ◽  
Vol 5 (3) ◽  
pp. 874-876 ◽  
Author(s):  
J. Kohagura ◽  
T. Cho ◽  
M. Hirata ◽  
T. Okamura ◽  
T. Tamano ◽  
...  
Keyword(s):  
Synchrotron Radiation ◽  
Three Dimensional ◽  
Semiconductor Detectors ◽  
Fusion Plasma ◽  
X Ray ◽  
New Methods ◽  
Research Fields ◽  
Diffusion Effects ◽  
Position Sensitive ◽  
Radiation Research

The extension of a new theory on the X-ray energy response of semiconductor detectors is carried out to characterize the X-ray response of a multichannel semiconductor detector fabricated on one silicon wafer. Recently, these multichannel detectors have been widely utilized for position-sensitive observations in various research fields, including synchrotron radiation research and fusion-plasma investigations. This article represents the verification of the physics essentials of a proposed theory on the X-ray response of semiconductor detectors. The three-dimensional charge-diffusion effects on the adjoining detector-channel signals are experimentally demonstrated at the Photon Factory for two types of multichannel detectors. These findings are conveniently applicable for measuring diffusion lengths for industrial requirements.

In vivo microscopic x-ray imaging in rat and mouse using synchrotron radiation

2008 ◽  
Author(s):  
Keiji Umetani ◽  
Takashi Sakurai ◽  
Takeshi Kondoh
Keyword(s):  
Synchrotron Radiation ◽  
X Ray ◽  
X Ray Imaging

scholarly journals An event-based paradigm for analyzing fluorescent astrocyte activity uncovers novel single-cell and population-level physiology

2018 ◽  
Author(s):  
Yizhi Wang ◽  
Nicole V. DelRosso ◽  
Trisha Vaidyanathan ◽  
Michael Reitman ◽  
Michelle K. Cahill ◽  
...  
Keyword(s):  
Ex Vivo ◽  
Single Cells ◽  
Region Of Interest ◽  
Population Level ◽  
Analytical Framework ◽  
Model Organisms ◽  
Imaging Data ◽  
Fluorescent Indicators ◽  
Event Based

AbstractRecent work examining astrocytic physiology centers on fluorescence imaging approaches, due to development of sensitive fluorescent indicators and observation of spatiotemporally complex calcium and glutamate activity. However, the field remains hindered in fully characterizing these dynamics, both within single cells and at the population-level, because of the insufficiency of current region-of-interest-based approaches to describe activity that is often spatially unfixed, size-varying, and propagative. Here, we present a paradigm-shifting analytical framework that releases astrocyte biologists from ROI-based tools. Astrocyte Quantitative Analysis (AQuA) software enables users to take an event-based approach to accurately capture and quantify the irregular activity observed in astrocyte imaging datasets. We apply AQuA to a range of ex vivo and in vivo imaging data, and uncover previously undescribed physiological phenomena in each. Since AQuA is data-driven and based on machine learning principles, it can be applied across model organisms, fluorescent indicators, experimental modes, and imaging resolutions and speeds, enabling researchers to elucidate fundamental astrocyte physiology.

scholarly journals In vivo super-resolution RESOLFT microscopy of Drosophila melanogaster

eLife ◽  
2016 ◽  
Vol 5 ◽  
Author(s):  
Sebastian Schnorrenberg ◽  
Tim Grotjohann ◽  
Gerd Vorbrüggen ◽  
Alf Herzig ◽  
Stefan W Hell ◽  
...  
Keyword(s):  
Drosophila Melanogaster ◽  
Fluorescent Protein ◽  
Single Cells ◽  
Super Resolution ◽  
Time Lapse ◽  
Live Cell ◽  
Model Organisms ◽  
Alpha Tubulin ◽  
Light Levels

Despite remarkable developments in diffraction unlimited super-resolution microscopy, in vivo nanoscopy of tissues and model organisms is still not satisfactorily established and rarely realized. RESOLFT nanoscopy is particularly suited for live cell imaging because it requires relatively low light levels to overcome the diffraction barrier. Previously, we introduced the reversibly switchable fluorescent protein rsEGFP2, which facilitated fast RESOLFT nanoscopy (<xref ref-type="bibr" rid="bib10">Grotjohann et al., 2012</xref>). In that study, as in most other nanoscopy studies, only cultivated single cells were analyzed. Here, we report on the use of rsEGFP2 for live-cell RESOLFT nanoscopy of sub-cellular structures of intact Drosophila melanogaster larvae and of resected tissues. We generated flies expressing fusion proteins of alpha-tubulin and rsEGFP2 highlighting the microtubule cytoskeleton in all cells. By focusing through the intact larval cuticle, we achieved lateral resolution of <60 nm. RESOLFT nanoscopy enabled time-lapse recordings comprising 40 images and facilitated recordings 40 µm deep within fly tissues.

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