scholarly journals A simple, inexpensive and multi-scale 3-D fluorescent test sample for optical sectioning microscopies

2021 ◽  
Author(s):  
Ilya Olevsko ◽  
Kaitlin Szederkenyi ◽  
Jennifer Corridon ◽  
Aaron Au ◽  
Brigitte Delhomme ◽  
...  
Keyword(s):  
Quantitative Imaging ◽  
Three Dimensional ◽  
Emission Spectra ◽  
Light Sheet ◽  
Test Sample ◽  
Detection Sensitivity ◽  
List Type ◽  
Optical Sectioning ◽  
Tissue Paper ◽  
Core Facilities

ABSTRACTFluorescence standards allow for quality control and for the comparison of data sets across instruments and laboratories in applications of quantitative fluorescence. For example, users of microscopy core facilities expect a homogenous and time-invariant illumination and a uniform detection sensitivity, which are prerequisites for quantitative imaging analysis, particle tracking or fluorometric pH or Ca2+-concentration measurements. Similarly, confirming the three-dimensional (3-D) resolution of optical sectioning micro-scopes prior to volumetric reconstructions calls for a regular calibration with a standardised point source. Typically, the test samples required for such calibration measurements are different ones, and they depend much on the very microscope technique used. Also, the ever-increasing choice among these techniques increases the demand for comparison and metrology across instruments. Here, we advocate and demonstrate the multiple uses of a surprisingly versatile and simple 3-D test sample that can complement existing and much more expensive calibration samples: simple commercial tissue paper labelled with a fluorescent highlighter pen. We provide relevant sample characteristics and show examples ranging from the sub-µm to cm scale, acquired on epifluorescence, confocal, image scanning, two-photon (2P) and light-sheet microscopes.Graphical abstractPyranine-labeled tissue paper, imaged upon 405-nm epifluorescence excitation through a 455LP LP dichroic and 465LP emission filter. Objective ×20/NA0.25. Overlaid are the normalised absorbance (dashed) and emission spectra (through line), respectively. In the present work we show that this “primitive” and inexpensive three-dimensional (3-D) test sample is a surprisingly versatile and powerful tool for quality assessment, comparison across microscopes as well as routine metrology for optical sectioning techniques, both for research labs and imaging core facilities.Research highlights-highlighter-pen marked tissue paper is a surprisingly powerful and versatile test sample for 3-D fluorescence microscopies-standard tissue paper presents features ranging from 400 nm to centimetres-our sample can simultaneously be used for testing intensity, field homogeneity, resolution, optical sectioning and image contrast-it is easy to prepare, versatile, photostable and inexpensive

scholarly journals Three-Dimensional Microscopy by Milling with Ultraviolet Excitation

2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Jiaming Guo ◽  
Camille Artur ◽  
Jason L. Eriksen ◽  
David Mayerich
Keyword(s):  
Three Dimensional ◽  
Light Sheet ◽  
Clinical Settings ◽  
Three Dimensional Imaging ◽  
Image Stack ◽  
Light Sheet Microscopy ◽  
Ultraviolet Excitation ◽  
Tissue Changes ◽  
Block Face ◽  
Core Facilities

Abstract Analysis of three-dimensional biological samples is critical to understanding tissue function and the mechanisms of disease. Many chronic conditions, like neurodegenerative diseases and cancers, correlate with complex tissue changes that are difficult to explore using two-dimensional histology. While three-dimensional techniques such as confocal and light-sheet microscopy are well-established, they are time consuming, require expensive instrumentation, and are limited to small tissue volumes. Three-dimensional microscopy is therefore impractical in clinical settings and often limited to core facilities at major research institutions. There would be a tremendous benefit to providing clinicians and researchers with the ability to routinely image large three-dimensional tissue volumes at cellular resolution. In this paper, we propose an imaging methodology that enables fast and inexpensive three-dimensional imaging that can be readily integrated into current histology pipelines. This method relies on block-face imaging of paraffin-embedded samples using deep-ultraviolet excitation. The imaged surface is then ablated to reveal the next tissue section for imaging. The final image stack is then aligned and reconstructed to provide tissue models that exceed the depth and resolution achievable with modern three-dimensional imaging systems.

scholarly journals A noninvasive fluorescence imaging-based platform measures 3D anisotropic extracellular diffusion

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Peng Chen ◽  
Xun Chen ◽  
R. Glenn Hepfer ◽  
Brooke J. Damon ◽  
Changcheng Shi ◽  
...  
Keyword(s):  
Anisotropic Diffusion ◽  
Biological Systems ◽  
Three Dimensional ◽  
Molecular Transport ◽  
Light Sheet ◽  
Fundamental Limitations ◽  
Disease Mechanisms ◽  
Dimensional Diffusion ◽  
Compositional Changes ◽  
Diffusion Tensors

AbstractDiffusion is a major molecular transport mechanism in biological systems. Quantifying direction-dependent (i.e., anisotropic) diffusion is vitally important to depicting how the three-dimensional (3D) tissue structure and composition affect the biochemical environment, and thus define tissue functions. However, a tool for noninvasively measuring the 3D anisotropic extracellular diffusion of biorelevant molecules is not yet available. Here, we present light-sheet imaging-based Fourier transform fluorescence recovery after photobleaching (LiFT-FRAP), which noninvasively determines 3D diffusion tensors of various biomolecules with diffusivities up to 51 µm2 s−1, reaching the physiological diffusivity range in most biological systems. Using cornea as an example, LiFT-FRAP reveals fundamental limitations of current invasive two-dimensional diffusion measurements, which have drawn controversial conclusions on extracellular diffusion in healthy and clinically treated tissues. Moreover, LiFT-FRAP demonstrates that tissue structural or compositional changes caused by diseases or scaffold fabrication yield direction-dependent diffusion changes. These results demonstrate LiFT-FRAP as a powerful platform technology for studying disease mechanisms, advancing clinical outcomes, and improving tissue engineering.

scholarly journals A Review on Biosensors and Recent Development of Nanostructured Materials-Enabled Biosensors

Sensors ◽  
2021 ◽  
Vol 21 (4) ◽  
pp. 1109
Author(s):  
Varnakavi. Naresh ◽  
Nohyun Lee
Keyword(s):  
High Surface ◽  
Three Dimensional ◽  
Biological Response ◽  
Volume Ratio ◽  
Recent Decade ◽  
Disease Diagnosis ◽  
Electrical Signal ◽  
Detection Sensitivity ◽  
Wide Range ◽  
Color Tunability

A biosensor is an integrated receptor-transducer device, which can convert a biological response into an electrical signal. The design and development of biosensors have taken a center stage for researchers or scientists in the recent decade owing to the wide range of biosensor applications, such as health care and disease diagnosis, environmental monitoring, water and food quality monitoring, and drug delivery. The main challenges involved in the biosensor progress are (i) the efficient capturing of biorecognition signals and the transformation of these signals into electrochemical, electrical, optical, gravimetric, or acoustic signals (transduction process), (ii) enhancing transducer performance i.e., increasing sensitivity, shorter response time, reproducibility, and low detection limits even to detect individual molecules, and (iii) miniaturization of the biosensing devices using micro-and nano-fabrication technologies. Those challenges can be met through the integration of sensing technology with nanomaterials, which range from zero- to three-dimensional, possessing a high surface-to-volume ratio, good conductivities, shock-bearing abilities, and color tunability. Nanomaterials (NMs) employed in the fabrication and nanobiosensors include nanoparticles (NPs) (high stability and high carrier capacity), nanowires (NWs) and nanorods (NRs) (capable of high detection sensitivity), carbon nanotubes (CNTs) (large surface area, high electrical and thermal conductivity), and quantum dots (QDs) (color tunability). Furthermore, these nanomaterials can themselves act as transduction elements. This review summarizes the evolution of biosensors, the types of biosensors based on their receptors, transducers, and modern approaches employed in biosensors using nanomaterials such as NPs (e.g., noble metal NPs and metal oxide NPs), NWs, NRs, CNTs, QDs, and dendrimers and their recent advancement in biosensing technology with the expansion of nanotechnology.

scholarly journals Three-dimensional quantification of twisting in the Arabidopsis petiole

2021 ◽  
Author(s):  
Yuta Otsuka ◽  
Hirokazu Tsukaya
Keyword(s):  
Cross Sections ◽  
Three Dimensional ◽  
Light Sheet ◽  
Underlying Mechanism ◽  
Two Dimensions ◽  
Observation Angle ◽  
Differential Growth ◽  
Light Sheet Microscopy ◽  
Definition Of ◽  
Twisting Angle

AbstractOrganisms have a variety of three-dimensional (3D) structures that change over time. These changes include twisting, which is 3D deformation that cannot happen in two dimensions. Twisting is linked to important adaptive functions of organs, such as adjusting the orientation of leaves and flowers in plants to align with environmental stimuli (e.g. light, gravity). Despite its importance, the underlying mechanism for twisting remains to be determined, partly because there is no rigorous method for quantifying the twisting of plant organs. Conventional studies have relied on approximate measurements of the twisting angle in 2D, with arbitrary choices of observation angle. Here, we present the first rigorous quantification of the 3D twisting angles of Arabidopsis petioles based on light sheet microscopy. Mathematical separation of bending and twisting with strict definition of petiole cross-sections were implemented; differences in the spatial distribution of bending and twisting were detected via the quantification of angles along the petiole. Based on the measured values, we discuss that minute degrees of differential growth can result in pronounced twisting in petioles.

scholarly journals Apolar and polar transitions drive the conversion between amoeboid and mesenchymal shapes in melanoma cells

2015 ◽  
Vol 26 (22) ◽  
pp. 4163-4170 ◽  
Author(s):  
Sam Cooper ◽  
Amine Sadok ◽  
Vicky Bousgouni ◽  
Chris Bakal
Keyword(s):  
Quantitative Imaging ◽  
Three Dimensional ◽  
Shape Space ◽  
Melanoma Cells ◽  
Collagen Matrices ◽  
Morphological Heterogeneity ◽  
Rho Family Gtpases ◽  
Metastatic Process ◽  
Rho Family ◽  
Single Living

Melanoma cells can adopt two functionally distinct forms, amoeboid and mesenchymal, which facilitates their ability to invade and colonize diverse environments during the metastatic process. Using quantitative imaging of single living tumor cells invading three-dimensional collagen matrices, in tandem with unsupervised computational analysis, we found that melanoma cells can switch between amoeboid and mesenchymal forms via two different routes in shape space—an apolar and polar route. We show that whereas particular Rho-family GTPases are required for the morphogenesis of amoeboid and mesenchymal forms, others are required for transitions via the apolar or polar route and not amoeboid or mesenchymal morphogenesis per se. Altering the transition rates between particular routes by depleting Rho-family GTPases can change the morphological heterogeneity of cell populations. The apolar and polar routes may have evolved in order to facilitate conversion between amoeboid and mesenchymal forms, as cells are either searching for, or attracted to, particular migratory cues, respectively.

scholarly journals Using Stage- and Slit-Scanning to Improve Contrast and Optical Sectioning in Dual-View Inverted Light Sheet Microscopy (diSPIM)

2016 ◽  
Vol 231 (1) ◽  
pp. 26-39 ◽  
Author(s):  
Abhishek Kumar ◽  
Ryan Christensen ◽  
Min Guo ◽  
Panos Chandris ◽  
William Duncan ◽  
...  
Keyword(s):  
Light Sheet ◽  
Optical Sectioning ◽  
Light Sheet Microscopy ◽  
Slit Scanning

scholarly journals First Asian population study of stereotactic body radiation therapy for ventricular arrhythmias

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Li-Ting Ho ◽  
Jenny Ling-Yu Chen ◽  
Hsing-Min Chan ◽  
Yu-Cheng Huang ◽  
Mao-Yuan Su ◽  
...  
Keyword(s):  
Radiation Therapy ◽  
Three Dimensional ◽  
Asian Population ◽  
Target Volume ◽  
Dual Energy ◽  
Detection Sensitivity ◽  
Dual Energy Ct ◽  
Stereotactic Body Radiation ◽  
Integrated Boost ◽  
Monitoring Treatment

AbstractWe report the first Asian series on stereotactic body radiation (SBRT) for refractory ventricular arrhythmia (VA) in Taiwanese patients. Three-dimensional electroanatomic maps, delayed-enhancement magnetic resonance imaging (DE-MRI), and dual-energy computed tomography (CT) were used to identify scar substrates. The main target volume was treated with a single radiation dose of 25 Gy and the margin volume received 20 Gy using simultaneous integrated boost delivered by the Varian TrueBeam system. Efficacy was assessed according to VA events recorded by an implantable cardioverter-defibrillator (ICD) or a 24-h Holter recorder. Pre- and post-radiation therapy imaging studies were performed. From February 2019 to December 2019, seven patients (six men, one woman; mean age, 55 years) were enrolled and treated. One patient died of hepatic failure. In the remaining six patients, at a median follow-up of 14.5 months, the VA burden and ICD shocks significantly decreased (only one patient with one ICD shock after treatment). Increased intensity on DE-MRI might be associated with a lower risk for VA recurrence, whereas dual-energy CT had lower detection sensitivity. No acute or minimal late adverse events occurred. In patients with refractory VA, SBRT is associated with a marked reduction in VA burden and ICD shocks, and DE-MRI might be useful for monitoring treatment effects.

Cubic spline-based depth-dependent localization of mitochondria–endoplasmic reticulum contacts by three-dimensional light-sheet super-resolution microscopy

The Analyst ◽  
2021 ◽  
Author(s):  
Yucheng Sun ◽  
Seungah Lee ◽  
Seong Ho Kang
Keyword(s):  
Endoplasmic Reticulum ◽  
Calcium Homeostasis ◽  
Three Dimensional ◽  
Super Resolution ◽  
Light Sheet ◽  
Cubic Spline ◽  
Contact Distance ◽  
Super Resolution Microscopy

The contact distance between mitochondria (Mito) and endoplasmic reticulum (ER) has received considerable attention owing to their crucial function in maintaining lipid and calcium homeostasis. Herein, cubic spline algorithm-based depth-dependent...

scholarly journals Three-Dimensional Cross-Sectional Light-Sheet Microscopy Imaging of the Inflamed Mouse Gut

2017 ◽  
Vol 153 (4) ◽  
pp. 898-900 ◽  
Author(s):  
Sebastian Zundler ◽  
Anika Klingberg ◽  
Daniela Schillinger ◽  
Sarah Fischer ◽  
Clemens Neufert ◽  
...  
Keyword(s):  
Three Dimensional ◽  
Light Sheet ◽  
Cross Sectional ◽  
Microscopy Imaging ◽  
Light Sheet Microscopy
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