scholarly journals Optical Projection Tomography Using a Commercial Microfluidic System

Micromachines ◽  
2020 ◽  
Vol 11 (3) ◽  
pp. 293
Author(s):  
Wenhao Du ◽  
Cheng Fei ◽  
Junliang Liu ◽  
Yongfu Li ◽  
Zhaojun Liu ◽  
...  
Keyword(s):  
Three Dimensional ◽  
Numerical Aperture ◽  
Microfluidic System ◽  
Depth Of Field ◽  
Objective Lens ◽  
Flow Mode ◽  
Wide Field ◽  
Optical Projection Tomography ◽  
Microfluidic Systems ◽  
Optical Projection

Optical projection tomography (OPT) is the direct optical equivalent of X-ray computed tomography (CT). To obtain a larger depth of field, traditional OPT usually decreases the numerical aperture (NA) of the objective lens to decrease the resolution of the image. So, there is a trade-off between sample size and resolution. Commercial microfluidic systems can observe a sample in flow mode. In this paper, an OPT instrument is constructed to observe samples. The OPT instrument is combined with commercial microfluidic systems to obtain a three-dimensional and time (3D + T)/four-dimensional (4D) video of the sample. “Focal plane scanning” is also used to increase the images’ depth of field. A series of two-dimensional (2D) images in different focal planes was observed and compared with images simulated using our program. Our work dynamically monitors 3D OPT images. Commercial microfluidic systems simulate blood flow, which has potential application in blood monitoring and intelligent drug delivery platforms. We design an OPT adaptor to perform OPT on a commercial wide-field inverted microscope (Olympusix81). Images in different focal planes are observed and analyzed. Using a commercial microfluidic system, a video is also acquired to record motion pictures of samples at different flow rates. To our knowledge, this is the first time an OPT setup has been combined with a microfluidic system.

scholarly journals Optical projection tomography as a quantitative tool for analysis of cell morphology and density in 3D hydrogels

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Birhanu Belay ◽  
Janne T. Koivisto ◽  
Jenny Parraga ◽  
Olli Koskela ◽  
Toni Montonen ◽  
...  
Keyword(s):  
Cell Cultures ◽  
Cell Morphology ◽  
Cell Biology ◽  
Three Dimensional ◽  
Cellular Responses ◽  
Living Cells ◽  
Gellan Gum ◽  
Wide Field ◽  
Optical Projection Tomography ◽  
Optical Projection

AbstractAssessing cell morphology and function, as well as biomaterial performance in cell cultures, is one of the key challenges in cell biology and tissue engineering (TE) research. In TE, there is an urgent need for methods to image actual three-dimensional (3D) cell cultures and access the living cells. This is difficult using established optical microscopy techniques such as wide-field or confocal microscopy. To address the problem, we have developed a new protocol using Optical Projection Tomography (OPT) to extract quantitative and qualitative measurements from hydrogel cell cultures. Using our tools, we demonstrated the method by analyzing cell response in three different hydrogel formulations in 3D with 1.5 mm diameter samples of: gellan gum (GG), gelatin functionalized gellan gum (gelatin-GG), and Geltrex. We investigated cell morphology, density, distribution, and viability in 3D living cells. Our results showed the usability of the method to quantify the cellular responses to biomaterial environment. We observed that an elongated morphology of cells, thus good material response, in gelatin-GG and Geltrex hydrogels compared with basic GG. Our results show that OPT has a sensitivity to assess in real 3D cultures the differences of cellular responses to the properties of biomaterials supporting the cells.

High-resolution 3-D and 4-D imaging using wide-field CCD-based microscopy

1993 ◽  
Vol 51 ◽  
pp. 150-151
Author(s):  
Jason R. Swedlow ◽  
Bethe A. Scalettar ◽  
John W. Sedat ◽  
David A. Agard
Keyword(s):  
Fluorescent Proteins ◽  
Three Dimensional ◽  
Numerical Aperture ◽  
High Sensitivity ◽  
Objective Lens ◽  
Data Sets ◽  
Wide Field ◽  
Data Set ◽  
Image Stack ◽  
A Charge

Because of the limited numerical aperture of a light microscope objective lens, every image recorded from a microscope is blurred and therefore degraded. This problem is particularly acute when a full three-dimensional image stack is viewed in projection. The blurred image is a convolution between every light source in an object and the point-spread function (PSF) of the objective lens and can be mathematically calculated or empirically measured. To eliminate blurring, we record three- and four-dimensional images with a charge-coupled device (CCD) -based computerized optical sectioning microscope and mathematically deconvolve out-of-focus photons using the appropriate three-dimensional, empirically measured PSF. The use of the empirical PSF is important because of the presence of a partial confocal effect in a wide field microscope caused by the presence of the field diaphragm. The advantage in this method is that collection and restoration of out-of-focus photons results in high sensitivity and resolution.In order to follow nuclear and chromosome dynamics, we have injected Drosophila melanogaster embryos with various fluorescent proteins and then monitored the distribution of these proteins during mitosis by recording a series of three-dimensional data sets at regular time intervals-- a four dimensional data set.

scholarly journals Recurrent neural network-based volumetric fluorescence microscopy

2021 ◽  
Vol 10 (1) ◽  
Author(s):  
Luzhe Huang ◽  
Hanlong Chen ◽  
Yilin Luo ◽  
Yair Rivenson ◽  
Aydogan Ozcan
Keyword(s):  
Neural Network ◽  
Image Reconstruction ◽  
Fluorescence Microscopy ◽  
Recurrent Network ◽  
Sample Volume ◽  
Depth Of Field ◽  
Objective Lens ◽  
Wide Field ◽  
Volumetric Imaging ◽  
3D Image Reconstruction

AbstractVolumetric imaging of samples using fluorescence microscopy plays an important role in various fields including physical, medical and life sciences. Here we report a deep learning-based volumetric image inference framework that uses 2D images that are sparsely captured by a standard wide-field fluorescence microscope at arbitrary axial positions within the sample volume. Through a recurrent convolutional neural network, which we term as Recurrent-MZ, 2D fluorescence information from a few axial planes within the sample is explicitly incorporated to digitally reconstruct the sample volume over an extended depth-of-field. Using experiments on C. elegans and nanobead samples, Recurrent-MZ is demonstrated to significantly increase the depth-of-field of a 63×/1.4NA objective lens, also providing a 30-fold reduction in the number of axial scans required to image the same sample volume. We further illustrated the generalization of this recurrent network for 3D imaging by showing its resilience to varying imaging conditions, including e.g., different sequences of input images, covering various axial permutations and unknown axial positioning errors. We also demonstrated wide-field to confocal cross-modality image transformations using Recurrent-MZ framework and performed 3D image reconstruction of a sample using a few wide-field 2D fluorescence images as input, matching confocal microscopy images of the same sample volume. Recurrent-MZ demonstrates the first application of recurrent neural networks in microscopic image reconstruction and provides a flexible and rapid volumetric imaging framework, overcoming the limitations of current 3D scanning microscopy tools.

scholarly journals Application of microfluidic systems for neural differentiation of cells

2019 ◽  
Vol 2 (4) ◽  
pp. 370-381
Author(s):  
Zahra Hesari ◽  
Fatemeh Mottaghitalab ◽  
Akram Shafiee ◽  
Masoud Soleymani ◽  
Rasoul Dinarvand ◽  
...  
Keyword(s):  
Stem Cells ◽  
Small Molecules ◽  
Neuronal Differentiation ◽  
Neural Differentiation ◽  
Three Dimensional ◽  
Microfluidic System ◽  
Microfluidic Systems ◽  
Cell Culture Systems ◽  
Novel Model ◽  
Dimensional Cell

Neural differentiation of stem cells is an important issue in development of central nervous system. Different methods such as chemical stimulation with small molecules, scaffolds, and microRNA can be used for inducing the differentiation of neural stem cells. However, microfluidic systems with the potential to induce neuronal differentiation have established their reputation in the field of regenerative medicine. Organization of microfluidic system represents a novel model that mimic the physiologic microenvironment of cells among other two and three dimensional cell culture systems. Microfluidic system has patterned and well-organized structure that can be combined with other differentiation techniques to provide optimal conditions for neuronal differentiation of stem cells. In this review, different methods for effective differentiation of stem cells to neuronal cells are summarized. The efficacy of microfluidic systems in promoting neuronal differentiation is also addressed.

scholarly journals Extended Dual-Focus Microscopy for Ratiometric-Based 3D Movement Tracking

2020 ◽  
Vol 10 (18) ◽  
pp. 6243
Author(s):  
Seohyun Lee ◽  
Hyuno Kim ◽  
Hideo Higuchi
Keyword(s):  
Drug Delivery ◽  
High Resolution ◽  
Optical Axis ◽  
Three Dimensional ◽  
Virus Transmission ◽  
Living Cells ◽  
Depth Of Field ◽  
Objective Lens ◽  
Accurate Localization ◽  
Movement Tracking

Imaging the three-dimensional movement of small organelles in living cells can provide key information for the dynamics of drug delivery and virus transmission in biomedical disciplines. To stably monitor such intracellular motion using microscope, long depth of field along optical axis and accurate three-dimensional tracking are simultaneously required. In the present work, we suggest an extended dual-focus optics microscopy system by combining a bifocal plane imaging scheme and objective lens oscillation, which enables accurate localization for a long axial range. The proposed system exploits high-resolution functionality by concatenating partial calibration result acquired each axial imaging level, maintaining the practical advantages of ratiometric method.

Sleeve gastrectomy, but not duodenojejunostomy, preserves total beta-cell mass in Goto-Kakizaki rats evaluated by three-dimensional optical projection tomography

2015 ◽  
Vol 30 (2) ◽  
pp. 532-542 ◽  
Author(s):  
Eivind Grong ◽  
Bård Kulseng ◽  
Ingerid Brænne Arbo ◽  
Christoffer Nord ◽  
Maria Eriksson ◽  
...  
Keyword(s):  
Sleeve Gastrectomy ◽  
Beta Cell ◽  
Cell Mass ◽  
Three Dimensional ◽  
Beta Cell Mass ◽  
Optical Projection Tomography ◽  
Optical Projection ◽  
Total Beta

Label‐free optical projection tomography for quantitative three‐dimensional anatomy of mouse embryo

2019 ◽  
Vol 12 (7) ◽  
Author(s):  
Sungbea Ban ◽  
Nam Hyun Cho ◽  
Eunjung Min ◽  
Jung Kweon Bae ◽  
Yujin Ahn ◽  
...  
Keyword(s):  
Mouse Embryo ◽  
Three Dimensional ◽  
Optical Projection Tomography ◽  
Label Free ◽  
Optical Projection

Dual Modal Three dimensional Imaging of Single Cell using Optical Projection Tomography Microscope

2009 ◽  
Author(s):  
Qin Miao ◽  
Benjamin Hawthorne ◽  
Michael Meyer ◽  
J. Richard Rahn ◽  
Thomas Neumann ◽  
...  
Keyword(s):  
Single Cell ◽  
Three Dimensional ◽  
Optical Projection Tomography ◽  
Three Dimensional Imaging ◽  
Optical Projection

scholarly journals OptiJ: Open-source optical projection tomography of large organ samples

2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Pedro P. Vallejo Ramirez ◽  
Joseph Zammit ◽  
Oliver Vanderpoorten ◽  
Fergus Riche ◽  
Francois-Xavier Blé ◽  
...  
Keyword(s):  
Open Source ◽  
Low Cost ◽  
Three Dimensional ◽  
Optical Projection Tomography ◽  
Optical Components ◽  
Adult Mice ◽  
Optical Projection ◽  
Set Up ◽  
Imagej Plugin ◽  
Lung Lobes

Abstract The three-dimensional imaging of mesoscopic samples with Optical Projection Tomography (OPT) has become a powerful tool for biomedical phenotyping studies. OPT uses visible light to visualize the 3D morphology of large transparent samples. To enable a wider application of OPT, we present OptiJ, a low-cost, fully open-source OPT system capable of imaging large transparent specimens up to 13 mm tall and 8 mm deep with 50 µm resolution. OptiJ is based on off-the-shelf, easy-to-assemble optical components and an ImageJ plugin library for OPT data reconstruction. The software includes novel correction routines for uneven illumination and sample jitter in addition to CPU/GPU accelerated reconstruction for large datasets. We demonstrate the use of OptiJ to image and reconstruct cleared lung lobes from adult mice. We provide a detailed set of instructions to set up and use the OptiJ framework. Our hardware and software design are modular and easy to implement, allowing for further open microscopy developments for imaging large organ samples.

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