Out-of-plane integration of a multimode optical fiber for single particle/cell detection at multiple points on a microfluidic device with applications to particle/cell counting, velocimetry, size discrimination and the analysis of single cell lysate injections

Lab on a Chip ◽  
2017 ◽  
Vol 17 (1) ◽  
pp. 145-155 ◽  
Author(s):  
Jalal Sadeghi ◽  
Damith E. W. Patabadige ◽  
Anne H. Culbertson ◽  
Hamid Latifi ◽  
Christopher T. Culbertson
Keyword(s):  
Optical Fiber ◽  
Single Cell ◽  
Single Particle ◽  
Cell Counting ◽  
Cell Detection ◽  
Size Discrimination ◽  
Multiple Points ◽  
Cell Lysate ◽  
Out Of Plane ◽  
Tunneling Mode

An optical fiber tunneling mode is used to create 2 excitation/detection points from a single fiber.

scholarly journals Improved Fabrication of Polymeric Optical Fiber Tweezers for Single Cell Detection

2018 ◽  
Author(s):  
Sandra M. Rodrigues ◽  
Joana S. Paiva ◽  
Rita S. R. Ribeiro ◽  
Olivier Soppera ◽  
Pedro A. S. Jorge
Keyword(s):  
Optical Fiber ◽  
Single Cell ◽  
Cell Detection ◽  
Single Cell Detection

scholarly journals Flexible Minimally Invasive CARS Measurement Method with Tapered Optical Fiber Probe for Single-Cell Application

2022 ◽  
Author(s):  
Tong Wang ◽  
Junfeng Jiang ◽  
Kun Liu ◽  
Shuang Wang ◽  
Panpan Niu ◽  
...  
Keyword(s):  
Minimally Invasive ◽  
Optical Fiber ◽  
Single Cell ◽  
Measurement Method ◽  
Focal Point ◽  
Minimum Size ◽  
Cell Detection ◽  
Fiber Probe ◽  
Optical Fiber Probe ◽  
Tapered Optical Fiber

Abstract We proposed and demonstrated a flexible, endoscopic, and minimally invasive coherent anti-Raman Stokes scattering (CARS) measurement method for single-cell application, employing a tapered optical fiber probe. A few-mode fiber (FMF), whose generated four-wave mixing band is out of CARS signals, was selected to fabricate tapered optical fiber probes, deliver CARS excitation pulses, and collect CARS signals. The adiabatic tapered fiber probe with a diameter of 11.61 µm can focus CARS excitation lights without mismatch at the focal point. The measurements for proof-of-concept were made with methanol, ethanol, cyclohexane, and acetone injected into simulated cells. The experimental results show that the tapered optical fiber probe can detect carbon-hydrogen (C–H) bond-rich substances and their concentration. To our best knowledge, this optical fiber probe provides the minimum size among probes for detecting CARS signals. These results pave the way for minimally invasive live-cell detection in the future.

scholarly journals Digital Cell Counting Device Integrated with a Single-Cell Array

PLoS ONE ◽  
2014 ◽  
Vol 9 (2) ◽  
pp. e89011 ◽  
Author(s):  
Tatsuya Saeki ◽  
Masahito Hosokawa ◽  
Tae-kyu Lim ◽  
Manabu Harada ◽  
Tadashi Matsunaga ◽  
...  
Keyword(s):  
Single Cell ◽  
Cell Counting ◽  
Cell Array ◽  
Counting Device

scholarly journals Epifluorescence-based three-dimensional traction force microscopy

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Lauren Hazlett ◽  
Alexander K. Landauer ◽  
Mohak Patel ◽  
Hadley A. Witt ◽  
Jin Yang ◽  
...  
Keyword(s):  
Spatial Frequency ◽  
Open Source ◽  
Single Particle ◽  
Three Dimensional ◽  
Traction Force ◽  
High Spatial Frequency ◽  
Epifluorescence Microscopy ◽  
Traction Force Microscopy ◽  
Force Microscopy ◽  
Out Of Plane

Abstract We introduce a novel method to compute three-dimensional (3D) displacements and both in-plane and out-of-plane tractions on nominally planar transparent materials using standard epifluorescence microscopy. Despite the importance of out-of-plane components to fully understanding cell behavior, epifluorescence images are generally not used for 3D traction force microscopy (TFM) experiments due to limitations in spatial resolution and measuring out-of-plane motion. To extend an epifluorescence-based technique to 3D, we employ a topology-based single particle tracking algorithm to reconstruct high spatial-frequency 3D motion fields from densely seeded single-particle layer images. Using an open-source finite element (FE) based solver, we then compute the 3D full-field stress and strain and surface traction fields. We demonstrate this technique by measuring tractions generated by both single human neutrophils and multicellular monolayers of Madin–Darby canine kidney cells, highlighting its acuity in reconstructing both individual and collective cellular tractions. In summary, this represents a new, easily accessible method for calculating fully three-dimensional displacement and 3D surface tractions at high spatial frequency from epifluorescence images. We released and support the complete technique as a free and open-source code package.

scholarly journals ICTD: A semi-supervised cell type identification and deconvolution method for multi-omics data

2018 ◽  
Author(s):  
Wennan Chang ◽  
Changlin Wan ◽  
Xiaoyu Lu ◽  
Szu-wei Tu ◽  
Yifan Sun ◽  
...  
Keyword(s):  
Single Cell ◽  
Cell Types ◽  
Training Data ◽  
Marker Genes ◽  
Cell Detection ◽  
Omics Data ◽  
Deconvolution Method ◽  
Cell Type ◽  
Data Set ◽  
Cell Type Specific

AbstractWe developed a novel deconvolution method, namely Inference of Cell Types and Deconvolution (ICTD) that addresses the fundamental issue of identifiability and robustness in current tissue data deconvolution problem. ICTD provides substantially new capabilities for omics data based characterization of a tissue microenvironment, including (1) maximizing the resolution in identifying resident cell and sub types that truly exists in a tissue, (2) identifying the most reliable marker genes for each cell type, which are tissue and data set specific, (3) handling the stability problem with co-linear cell types, (4) co-deconvoluting with available matched multi-omics data, and (5) inferring functional variations specific to one or several cell types. ICTD is empowered by (i) rigorously derived mathematical conditions of identifiable cell type and cell type specific functions in tissue transcriptomics data and (ii) a semi supervised approach to maximize the knowledge transfer of cell type and functional marker genes identified in single cell or bulk cell data in the analysis of tissue data, and (iii) a novel unsupervised approach to minimize the bias brought by training data. Application of ICTD on real and single cell simulated tissue data validated that the method has consistently good performance for tissue data coming from different species, tissue microenvironments, and experimental platforms. Other than the new capabilities, ICTD outperformed other state-of-the-art devolution methods on prediction accuracy, the resolution of identifiable cell, detection of unknown sub cell types, and assessment of cell type specific functions. The premise of ICTD also lies in characterizing cell-cell interactions and discovering cell types and prognostic markers that are predictive of clinical outcomes.

Methodological Implications on Quantitative Studies of Cytocompatibility in Direct Contact with Bioceramic Surfaces

2011 ◽  
Vol 493-494 ◽  
pp. 325-330 ◽  
Author(s):  
J.A. Cortês ◽  
Elena Mavropoulos ◽  
Moema Hausen ◽  
Alexandre Rossi ◽  
J.M. Granjeiro ◽  
...  
Keyword(s):  
Cell Adhesion ◽  
Crystal Violet ◽  
Fluorescent Dyes ◽  
Ceramic Materials ◽  
Neutral Red ◽  
Cell Counting ◽  
Cell Detection ◽  
Porous Hydroxyapatite ◽  
Detection And Counting ◽  
Direct Cell

Cell adhesion, proliferation and differentiation are important specific parameters to be evaluated on biocompatibility studies of candidate biomaterials for clinical applications. Several different methodologies have been employed to study, both qualitative and quantitatively, the direct interactions of ceramic materials with cultured mammal and human cells. However, while quantitatively evaluating cell density, viability and metabolic responses to test materials, several methodological challenges may arise, either by impairing the use of some widely applied techniques, or by generating false or conflicting results. In this work, we tested the inherent interference of different representative calcium phosphate ceramic surfaces (stoichiometric dense and porous hydroxyapatite (HA) and cation-substituted apatite tablets) on different tests for quantitative evaluation of osteoblast adhesion and metabolism, either based on direct cell counting after trypsinization, colorimetric assays (XTT, Neutral Red and Crystal Violet) and fluorescence microscopy. Cell adhesion estimation after trypsinization was highly dependent on the time of treatment, and the group with the highest level of estimated adhesion was inverted from 5 to 20 minutes of exposition to trypsin. Both dense and porous HA samples presented high levels of background adsorption of the Crystal Violet dye, impairing cell detection. HA surfaces also were able to adsorb high levels of fluorescent dyes (DAPI and phalloidin-TRITC), generating backgrounds which, in the case of porous HA, impaired cell detection and counting by image processing software (Image Pro Plus 6.0). We conclude that the choice for the most suitable method for cell detection and estimation is highly dependent on very specific characteristics of the studied material, and methodological adaptations on well established protocols must always be carefully taken on consideration.

scholarly journals Fluorescence enhanced BA-LIFT for single cell detection and isolation

2020 ◽  
Vol 12 (2) ◽  
pp. 025019 ◽  
Author(s):  
A Marquez ◽  
M Gómez-Fontela ◽  
S Lauzurica ◽  
R Candorcio-Simón ◽  
D Munoz-Martin ◽  
...  
Keyword(s):  
Single Cell ◽  
Cell Detection ◽  
Single Cell Detection
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