scholarly journals Cellular lensing and near infrared fluorescent nanosensor arrays to enable chemical efflux cytometry

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Soo-Yeon Cho ◽  
Xun Gong ◽  
Volodymyr B. Koman ◽  
Matthias Kuehne ◽  
Sun Jin Moon ◽  
...  
Keyword(s):  
Near Infrared ◽  
Single Cells ◽  
Microfluidic Channel ◽  
Human Monocyte ◽  
Cellular Heterogeneity ◽  
Label Free ◽  
Nanotube Array ◽  
Chemical Cytometry ◽  
Rich Information ◽  
Non Destructive

AbstractNanosensors have proven to be powerful tools to monitor single cells, achieving spatiotemporal precision even at molecular level. However, there has not been way of extending this approach to statistically relevant numbers of living cells. Herein, we design and fabricate nanosensor array in microfluidics that addresses this limitation, creating a Nanosensor Chemical Cytometry (NCC). nIR fluorescent carbon nanotube array is integrated along microfluidic channel through which flowing cells is guided. We can utilize the flowing cell itself as highly informative Gaussian lenses projecting nIR profiles and extract rich information. This unique biophotonic waveguide allows for quantified cross-correlation of biomolecular information with various physical properties and creates label-free chemical cytometer for cellular heterogeneity measurement. As an example, the NCC can profile the immune heterogeneities of human monocyte populations at attomolar sensitivity in completely non-destructive and real-time manner with rate of ~600 cells/hr, highest range demonstrated to date for state-of-the-art chemical cytometry.

scholarly journals Cellular Lensing and Near Infrared Fluorescent Nanosensor Arrays to Enable Chemical Efflux Cytometry

2020 ◽  
Author(s):  
Soo-Yeon Cho ◽  
Xun Gong ◽  
Volodymyr Koman ◽  
Matthias Kuehne ◽  
Sun Jin Moon ◽  
...  
Keyword(s):  
High Throughput ◽  
Single Molecule ◽  
Near Infrared ◽  
Microfluidic Channel ◽  
Human Monocyte ◽  
Cellular Heterogeneity ◽  
Label Free ◽  
Temporal Precision ◽  
Single Molecule Level ◽  
Chemical Cytometry

Abstract Nanosensor have proven to be powerful tools to monitor single biological cells and organisms, achieving spatial and temporal precision even at the single molecule level. However, there has not been a way of extending this approach to statistically relevant numbers of living cells and organisms. Herein, we design and fabricate a high throughput nanosensor array in a microfluidic channel that addresses this limitation, creating a Nanosensor Chemical Cytometry (NCC). An array of nIR fluorescent single walled carbon nanotube (SWNT) nanosensors is integrated along a microfluidic channel through which a population of flowing cells is guided. We show that one can utilize the flowing cell itself as highly informative Gaussian lenses projecting nIR emission profiles and extract rich information on a per cell basis at high throughput. This unique biophotonic waveguide allows for quantified cross-correlation of the biomolecular information with physical properties such as cellular diameter, refractive index (RI), and eccentricity and creates a label-free chemical cytometer for the measurement of cellular heterogeneity with unprecedented precision. As an example, the NCC can profile the immune response heterogeneities of distinct human monocyte populations at attomolar (10-18 moles) sensitivity in a completely non-destructive and real-time manner with a rate of ~100 cells/frame, highest range demonstrated to date for state of the art chemical cytometry. We demonstrate distinct H2O2 efflux heterogeneities between 330 and 624 attomole/cell·min with cell projected areas between 271 and 263 µm2, eccentricity values between 0.405 and 0.363 and RI values between 1.383 and 1.377 for non-activated and activated human monocytes, respectively. Hence, we show that our nanotechnology based biophotonic cytometer has significant potential and versatility to answer important questions and provide new insight in immunology, cell manufacturing and biopharmaceutical research.

scholarly journals Magnetic levitation of single cells

2015 ◽  
Vol 112 (28) ◽  
pp. E3661-E3668 ◽  
Author(s):  
Naside Gozde Durmus ◽  
H. Cumhur Tekin ◽  
Sinan Guven ◽  
Kaushik Sridhar ◽  
Ahu Arslan Yildiz ◽  
...  
Keyword(s):  
Magnetic Levitation ◽  
Single Cells ◽  
Nonsmall Cell Lung Cancer ◽  
Cellular Heterogeneity ◽  
Cell Populations ◽  
Label Free ◽  
Biological Characterization ◽  
Lung Cancer Cell ◽  
Cellular Density ◽  
Cellular Events

Several cellular events cause permanent or transient changes in inherent magnetic and density properties of cells. Characterizing these changes in cell populations is crucial to understand cellular heterogeneity in cancer, immune response, infectious diseases, drug resistance, and evolution. Although magnetic levitation has previously been used for macroscale objects, its use in life sciences has been hindered by the inability to levitate microscale objects and by the toxicity of metal salts previously applied for levitation. Here, we use magnetic levitation principles for biological characterization and monitoring of cells and cellular events. We demonstrate that each cell type (i.e., cancer, blood, bacteria, and yeast) has a characteristic levitation profile, which we distinguish at an unprecedented resolution of 1 × 10−4g⋅mL−1. We have identified unique differences in levitation and density blueprints between breast, esophageal, colorectal, and nonsmall cell lung cancer cell lines, as well as heterogeneity within these seemingly homogenous cell populations. Furthermore, we demonstrate that changes in cellular density and levitation profiles can be monitored in real time at single-cell resolution, allowing quantification of heterogeneous temporal responses of each cell to environmental stressors. These data establish density as a powerful biomarker for investigating living systems and their responses. Thereby, our method enables rapid, density-based imaging and profiling of single cells with intriguing applications, such as label-free identification and monitoring of heterogeneous biological changes under various physiological conditions, including antibiotic or cancer treatment in personalized medicine.

scholarly journals Encapsulated Cell Dynamics in Droplet Microfluidic Devices with Sheath Flow

Micromachines ◽  
2021 ◽  
Vol 12 (7) ◽  
pp. 839
Author(s):  
Peter E. Beshay ◽  
Ali M. Ibrahim ◽  
Stefanie S. Jeffrey ◽  
Roger T. Howe ◽  
Yasser H. Anis
Keyword(s):  
Flow Field ◽  
High Speed ◽  
Single Cells ◽  
Microfluidic Channel ◽  
Internal Flow ◽  
Shear Stresses ◽  
Rotational Flow ◽  
Label Free ◽  
Cell Dynamics ◽  
Sheath Flow

In this paper we study the dynamics of single cells encapsulated in water-in-oil emulsions in a microchannel. The flow field of a microfluidic channel is coupled to the internal flow field of a droplet through viscous traction at the interface, resulting in a rotational flow field inside the droplet. An encapsulated single cell being subjected to this flow field responds by undergoing multiple orbits, spins, and deformations that depend on its physical properties. Monitoring the cell dynamics, using a high-speed camera, can lead to the development of new label-free methods for the detection of rare cells, based on their biomechanical properties. A sheath flow microchannel was proposed to strengthen the rotational flow field inside droplets flowing in Poiseuille flow conditions. A numerical model was developed to investigate the effect of various parameters on the rotational flow field inside a droplet. The multi-phase flow model required the tracking of the fluid–fluid interface, which deforms over time due to the applied shear stresses. Experiments confirmed the significant effect of the sheath flow rate on the cell dynamics, where the speed of cell orbiting was doubled. Doubling the cell speed can double the amount of extracted biomechanical information from the encapsulated cell, while it remains within the field of view of the camera used.

scholarly journals Integrated Field’s metal microelectrodes based microfluidic impedance cytometry for cell-in-droplet quantification

2018 ◽  
Author(s):  
Jatin Panwar ◽  
Rahul Roy
Keyword(s):  
Single Cell ◽  
Single Cell Analysis ◽  
Single Cells ◽  
Microfluidic Channel ◽  
High Sensitivity ◽  
Cell Detection ◽  
Label Free ◽  
Single Cell Detection ◽  
Electrode Fabrication ◽  
Impedance Cytometry

AbstractMicrofluidic impedance cytometry (MIC) provides a non-optical and label-free method for single cell detection and classification in microfluidics. However, the cleanroom intensive infrastructure required for MIC electrode fabrication limits its wide implementation in microfluidic analysis. To bypass the conventional metal (platinum) electrode fabrication protocol, we fabricated coplanar ‘in-contact’ Field’s metal (icFM) microelectrodes in multilayer elastomer devices with a single photolithography step. Our icFM microelectrodes displayed excellent and comparable performance to the platinum electrodes for detection of single erythrocytes with a lock-in amplifier based MIC setup. We further characterized it for water-in-oil droplets generated in a T-junction microfluidic channel and found high sensitivity and long-term operational stability of these electrodes. Finally, to facilitate droplet based single cell analysis, we demonstrate detection and quantification of single cells entrapped in aqueous droplets.

scholarly journals Differential Sorting of Microparticles Using Spiral Microchannels with Elliptic Configurations

Micromachines ◽  
2020 ◽  
Vol 11 (4) ◽  
pp. 412
Author(s):  
Kaan Erdem ◽  
Vahid Ebrahimpour Ahmadi ◽  
Ali Kosar ◽  
Lütfullah Kuddusi
Keyword(s):  
Cell Sorting ◽  
Microfluidic Channel ◽  
Major Axis ◽  
Label Free ◽  
Flow Rates ◽  
Inertial Focusing ◽  
Size Dependent ◽  
Curved Channels ◽  
Aspect Ratios ◽  
Dependent Cell

Label-free, size-dependent cell-sorting applications based on inertial focusing phenomena have attracted much interest during the last decade. The separation capability heavily depends on the precision of microparticle focusing. In this study, five-loop spiral microchannels with a height of 90 µm and a width of 500 µm are introduced. Unlike their original spiral counterparts, these channels have elliptic configurations of varying initial aspect ratios, namely major axis to minor axis ratios of 3:2, 11:9, 9:11, and 2:3. Accordingly, the curvature of these configurations increases in a curvilinear manner through the channel. The effects of the alternating curvature and channel Reynolds number on the focusing of fluorescent microparticles with sizes of 10 and 20 µm in the prepared suspensions were investigated. At volumetric flow rates between 0.5 and 3.5 mL/min (allowing separation), each channel was tested to collect samples at the designated outlets. Then, these samples were analyzed by counting the particles. These curved channels were capable of separating 20 and 10 µm particles with total yields up to approximately 95% and 90%, respectively. The results exhibited that the level of enrichment and the focusing behavior of the proposed configurations are promising compared to the existing microfluidic channel configurations.

scholarly journals Research Note: Non-destructive Detection of Super Grade Chick Embryos or Hatchlings using Near-infrared Spectroscopy

2021 ◽  
pp. 101189
Author(s):  
Alin Khaliduzzaman ◽  
Ayuko Kashimori ◽  
Tetsuhito Suzuki ◽  
Yuichi Ogawa ◽  
Naoshi Kondo
Keyword(s):  
Infrared Spectroscopy ◽  
Near Infrared Spectroscopy ◽  
Near Infrared ◽  
Research Note ◽  
Chick Embryos ◽  
Non Destructive

Ab Initio Two-sites Occupancy and Broadband Near-Infrared Emission of Cr3+ in Li2MgZrO4

2021 ◽  
Author(s):  
Xue Zhou ◽  
Jinmeng Xiang ◽  
Jiming Zheng ◽  
Xiaoqi Zhao ◽  
Hao Suo ◽  
...  
Keyword(s):  
Plant Growth ◽  
Ab Initio ◽  
Light Source ◽  
Near Infrared ◽  
Broad Band ◽  
Infrared Emission ◽  
Night Vision ◽  
Light Emitting ◽  
Non Destructive ◽  
Near Infrared Emission

Near-infrared (NIR) phosphor-converted light-emitting diodes (pc-LEDs) light source have great potential in non-destructive detection, promoting plant growth and night vision applications, while the discovery of a broad-band NIR phosphor still...

scholarly journals 3D phonon microscopy with sub-micron axial-resolution

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Richard J. Smith ◽  
Fernando Pérez-Cota ◽  
Leonel Marques ◽  
Matt Clark
Keyword(s):  
Signal To Noise Ratio ◽  
Single Cells ◽  
Optical Resolution ◽  
Acquisition Time ◽  
Label Free ◽  
Axial Resolution ◽  
Force Microscopy ◽  
Atomic Force ◽  
Accuracy And Precision ◽  
Good Agreement

AbstractBrillouin light scattering (BLS) is an emerging method for cell imaging and characterisation. It allows elasticity-related contrast, optical resolution and label-free operation. Phonon microscopy detects BLS from laser generated coherent phonon fields to offer an attractive route for imaging since, at GHz frequencies, the phonon wavelength is sub-optical. Using phonon fields to image single cells is challenging as the signal to noise ratio and acquisition time are often poor. However, recent advances in the instrumentation have enabled imaging of fixed and living cells. This work presents the first experimental characterisation of phonon-based axial resolution provided by the response to a sharp edge. The obtained axial resolution is up to 10 times higher than that of the optical system used to take the measurements. Validation of the results are obtained with various polymer objects, which are in good agreement with those obtained using atomic force microscopy. Edge localisation, and hence profilometry, of a phantom boundary is measured with accuracy and precision of approximately 60 nm and 100 nm respectively. Finally, 3D imaging of fixed cells in culture medium is demonstrated.

scholarly journals Cryopreservation of human cancers conserves tumour heterogeneity for single-cell multi-omics analysis

2021 ◽  
Vol 13 (1) ◽  
Author(s):  
Sunny Z. Wu ◽  
Daniel L. Roden ◽  
Ghamdan Al-Eryani ◽  
Nenad Bartonicek ◽  
Kate Harvey ◽  
...  
Keyword(s):  
Single Cell ◽  
Rna Sequencing ◽  
High Throughput ◽  
Single Cells ◽  
Cellular Heterogeneity ◽  
Tumour Heterogeneity ◽  
Fresh Tissue ◽  
Human Cancers ◽  
Cryopreserved Cell ◽  
Single Cell Rna Sequencing

Abstract Background High throughput single-cell RNA sequencing (scRNA-Seq) has emerged as a powerful tool for exploring cellular heterogeneity among complex human cancers. scRNA-Seq studies using fresh human surgical tissue are logistically difficult, preclude histopathological triage of samples, and limit the ability to perform batch processing. This hindrance can often introduce technical biases when integrating patient datasets and increase experimental costs. Although tissue preservation methods have been previously explored to address such issues, it is yet to be examined on complex human tissues, such as solid cancers and on high throughput scRNA-Seq platforms. Methods Using the Chromium 10X platform, we sequenced a total of ~ 120,000 cells from fresh and cryopreserved replicates across three primary breast cancers, two primary prostate cancers and a cutaneous melanoma. We performed detailed analyses between cells from each condition to assess the effects of cryopreservation on cellular heterogeneity, cell quality, clustering and the identification of gene ontologies. In addition, we performed single-cell immunophenotyping using CITE-Seq on a single breast cancer sample cryopreserved as solid tissue fragments. Results Tumour heterogeneity identified from fresh tissues was largely conserved in cryopreserved replicates. We show that sequencing of single cells prepared from cryopreserved tissue fragments or from cryopreserved cell suspensions is comparable to sequenced cells prepared from fresh tissue, with cryopreserved cell suspensions displaying higher correlations with fresh tissue in gene expression. We showed that cryopreservation had minimal impacts on the results of downstream analyses such as biological pathway enrichment. For some tumours, cryopreservation modestly increased cell stress signatures compared to freshly analysed tissue. Further, we demonstrate the advantage of cryopreserving whole-cells for detecting cell-surface proteins using CITE-Seq, which is impossible using other preservation methods such as single nuclei-sequencing. Conclusions We show that the viable cryopreservation of human cancers provides high-quality single-cells for multi-omics analysis. Our study guides new experimental designs for tissue biobanking for future clinical single-cell RNA sequencing studies.

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