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Micromachines
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Advances in Microfluidic Flow Cytometry
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Advances in Microfluidic Flow Cytometry

A special issue of Micromachines (ISSN 2072-666X). This special issue belongs to the section "B：Biology and Biomedicine".

Deadline for manuscript submissions: closed (30 April 2023) | Viewed by 13076

Special Issue Editor

Dr. Gus A. Wright

E-Mail Website
Guest Editor
Department of Veterinary Pathobiology and TAMU Flow Cytometry Facility, Texas A&M University, College Station, TX 77843, USA
Interests: flow cytometry and applications; imaging flow cytometry; microfluidics; fluorescence imaging; immune modulation of cancer and sepsis; cell migration

Special Issue Information

Dear Colleagues, 

In recent years, there has been an increase in demand for portable, low-cost, and compact microfluidic diagnostic devices for point-of-care testing. Microfluidic flow cytometry combines the microscale, on-chip capabilities of microfluidics with the powerful single-cell diagnostics of flow cytometry. Modern microfluidic cytometers, flow cytometers, and cell sorters allow for on-chip manipulation of fluid flow, cell focusing, and particle detection within a single portable, compact, self-contained device. The development of these on-chip devices provides an opportunity to deliver high-quality diagnostics in a portable and cost-effective manner to low socio-economic areas and developing countries. For the Micromachines Special Issue, Advances in Microfluidic Flow Cytometry, we invite submissions on all aspects of development and applications of microfluidic flow cytometry and microfluidic cytometry. Examples of topics include new technologies and functionalities of microfluidic flow cytometers, microfluidic cell sorters, microfluidic imaging flow cytometers, microfluidic cytometers, microfluidic imaging cytometers, and diagnostic and research applications for microfluidic cytometers and microfluidic flow cytometers. Contributions covering the engineering, design, research applications, clinical applications of microfluidic flow cytometers, microfluidic cytometers, cell sorters, imaging flow cytometers, and imaging cytometers will be considered.

Dr. Gus A. Wright
Guest Editor

Manuscript Submission Information

Manuscripts should be submitted online at www.mdpi.com by registering and logging in to this website. Once you are registered, click here to go to the submission form. Manuscripts can be submitted until the deadline. All submissions that pass pre-check are peer-reviewed. Accepted papers will be published continuously in the journal (as soon as accepted) and will be listed together on the special issue website. Research articles, review articles as well as short communications are invited. For planned papers, a title and short abstract (about 100 words) can be sent to the Editorial Office for announcement on this website.

Submitted manuscripts should not have been published previously, nor be under consideration for publication elsewhere (except conference proceedings papers). All manuscripts are thoroughly refereed through a single-blind peer-review process. A guide for authors and other relevant information for submission of manuscripts is available on the Instructions for Authors page. Micromachines is an international peer-reviewed open access monthly journal published by MDPI.

Please visit the Instructions for Authors page before submitting a manuscript. The Article Processing Charge (APC) for publication in this open access journal is 2600 CHF (Swiss Francs). Submitted papers should be well formatted and use good English. Authors may use MDPI's English editing service prior to publication or during author revisions.

Keywords

  • microfluidic flow cytometry
  • microfluidic cell sorting
  • microfluidic imaging flow cytometry
  • microfluidic cytometry
  • microfluidic imaging cytometry
  • impedance detection
  • dielectrophoretic focusing
  • acoustic focusing
  • inertial focusing
  • single cell analysis
  • high throughput
  • clinical diagnostics
  • lab on a chip

Published Papers (6 papers)

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Research

10 pages, 795 KiB  
Article
Photoacoustic Flow Cytometry Using Functionalized Microspheres for Selective Detection of Bacteria
by Robert H. Edgar, Anie-Pier Samson, Tori Kocsis and John A. Viator
Micromachines 2023, 14(3), 573; https://doi.org/10.3390/mi14030573 - 28 Feb 2023
Cited by 2 | Viewed by 1384
Abstract
Photoacoustic flow cytometry is a method to detect rare analytes in fluids. We developed photoacoustic flow cytometry to detect pathological cells in body fluids, such as circulating tumor cells or bacteria in blood. In order to induce specific optical absorption in bacteria, we [...] Read more.
Photoacoustic flow cytometry is a method to detect rare analytes in fluids. We developed photoacoustic flow cytometry to detect pathological cells in body fluids, such as circulating tumor cells or bacteria in blood. In order to induce specific optical absorption in bacteria, we use modified bacteriophage that precisely target bacterial species or subspecies for rapid identification. In order to reduce detection variability and to halt the lytic lifescycle that results in lysis of the bacteria, we attached dyed latex microspheres to the tail fibers of bacteriophage that retained the bacterial recognition binding sites. We tested these microsphere complexes using Salmonella enterica (Salmonella) and Escherichia coli (E. coli) bacteria and found robust and specific detection of targeted bacteria. In our work we used LT2, a strain of Salmonella, against K12, a strain of E. coli. Using Det7, a bacteriophage that binds to LT2 and not to K12, we detected an average of 109.3±9.0 of LT2 versus 2.0±1.7 of K12 using red microspheres and 86.7±13.2 of LT2 versus 0.3±0.6 of K12 using blue microspheres. These results confirmed our ability to selectively detect bacterial species using photoacoustic flow cytometry. Full article
(This article belongs to the Special Issue Advances in Microfluidic Flow Cytometry)
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17 pages, 7247 KiB  
Article
Prism Design for Spectral Flow Cytometry
by Zixi Chao, Yong Han, Zeheng Jiao, Zheng You and Jingjing Zhao
Micromachines 2023, 14(2), 315; https://doi.org/10.3390/mi14020315 - 26 Jan 2023
Viewed by 1791
Abstract
Flow cytometers are instruments used for the rapid quantitative analysis of cell suspension. Traditional flow cytometry uses multi-channel filters to detect fluorescence, whereas full-spectrum fluorescence based on dispersion detection is a more effective and accurate method. The application of various dispersion schemes in [...] Read more.
Flow cytometers are instruments used for the rapid quantitative analysis of cell suspension. Traditional flow cytometry uses multi-channel filters to detect fluorescence, whereas full-spectrum fluorescence based on dispersion detection is a more effective and accurate method. The application of various dispersion schemes in flow cytometry spectroscopy has been studied. From the perspective of modern detectors and demand for the miniaturization of flow cytometry, prism dispersion exhibits higher and more uniform light energy utilization, meaning that it is a more suitable dispersion method for small flow cytometers, such as microfluidic flow cytometers. Prism dispersion designs include the size, number, and placement of prisms. By deducing the formula of the final position of light passing through the prism and combining it with the formula of transmittance, the design criteria of the top angle and the incident angle of the prism in pursuit of the optimum transmittance and dispersion index can be obtained. Considering the case of multiple prisms, under the premise of pursuing a smaller size, the optimal design criteria for dispersion system composed of multiple prisms can be obtained. The design of prism dispersion fluorescence detection was demonstrated with a microfluidic flow cytometer, and the effectiveness of the design results was verified by microsphere experiments and practical biological experiments. This design criterion developed in this study is generally applicable to spectral flow cytometers. Full article
(This article belongs to the Special Issue Advances in Microfluidic Flow Cytometry)
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18 pages, 5562 KiB  
Article
A Simple Micromilled Microfluidic Impedance Cytometer with Vertical Parallel Electrodes for Cell Viability Analysis
by Jason Eades, Julianne F. Audiffred, Micah Fincher, Jin-Woo Choi, Steven A. Soper and William Todd Monroe
Micromachines 2023, 14(2), 283; https://doi.org/10.3390/mi14020283 - 22 Jan 2023
Viewed by 1951
Abstract
Microfluidic impedance cytometry has been demonstrated as an effective platform for single cell analysis, taking advantage of microfabricated features and dielectric cell sensing methods. In this study, we present a simple microfluidic device to improve the sensitivity, accuracy, and throughput of single suspension [...] Read more.
Microfluidic impedance cytometry has been demonstrated as an effective platform for single cell analysis, taking advantage of microfabricated features and dielectric cell sensing methods. In this study, we present a simple microfluidic device to improve the sensitivity, accuracy, and throughput of single suspension cell viability analysis using vertical sidewall electrodes fabricated by a widely accessible negative manufacturing method. A microchannel milled through a 75 µm platinum wire, which was embedded into poly-methyl-methacrylate (PMMA), created a pair of parallel vertical sidewall platinum electrodes. Jurkat cells were interrogated in a custom low-conductivity buffer (1.2 ± 0.04 mS/cm) to reduce current leakage and increase device sensitivity. Confirmed by live/dead staining and electron microscopy, a single optimum excitation frequency of 2 MHz was identified at which live and dead cells were discriminated based on the disruption in the cell membrane associated with cell death. At this frequency, live cells were found to exhibit changes in the impedance phase with no appreciable change in magnitude, while dead cells displayed the opposite behavior. Correlated with video microscopy, a computational algorithm was created that could identify cell detection events and determine cell viability status by application of a mathematical correlation method. Full article
(This article belongs to the Special Issue Advances in Microfluidic Flow Cytometry)
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10 pages, 12293 KiB  
Article
Upgraded User-Friendly Image-Activated Microfluidic Cell Sorter Using an Optimized and Fast Deep Learning Algorithm
by Keondo Lee, Seong-Eun Kim, Seokho Nam, Junsang Doh and Wan Kyun Chung
Micromachines 2022, 13(12), 2105; https://doi.org/10.3390/mi13122105 - 29 Nov 2022
Cited by 2 | Viewed by 1924
Abstract
Image-based cell sorting is essential in biological and biomedical research. The sorted cells can be used for downstream analysis to expand our knowledge of cell-to-cell differences. We previously demonstrated a user-friendly image-activated microfluidic cell sorting technique using an optimized and fast deep learning [...] Read more.
Image-based cell sorting is essential in biological and biomedical research. The sorted cells can be used for downstream analysis to expand our knowledge of cell-to-cell differences. We previously demonstrated a user-friendly image-activated microfluidic cell sorting technique using an optimized and fast deep learning algorithm. Real-time isolation of cells was carried out using this technique with an inverted microscope. In this study, we devised a recently upgraded sorting system. The cell sorting techniques shown on the microscope were implemented as a real system. Several new features were added to make it easier for the users to conduct the real-time sorting of cells or particles. The newly added features are as follows: (1) a high-resolution linear piezo-stage is used to obtain in-focus images of the fast-flowing cells; (2) an LED strobe light was incorporated to minimize the motion blur of fast-flowing cells; and (3) a vertical syringe pump setup was used to prevent the cell sedimentation. The sorting performance of the upgraded system was demonstrated through the real-time sorting of fluorescent polystyrene beads. The sorter achieved a 99.4% sorting purity for 15 μm and 10 μm beads with an average throughput of 22.1 events per second (eps). Full article
(This article belongs to the Special Issue Advances in Microfluidic Flow Cytometry)
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10 pages, 2085 KiB  
Article
Rapid Bacterial Motility Monitoring Using Inexpensive 3D-Printed OpenFlexure Microscopy Allows Microfluidic Antibiotic Susceptibility Testing
by Tai The Diep, Sarah Helen Needs, Samuel Bizley and Alexander D. Edwards
Micromachines 2022, 13(11), 1974; https://doi.org/10.3390/mi13111974 - 14 Nov 2022
Cited by 5 | Viewed by 2273
Abstract
Antibiotic susceptibility testing is vital to tackle the emergence and spread of antimicrobial resistance. Inexpensive digital CMOS cameras can be converted into portable digital microscopes using 3D printed x-y-z stages. Microscopic examination of bacterial motility can rapidly detect the response of microbes to [...] Read more.
Antibiotic susceptibility testing is vital to tackle the emergence and spread of antimicrobial resistance. Inexpensive digital CMOS cameras can be converted into portable digital microscopes using 3D printed x-y-z stages. Microscopic examination of bacterial motility can rapidly detect the response of microbes to antibiotics to determine susceptibility. Here, we present a new simple microdevice-miniature microscope cell measurement system for multiplexed antibiotic susceptibility testing. The microdevice is made using melt-extruded plastic film strips containing ten parallel 0.2 mm diameter microcapillaries. Two different antibiotics, ceftazidime and gentamicin, were prepared in Mueller-Hinton agar (0.4%) to produce an antibiotic-loaded microdevice for simple sample addition. This combination was selected to closely match current standard methods for both antibiotic susceptibility testing and motility testing. Use of low agar concentration permits observation of motile bacteria responding to antibiotic exposure as they enter capillaries. This device fits onto the OpenFlexure 3D-printed digital microscope using a Raspberry Pi computer and v2 camera, avoiding need for expensive laboratory microscopes. This inexpensive and portable digital microscope platform had sufficient magnification to detect motile bacteria, yet wide enough field of view to monitor bacteria behavior as they entered antibiotic-loaded microcapillaries. The image quality was sufficient to detect how bacterial motility was inhibited by different concentrations of antibiotic. We conclude that a 3D-printed Raspberry Pi-based microscope combined with disposable microfluidic test strips permit rapid, easy-to-use bacterial motility detection, with potential for aiding detection of antibiotic resistance. Full article
(This article belongs to the Special Issue Advances in Microfluidic Flow Cytometry)
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12 pages, 7892 KiB  
Article
Droplet Microfluidic Technology for the Early and Label-Free Isolation of Highly-Glycolytic, Activated T-Cells
by Claudia Zielke, Adriana J. Gutierrez Ramirez, Kelsey Voss, Maya S. Ryan, Azam Gholizadeh, Jeffrey C. Rathmell and Paul Abbyad
Micromachines 2022, 13(9), 1442; https://doi.org/10.3390/mi13091442 - 1 Sep 2022
Viewed by 2549
Abstract
A label-free, fixation-free and passive sorting method is presented to isolate activated T-cells shortly after activation and prior to the display of activation surface markers. It uses a recently developed sorting platform dubbed “Sorting by Interfacial Tension” (SIFT) that sorts droplets based on [...] Read more.
A label-free, fixation-free and passive sorting method is presented to isolate activated T-cells shortly after activation and prior to the display of activation surface markers. It uses a recently developed sorting platform dubbed “Sorting by Interfacial Tension” (SIFT) that sorts droplets based on pH. After polyclonal (anti-CD3/CD28 bead) activation and a brief incubation on chip, droplets containing activated T-cells display a lower pH than those containing naive cells due to increased glycolysis. Under specific surfactant conditions, a change in pH can lead to a concurrent increase in droplet interfacial tension. The isolation of activated T-cells on chip is hence achieved as flattened droplets are displaced as they encounter a micro-fabricated trench oriented diagonally with respect to the direction of flow. This technique leads to an enrichment of activated primary CD4+ T-cells to over 95% from an initial mixed population of naive cells and cells activated for as little as 15 min. Moreover, since the pH change is correlated to successful activation, the technique allows the isolation of T-cells with the earliest activation and highest glycolysis, an important feature for the testing of T-cell activation modulators and to determine regulators and predictors of differentiation outcomes. Full article
(This article belongs to the Special Issue Advances in Microfluidic Flow Cytometry)
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