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Cells
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The Applications of Flow Cytometry: Advances, Challenges, and Trends
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The Applications of Flow Cytometry: Advances, Challenges, and Trends

A special issue of Cells (ISSN 2073-4409). This special issue belongs to the section "Cell Methods".

Deadline for manuscript submissions: closed (31 August 2024) | Viewed by 14054

Special Issue Editor

Dr. David R. Kaplan

E-Mail Website
Guest Editor
Department of Pathology, Case Western Reserve University, 2854 Sedgewick Road, Shaker Heights, OH, USA
Interests: flow cytometry; signal amplification; cell-specific molecular expression levels; bipolar disorder; major depressive disorder; PTSD; multiple sclerosis; acute myocardial infarction
Special Issues, Collections and Topics in MDPI journals

Special Issue Information

Dear Colleagues,

Flow cytometry is a well-established and user-friendly single-cell technology that simultaneously measures multiple analyte expression patterns in individual cells. Since its development in the 1960s, it has enabled important analytical and clinical breakthroughs in immunology, cell biology, oncology, bacteriology, infectious disease, rheumatology, and molecular biology.

With its widespread application, flow cytometry has made remarkable progress. Recently, advancements in the field include technological innovations (i.e., spectral cytometry, mass cytometry, and imaging cytometry) and methodological innovations in acquisition and analysis. These advances have rendered flow cytometry an invaluable tool in studies of the immune system and other areas of cell biology.

This Special Issue will provide insights into the applications of using flow cytometry, covering the latest advances, current challenges, and future trends. It aims to broaden our understanding of basic research flow cytometry findings and potentially lead in translating new applications or new protocols into clinical strategies.

We would highly welcome the submission of original article, review, or communication on flow cytometry in research biology or medical science. Interested authors should consult the instructions using the following link: https://www.mdpi.com/journal/cells/instructions.

Dr. David R. Kaplan
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. Cells is an international peer-reviewed open access semimonthly 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 2700 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

  • flow cytometry
  • single cell
  • cell function
  • T-cell subset

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Published Papers (3 papers)

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28 pages, 7251 KiB  
Article
The Power of Reagent Titration in Flow Cytometry
by Diana L. Bonilla, Alberta Paul, Jesus Gil-Pulido, Lily M. Park and Maria C. Jaimes
Cells 2024, 13(20), 1677; https://doi.org/10.3390/cells13201677 - 11 Oct 2024
Viewed by 2863
Abstract
Flow cytometry facilitates the detection of multiple cell parameters simultaneously with a high level of resolution and throughput, enabling in-depth immunological evaluations. High data resolution in flow cytometry depends on multiple factors, including the concentration of reagents used in the staining protocol, and [...] Read more.
Flow cytometry facilitates the detection of multiple cell parameters simultaneously with a high level of resolution and throughput, enabling in-depth immunological evaluations. High data resolution in flow cytometry depends on multiple factors, including the concentration of reagents used in the staining protocol, and reagent validation and titration should be the first step in any assay optimization. Titration is the process of finding the concentration of the reagent that best resolves a positive signal from the background, with the saturation of all binding sites, and minimal antibody excess. The titration process involves the evaluation of serial reagent dilutions in cells expressing the antigen target for the tested antibody. The concentration of antibody that provides the highest signal to noise ratio is calculated by plotting the percentage of positive cells and the intensity of the fluorescence of the stained cells with respect to the negative events, in a concentration–response curve. The determination of the optimal antibody concentration is necessary to ensure reliable and reproducible results and is required for each sample type, reagent clone and lot, as well as the methods used for cell collection, staining, and storage conditions. If the antibody dilution is too low, the signal will be too weak to be accurately determined, leading to suboptimal data resolution, high variability across measurements, and the underestimation of the frequency of cells expressing a specific marker. The use of excess antibodies could lead to non-specific binding, reagent misuse, and detector overloading with the signal off scale and higher spillover spreading. In this publication, we summarized the titration fundamentals and best practices, and evaluated the impact of using a different instrument, sample, staining, acquisition, and analysis conditions in the selection of the optimal titer and population resolution. Full article
(This article belongs to the Special Issue The Applications of Flow Cytometry: Advances, Challenges, and Trends)
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Review

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34 pages, 2584 KiB  
Review
Advances and Challenges in Sepsis Management: Modern Tools and Future Directions
by Elena Santacroce, Miriam D’Angerio, Alin Liviu Ciobanu, Linda Masini, Domenico Lo Tartaro, Irene Coloretti, Stefano Busani, Ignacio Rubio, Marianna Meschiari, Erica Franceschini, Cristina Mussini, Massimo Girardis, Lara Gibellini, Andrea Cossarizza and Sara De Biasi
Cells 2024, 13(5), 439; https://doi.org/10.3390/cells13050439 - 2 Mar 2024
Cited by 8 | Viewed by 8838
Abstract
Sepsis, a critical condition marked by systemic inflammation, profoundly impacts both innate and adaptive immunity, often resulting in lymphopenia. This immune alteration can spare regulatory T cells (Tregs) but significantly affects other lymphocyte subsets, leading to diminished effector functions, altered cytokine profiles, and [...] Read more.
Sepsis, a critical condition marked by systemic inflammation, profoundly impacts both innate and adaptive immunity, often resulting in lymphopenia. This immune alteration can spare regulatory T cells (Tregs) but significantly affects other lymphocyte subsets, leading to diminished effector functions, altered cytokine profiles, and metabolic changes. The complexity of sepsis stems not only from its pathophysiology but also from the heterogeneity of patient responses, posing significant challenges in developing universally effective therapies. This review emphasizes the importance of phenotyping in sepsis to enhance patient-specific diagnostic and therapeutic strategies. Phenotyping immune cells, which categorizes patients based on clinical and immunological characteristics, is pivotal for tailoring treatment approaches. Flow cytometry emerges as a crucial tool in this endeavor, offering rapid, low cost and detailed analysis of immune cell populations and their functional states. Indeed, this technology facilitates the understanding of immune dysfunctions in sepsis and contributes to the identification of novel biomarkers. Our review underscores the potential of integrating flow cytometry with omics data, machine learning and clinical observations to refine sepsis management, highlighting the shift towards personalized medicine in critical care. This approach could lead to more precise interventions, improving outcomes in this heterogeneously affected patient population. Full article
(This article belongs to the Special Issue The Applications of Flow Cytometry: Advances, Challenges, and Trends)
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Other

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8 pages, 3442 KiB  
Brief Report
Flow Cytometry as the Tool to Define Peripheral Blood Leukocyte Signatures in Acute EBV Infection
by Pragya Singh, Manisha Gadgeel, Batool AlQanber, Ahmad Farooqi and Süreyya Savaşan
Cells 2024, 13(11), 963; https://doi.org/10.3390/cells13110963 - 3 Jun 2024
Viewed by 1058
Abstract
Primary Epstein–Barr virus (EBV) infection which can manifest as infectious mononucleosis (IM) is commonly acquired during childhood. EBV primarily invades B cells leading to a lytic reaction; the control of the infection is handled by natural killer and T cells in immunocompetent individuals. [...] Read more.
Primary Epstein–Barr virus (EBV) infection which can manifest as infectious mononucleosis (IM) is commonly acquired during childhood. EBV primarily invades B cells leading to a lytic reaction; the control of the infection is handled by natural killer and T cells in immunocompetent individuals. The infection has a wide spectrum of clinical findings and can lead to serious complications in patients with certain underlying immunological dysfunctions. We retrospectively investigated peripheral white blood cell populations’ surface marker characteristics in IM using a comprehensive flow cytometry marker panel. Twenty-one cases of IM and seventeen EBV-seropositive cases without IM serving as controls were included. We observed novel alterations in lymphocyte, neutrophil, and monocyte populations. In addition to increased activated cytotoxic T cells and low B cells, we demonstrated high T-large granular lymphocyte (T-LGL) populations in IM cases. Furthermore, despite T cells’ increased HLA-DR expression, another activation marker, CD11b, was lower in T-LGL populations. Monocytes showed increased CD16 expression; CD64 was higher in neutrophils. Our findings point to monocyte and neutrophil activation which may account for acute clinical features and may contribute to the understanding of IM immunobiology. Furthermore, they may serve as a useful tool in investigating inherited and post-transplant conditions characterized by deficiencies in controlling EBV infection. Full article
(This article belongs to the Special Issue The Applications of Flow Cytometry: Advances, Challenges, and Trends)
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