Development of Spectral Imaging Cytometry

Vorobjev, Ivan A.; Kussanova, Aigul; Barteneva, Natasha S.

doi:10.1007/978-1-0716-3020-4_1

Cited by 5 publications

(2 citation statements)

References 79 publications

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“…New image-based flow cytometers also come equipped with image-based spectral sorting. Spectral flow cytometry could aid andrologists in deriving more information from a single sample and offers an improved framework for cellular characterization and analysis [ 238 , 239 ], which could enable a new generation of sperm sorting techniques, such as sorting for superior morphology sperm. This could be a bridge to help improve human IVF, which currently highly relies on ICSI as opposed to traditional IVF.…”

Section: The Climax—the Future Is Now!mentioning

confidence: 99%

Biomarker-based human and animal sperm phenotyping: the good, the bad and the ugly

Sutovsky,

Hamilton,

Zigo

et al. 2024

Biology of Reproduction

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Conventional, brightfield-microscopic semen analysis provides important baseline information about sperm quality of an individual; however, it falls short of identifying subtle subcellular and molecular defects in cohorts of “bad”, defective human and animal spermatozoa with seemingly normal phenotypes. To bridge this gap, it is desirable to increase the precision of andrological evaluation in humans and livestock animals by pursuing advanced biomarker-based imaging methods. This review, spiced up with occasional classic movie references but seriously scholastic at the same time, focuses mainly on the biomarkers of altered male germ cell proteostasis resulting in post-testicular carryovers of proteins associated with ubiquitin-proteasome system. Also addressed are sperm redox homeostasis, epididymal sperm maturation, sperm-seminal plasma interactions and sperm surface glycosylation. Zinc ion homeostasis-associated biomarkers and sperm-borne components, including the elements of neurodegenerative pathways such as Huntington’s and Alzheimer’s disease, are discussed. Such spectrum of biomarkers, imaged by highly specific vital fluorescent molecular probes, lectins, and antibodies, reveals both obvious and subtle defects of sperm chromatin, DNA and accessory structures of the sperm head and tail. Introduction of next generation image-based flow cytometry into research and clinical andrology will soon enable the incorporation of machine and deep learning algorithms with the end point of developing simple, label-free methods for clinical diagnostics and high throughput phenotyping of spermatozoa in humans and economically important livestock animals.

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Section: The Climax—the Future Is Now!mentioning

confidence: 99%

Biomarker-based human and animal sperm phenotyping: the good, the bad and the ugly

Sutovsky,

Hamilton,

Zigo

et al. 2024

Biology of Reproduction

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“…These developments include high-dimensional conventional cytometers (e.g., Symphony A5, BD), new-generation mass cytometers (CyTOF), and recently developed spectral flow cytometers (Aurora, Cytek and ID7000, Sony Biotechnology) [10,11]. These advances have been accompanied by the development of new fluorescent dyes and metal-tagged antibodies, which together led to an unprecedented dimensionality in the analysis of cell phenotypes.…”

Section: Introductionmentioning

confidence: 99%

Standardized high‐dimensional spectral cytometry protocol and panels for whole blood immune phenotyping in clinical and translational studies

Dott,

Culina,

Chemali

et al. 2023

Cytometry Pt A

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Flow cytometry is the method of choice for immunophenotyping in the context of clinical, translational, and systems immunology studies. Among the latter, the Milieu Intérieur (MI) project aims at defining the boundaries of a healthy immune response to identify determinants of immune response variation. MI used immunophenotyping of a 1000 healthy donor cohort by flow cytometry as a principal outcome for immune variance at steady state. New generation spectral cytometers now enable high‐dimensional immune cell characterization from small sample volumes. Therefore, for the MI 10‐year follow up study, we have developed two high‐dimensional spectral flow cytometry panels for deep characterization of innate and adaptive whole blood immune cells (35 and 34 fluorescent markers, respectively). We have standardized the protocol for sample handling, staining, acquisition, and data analysis. This approach enables the reproducible quantification of over 182 immune cell phenotypes at a single site. We have applied the protocol to discern minor differences between healthy and patient samples and validated its value for application in immunomonitoring studies. Our protocol is currently used for characterization of the impact of age and environmental factors on peripheral blood immune phenotypes of >400 donors from the initial MI cohort.This article is protected by copyright. All rights reserved.

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High-dimensional spectral cytometry panels for whole blood immune phenotyping

Dott,

Culina,

Chemali

et al. 2023

Preprint

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The need to understand the mechanisms and pathways of immune responses in pathogenic conditions such as cancer and autoimmunity requires awareness of natural immune variability in healthy subjects. To this end, various systems immunology studies have been established. Among them, theMilieu Intérieur(MI) study was established to define the boundaries of a healthy immune response and identify determinants of immune response variation. MI used immunophenotyping of a 1000 healthy donor cohort by flow cytometry as a principal outcome for immune variance at steady state. For the 10-year longitudinal MI study, we have developed two high-dimensional spectral flow cytometry panels that allow deep characterization of innate and adaptive whole blood immune cells (35 and 34 fluorescent markers, respectively) and standardized the protocol for sample handling, staining, acquisition, and data analysis. This permits the reproducible quantification of over 182 immune cell phenotypes through robust immunophenotyping at a single site. This highly standardized protocol was applied to samples from patients with autoimmune/inflammatory diseases. It is currently used for characterization of the impact of age and environmental factors on peripheral blood immune phenotypes of >400 donors from the initial MI cohort.

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Development of Spectral Imaging Cytometry

Cited by 5 publications

References 79 publications

Biomarker-based human and animal sperm phenotyping: the good, the bad and the ugly

Biomarker-based human and animal sperm phenotyping: the good, the bad and the ugly

Standardized high‐dimensional spectral cytometry protocol and panels for whole blood immune phenotyping in clinical and translational studies

High-dimensional spectral cytometry panels for whole blood immune phenotyping

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