Determination of blood cell subtype concentrations from frozen whole blood samples using TruCount beads

Langenskiöld, Cecilia; Mellgren, Karin; Abrahamsson, Jonas; Bemark, Mats

doi:10.1002/cyto.b.21390

Cited by 17 publications

(22 citation statements)

References 16 publications

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“…Cryopreservation also provides further benefit in that biospecimens can be analyzed in large batches, minimizing overall analytical variability; this is especially important for studies in which collection occurs over a long period of time, or a large number of samples are obtained. Recently, we and others showed that cryopreserved peripheral blood, a biospecimen that is simple and inexpensive to prepare, is comparable to fresh blood for the enumeration of a number of leukocyte types, including monocytes, neutrophils, B‐lymphocytes, T‐cells, natural killer, and (NK)‐cells; our study also showed that these measures were also comparable to that obtained from cryopreserved peripheral blood mononuclear cells (PBMCs) or a hematology analyzer. However, many of these cell types, especially monocytes, T‐cells, and NK‐cells, are known to represent distinct groups of subsets that have specific roles in maintaining health and preventing disease.…”

supporting

confidence: 72%

Cryopreserved whole blood for the quantification of monocyte, T‐cell and NK‐cell subsets, and monocyte receptor expression by multi‐color flow cytometry: A methodological study based on participants from the canadian longitudinal study on aging

Kohli

2018

Cytometry Pt A

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Immunophenotyping by multi-color flow cytometry is arguably the best tool to identify and quantify distinct cell lineages from the peripheral blood and other biological fluids/tissues. Effective in both clinical and research settings, it can be used to estimate the frequency of a given cell type or measure its phenotypic or functional properties. Normally, immunophenotyping is performed in fresh or fractionated blood (i.e., PBMCs) the same day, or within 24 hours of collection; however, this may not be feasible for all study designs. We have previously shown that cryopreserved blood, a biospecimen that is simple and inexpensive to prepare, is comparable to fresh blood for the enumeration of major leukocyte cell types. For the following study, we sought to extend these observations to distinct subsets of: monocytes (classical, intermediate, and non-classical), T-cells (CD4/CD8 naïve, central and effector memory, senescent, and terminally differentiated, and regulatory T-cells), and NK-cells (CD56 bright and dim); we also examined the expression of monocyte cell-surface receptors CX3CR1, CCR2, TLR2, and TLR4. Our results indicate that cryopreserved blood is comparable to fresh blood; with exception to relatively rare subsets and lowly expressed receptors, the absolute or relative frequency of cell subsets generally correlated >0.80 between blood types, while monocyte receptor expressed was mostly >0.70. Furthermore, the day-to-day coefficient of variation for most cell subsets and parameters was below 20%. Given these findings, we suggest that cryopreserved peripheral blood be given greater consideration for studies in which the quantification of distinct leukocyte subsets is required. © 2018 International Society for Advancement of Cytometry.

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supporting

confidence: 72%

Cryopreserved whole blood for the quantification of monocyte, T‐cell and NK‐cell subsets, and monocyte receptor expression by multi‐color flow cytometry: A methodological study based on participants from the canadian longitudinal study on aging

Kohli

2018

Cytometry Pt A

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“…Of the two previous studies (16,17) that also compared absolute counts between fresh and cryopreserved blood, one also found these estimates to be significantly lower in cryopreserved blood, albeit to a lesser extent than our findings (17). We observed a significant discrepancy between absolute counts from fresh and cryopreserved blood, which was far less dramatic for estimates relative to CD45 and almost non-existent for estimates relative to PBMCs.…”

Section: Cytometry Part B: Clinical Cytometrycontrasting

confidence: 59%

“…Based on previous reports (16,21), we suspected that the cell washing step employed following staining, fixation and lysis might be the culprit. Based on previous reports (16,21), we suspected that the cell washing step employed following staining, fixation and lysis might be the culprit.…”

Section: Washing Following Staining Fixation and Lysis Of Cryopresermentioning

confidence: 96%

“…The validity of cryopreserved blood for use with multi-colour flow cytometry has been published previously, but with exception to a recent study by Langenski€ old and colleagues (16), these studies either focused on the functional capacity of immune cell following cryopreservation (13)(14)(15) or were limited to frequency estimates for only a small number of subsets, most commonly lymphocytes (17,18). The validity of cryopreserved blood for use with multi-colour flow cytometry has been published previously, but with exception to a recent study by Langenski€ old and colleagues (16), these studies either focused on the functional capacity of immune cell following cryopreservation (13)(14)(15) or were limited to frequency estimates for only a small number of subsets, most commonly lymphocytes (17,18).…”

Section: Cytometry Part B: Clinical Cytometrymentioning

confidence: 99%

“…Tested for use with functional assays (13)(14)(15) and the enumeration of leukocytes subsets (16)(17)(18), cryopreserved blood appears to be a suitable biospecimen for downstream experiments involving flow cytometry. Tested for use with functional assays (13)(14)(15) and the enumeration of leukocytes subsets (16)(17)(18), cryopreserved blood appears to be a suitable biospecimen for downstream experiments involving flow cytometry.…”

mentioning

confidence: 99%

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Essential Fluidics for a Flow Cytometer

Suthanthiraraj,

Shreve,

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Flow cytometry is an inherently fluidic process that flows particles on a one‐by‐one basis through a sensing region to discretely measure their optical and physical properties. It can be used to analyze particles ranging in size from nanoparticles to whole organisms (e.g., zebrafish). It has particular value for blood analysis, and thus most instruments are fluidically optimized for particles that are comparable in size to a typical blood cell. The principles of fluid dynamics allow for particles of such size to be precisely positioned in flow as they pass through sensing regions that are tens of microns in length at linear velocities of meters per second. Such fluidic systems enable discrete analysis of cell‐sized particles at rates approaching 100 kHz. For larger particles, the principles of fluidics greatly reduce the achievable rates, but such high rates of data acquisition for cell‐sized particles allow rapid collection of information on many thousands to millions of cells and provides for research and clinical measurements of both rare and common cell populations with a high degree of statistical confidence. Additionally, flow cytometers can accurately count particles via the use of volumetric sample delivery and can be coupled with high‐throughput sampling technologies to greatly increase the rate at which independent samples can be delivered to the system. Due to the combination of high analysis rates, sensitive multiparameter measurements, high‐throughput sampling, and accurate counting, flow cytometry analysis is the gold standard for many critical applications in clinical, research, pharmaceutical, and environmental areas. Beyond the power of flow cytometry as an analytical technique, the fluidic pathway can be coupled with a sorting mechanism to collect particles based on desired properties. We present an overview of fluidic systems that enable flow cytometry–based analysis and sorting. We introduce historical approaches, explanations of commonly implemented fluidics, and brief discussions of potential future fluidics where appropriate. © 2024 Wiley Periodicals LLC.

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Determination of blood cell subtype concentrations from frozen whole blood samples using TruCount beads

Cited by 17 publications

References 16 publications

Cryopreserved whole blood for the quantification of monocyte, T‐cell and NK‐cell subsets, and monocyte receptor expression by multi‐color flow cytometry: A methodological study based on participants from the canadian longitudinal study on aging

Cryopreserved whole blood for the quantification of monocyte, T‐cell and NK‐cell subsets, and monocyte receptor expression by multi‐color flow cytometry: A methodological study based on participants from the canadian longitudinal study on aging

A comprehensive assessment of immunophenotyping performed in cryopreserved peripheral whole blood

Essential Fluidics for a Flow Cytometer

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