Dataset Information

Dynamic blood flow phantom for in vivo liquid biopsy standardization.

ABSTRACT: In vivo liquid biopsy, especially using the photoacoustic (PA) method, demonstrated high clinical potential for early diagnosis of deadly diseases such as cancer, infections, and cardiovascular disorders through the detection of rare circulating tumor cells (CTCs), bacteria, and clots in the blood background. However, little progress has been made in terms of standardization of these techniques, which is crucial to validate their high sensitivity, accuracy, and reproducibility. In the present study, we addressed this important demand by introducing a dynamic blood vessel phantom with flowing mimic normal and abnormal cells. The light transparent silica microspheres were used as white blood cells and platelets phantoms, while hollow polymeric capsules, filled with hemoglobin and melanin, reproduced red blood cells and melanoma CTCs, respectively. These phantoms were successfully used for calibration of the PA flow cytometry platform with high-speed signal processing. The results suggest that these dynamic cell flow phantoms with appropriate biochemical, optical, thermal, and acoustic properties can be promising for the establishment of standardization tool for calibration of PA, fluorescent, Raman, and other detection methods of in vivo flow cytometry and liquid biopsy.

SUBMITTER: Kozlova A

PROVIDER: S-EPMC7806591 | biostudies-literature | 2021 Jan

REPOSITORIES: biostudies-literature

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Dynamic blood flow phantom for in vivo liquid biopsy standardization.

Kozlova Anastasiia A Bratashov Daniil D Grishin Oleg O Abdurashitov Arkadii A Prikhozhdenko Ekaterina E Verkhovskii Roman R Shushunova Natalia N Shashkov Evgeny E Zharov Vladimir P VP Inozemtseva Olga O

Scientific reports 20210113 1

In vivo liquid biopsy, especially using the photoacoustic (PA) method, demonstrated high clinical potential for early diagnosis of deadly diseases such as cancer, infections, and cardiovascular disorders through the detection of rare circulating tumor cells (CTCs), bacteria, and clots in the blood background. However, little progress has been made in terms of standardization of these techniques, which is crucial to validate their high sensitivity, accuracy, and reproducibility. In the present st ...[more]

PMID: 33441866

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Project description:Pulse wave imaging (PWI) is an ultrasound imaging modality that estimates the wall stiffness of an imaged arterial segment by tracking the pulse wave propagation. The aim of the present study is to integrate PWI with vector flow imaging, enabling simultaneous and co-localized mapping of vessel wall mechanical properties and 2-D flow patterns. Two vector flow imaging techniques were implemented using the PWI acquisition sequence: 1) multiangle vector Doppler and 2) a cross-correlation-based vector flow imaging (CC VFI) method. The two vector flow imaging techniques were evaluated in vitro using a vessel phantom with an embedded plaque, along with spatially registered fluid structure interaction (FSI) simulations with the same geometry and inlet flow as the phantom setup. The flow magnitude and vector direction obtained through simulations and phantom experiments were compared in a prestenotic and stenotic segment of the phantom and at five different time frames. In most comparisons, CC VFI provided significantly lower bias or precision than the vector Doppler method ( ) indicating better performance. In addition, the proposed technique was applied to the carotid arteries of nonatherosclerotic subjects of different ages to investigate the relationship between PWI-derived compliance of the arterial wall and flow velocity in vivo. Spearman's rank-order test revealed positive correlation between compliance and peak flow velocity magnitude ( rs = 0.90 and ), while significantly lower compliance ( ) and lower peak flow velocity magnitude ( ) were determined in older (54-73 y.o.) compared with young (24-32 y.o.) subjects. Finally, initial feasibility was shown in an atherosclerotic common carotid artery in vivo. The proposed imaging modality successfully provided information on blood flow patterns and arterial wall stiffness and is expected to provide additional insight in studying carotid artery biomechanics, as well as aid in carotid artery disease diagnosis and monitoring.

Project description:BackgroundCytokine flow cytometry (CFC) or intracellular cytokine staining (ICS) can quantitate antigen-specific T cell responses in settings such as experimental vaccination. Standardization of ICS among laboratories performing vaccine studies would provide a common platform by which to compare the immunogenicity of different vaccine candidates across multiple international organizations conducting clinical trials. As such, a study was carried out among several laboratories involved in HIV clinical trials, to define the inter-lab precision of ICS using various sample types, and using a common protocol for each experiment (see additional files online).ResultsThree sample types (activated, fixed, and frozen whole blood; fresh whole blood; and cryopreserved PBMC) were shipped to various sites, where ICS assays using cytomegalovirus (CMV) pp65 peptide mix or control antigens were performed in parallel in 96-well plates. For one experiment, antigens and antibody cocktails were lyophilised into 96-well plates to simplify and standardize the assay setup. Results ((CD4+)cytokine+ cells and (CD8+)cytokine+ cells) were determined by each site. Raw data were also sent to a central site for batch analysis with a dynamic gating template. Mean inter-laboratory coefficient of variation (C.V.) ranged from 17-44% depending upon the sample type and analysis method. Cryopreserved peripheral blood mononuclear cells (PBMC) yielded lower inter-lab C.V.'s than whole blood. Centralized analysis (using a dynamic gating template) reduced the inter-lab C.V. by 5-20%, depending upon the experiment. The inter-lab C.V. was lowest (18-24%) for samples with a mean of > 0.5% IFNgamma + T cells, and highest (57-82%) for samples with a mean of < 0.1% IFNgamma + cells.ConclusionICS assays can be performed by multiple laboratories using a common protocol with good inter-laboratory precision, which improves as the frequency of responding cells increases. Cryopreserved PBMC may yield slightly more consistent results than shipped whole blood. Analysis, particularly gating, is a significant source of variability, and can be reduced by centralized analysis and/or use of a standardized dynamic gating template. Use of pre-aliquoted lyophilized reagents for stimulation and staining can provide further standardization to these assays.

Dataset Information

Dynamic blood flow phantom for in vivo liquid biopsy standardization.

Publications

Dynamic blood flow phantom for in vivo liquid biopsy standardization.

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