Dataset Information

Label-free detection and size estimation of combustion-derived carbonaceous particles in a microfluidic approach.

ABSTRACT: Detection and size estimation of combustion-derived carbonaceous particles (CDCPs) are important to understand their toxicity. Size determination of individual nano- and microparticles (NMPs) based on scattered light is a straightforward method. However, detection and sizing of CDCPs in biological samples based on scattering alone are not possible due to the compositional heterogeneity of NMPs present in biological samples. Label-free identification of CDCPs based on unique white light (WL) emission, using femtosecond (fs) pulsed near-infrared (NIR) lasers, has emerged as a reliable method even in complex biological samples. However, size estimation of CDCPs in biological samples using label-free techniques is still lacking. Here we report the development of a dual-channel multiphoton flow cytometry (DCMPFC) setup for label-free identification and size-determination of CDCPs in suspensions. Scattering intensity calibration with reference polystyrene (PS) nanoparticles (NPs) and Mie Theory allow us to determine the sizes of CDCPs in aqueous suspensions. Further, the relationship between particle sizes and WL emission intensity was determined, and the sizes of CDCPs in urine samples could also be estimated. This approach is believed to open new opportunities for the quantification and size determination of CDCPs, originating from exposure to air pollution, in liquid biopsies. This is an important step in determining the CDCP exposure of individual persons.

SUBMITTER: Aslam I

PROVIDER: S-EPMC9419628 | biostudies-literature | 2022 Jul

REPOSITORIES: biostudies-literature

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Label-free detection and size estimation of combustion-derived carbonaceous particles in a microfluidic approach.

Aslam Imran I Fron Eduard E Roeffaers Maarten B J MBJ

Nanoscale advances 20220621 15

Detection and size estimation of combustion-derived carbonaceous particles (CDCPs) are important to understand their toxicity. Size determination of individual nano- and microparticles (NMPs) based on scattered light is a straightforward method. However, detection and sizing of CDCPs in biological samples based on scattering alone are not possible due to the compositional heterogeneity of NMPs present in biological samples. Label-free identification of CDCPs based on unique white light (WL) emis ...[more]

PMID: 36132818

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Project description:Cancer cells metastasize through the bloodstream either as single migratory circulating tumor cells (CTCs) or as multicellular groupings (CTC-clusters). Existing technologies for CTC enrichment are designed primarily to isolate single CTCs, and while CTC-clusters are detectable in some cases, their true prevalence and significance remain to be determined. Here, we developed a microchip technology (Cluster-Chip) specifically designed to capture CTC-clusters independent of tumor-specific markers from unprocessed blood. CTC-clusters are isolated through specialized bifurcating traps under low shear-stress conditions that preserve their integrity and even two-cell clusters are captured efficiently. Highly parallel architecture of the chip allows deterministic screening of clinically relevant volumes of blood samples at slow, and hence, non-damaging flow rates. Using the Cluster-Chip, we identify CTC-clusters in 30-40% of patients with metastatic cancers of the breast, prostate and melanoma. RNA sequencing of CTC-clusters confirms their tumor origin and identifies leukocytes within the clusters as being tissue-derived macrophages. Together, the development of a device for efficient capture of CTC-clusters will enable detailed characterization of their biological properties and role in cancer metastasis. We used the Cluster-Chip to capture CTC-clusters from the blood of a breast cancer patient with high CTC counts, released CTC-clusters in solution, stained them with TexasRed-conjugated antibodies against the leukocyte cell surface markers CD45, CD14 and CD16, and then isolated intact CTC-clusters individually using a micromanipulator. From a single time point, we retrieved 15 CTC-clusters, and each of those clusters was individually subjected to RNA-sequencing analysis using a next generation platform (SOLiD 5500). In addition, two leukocytes were isolated from the blood of a healthy donor were individually subjected to RNA-sequencing analysis using the same platform.

Dataset Information

Label-free detection and size estimation of combustion-derived carbonaceous particles in a microfluidic approach.

Publications

Label-free detection and size estimation of combustion-derived carbonaceous particles in a microfluidic approach.

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