High‐Viability Circulating Tumor Cells Sorting From Whole Blood at Single Cell Level Using Laser‐Induced Forward Transfer‐Assisted Microfiltration

The efficient isolation and molecular analysis of circulating tumor cells (CTCs) from whole blood at single-cell level are crucial for understanding tumor metastasis and developing personalized treatments. The viability of isolated cells is the key prerequisite for the downstream molecular analysis, especially for RNA sequencing. This study develops a laser-induced forward transfer -assisted microfiltration system (LIFT-AMFS) for high-viability CTC enrichment and retrieval from whole blood. The LIFT-compatible double-stepped microfilter (DSMF), central to this system, comprises two micropore layers: the lower layer's smaller micropores facilitate size-based cell separation, and the upper layer's larger micropores enable liquid encapsulating captured cells. By optimizing the design of the DSMFs, the system has a capture efficiency of 88% at the processing throughput of up to 15.0 mL min<sup>-1</sup> during the microfilter-based size screening stage, with a single-cell yield of over 95% during the retrieval stage. The retrieved single cells, with high viability, are qualified for ex vivo culture and direct RNA sequencing. The cDNA yield from isolated CTCs surpassed 4.5 ng, sufficient for library construction. All single-cell sequencing data exhibited Q30 scores above 95.92%. The LIFT-AMFS shows promise in cellular and biomedical research.

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Publication Details

Journal
Advanced Science
Published
2025-01-27
DOI
https://doi.org/10.1002/advs.202414195
Citations
4
Primary Topic
Cancer Cells and Metastasis
Type
article
Field-Weighted Citation Impact
4.18

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article

High‐Viability Circulating Tumor Cells Sorting From Whole Blood at Single Cell Level Using Laser‐Induced Forward Transfer‐Assisted Microfiltration

Zhenhe Xu, Yuntong Wang, Fu-Long Ma, Ying Xue et al.
4 citations
Advanced Science
Cancer Cells and Metastasis
4.18
article

High‐Viability Circulating Tumor Cells Sorting From Whole Blood at Single Cell Level Using Laser‐Induced Forward Transfer‐Assisted Microfiltration

Zhenhe Xu, Yuntong Wang, Fu-Long Ma, Ying Xue, Yingcun Xia, Songtao Dou, Xu Han, Kunlong Zhang, Bei Li, Yi Zhang, Wei Wang, Hang Li, Huan Wang, Yang Xu, Qi Wang, Qingmei Xu, Xiao Ma
article en
4 citations

Abstract

The efficient isolation and molecular analysis of circulating tumor cells (CTCs) from whole blood at single-cell level are crucial for understanding tumor metastasis and developing personalized treatments. The viability of isolated cells is the key prerequisite for the downstream molecular analysis, especially for RNA sequencing. This study develops a laser-induced forward transfer -assisted microfiltration system (LIFT-AMFS) for high-viability CTC enrichment and retrieval from whole blood. The LIFT-compatible double-stepped microfilter (DSMF), central to this system, comprises two micropore layers: the lower layer's smaller micropores facilitate size-based cell separation, and the upper layer's larger micropores enable liquid encapsulating captured cells. By optimizing the design of the DSMFs, the system has a capture efficiency of 88% at the processing throughput of up to 15.0 mL min<sup>-1</sup> during the microfilter-based size screening stage, with a single-cell yield of over 95% during the retrieval stage. The retrieved single cells, with high viability, are qualified for ex vivo culture and direct RNA sequencing. The cDNA yield from isolated CTCs surpassed 4.5 ng, sufficient for library construction. All single-cell sequencing data exhibited Q30 scores above 95.92%. The LIFT-AMFS shows promise in cellular and biomedical research.

Advanced ScienceVol. 12(18)
East China University of Science and Technology (CN), Dalian Medical University (CN), Chinese Academy of Sciences (CN), Peking University (CN), Changchun Institute of Optics, Fine Mechanics and Physics (CN), Second Affiliated Hospital of Dalian Medical University (CN), Taiyuan Institute of Technology (CN), University of Chinese Academy of Sciences (CN), Beijing Advanced Sciences and Innovation Center (CN)
National Natural Science Foundation of China, Peking University, National Key Laboratory of Science and Technology on Micro/Nano Fabrication
Openalex Percentile: Top 7%
Cancer Cells and Metastasis
4.18
DOIOpen Access PDFOpenAlex
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