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<p>Circulating tumor cells (CTCs) are important tumor metastasis and recurrence biomarkers. However, their low prevalence and variability pose significant challenges to achieving clear morphological observations and accurate molecular characterization. Hence, CTCs were isolated from the peripheral blood of 25 non-small cell lung cancer (NSCLC) patients using microfluidic chip. To enhance tumor cell identification, giemsa staining was employed to visualize the morphological features of isolated CTCs, followed by multiple immunofluorescence staining targeting specific tumor markers for confirmatory analysis. Integrating these two staining methods improved the accuracy of CTC identification and solved problems such as nonspecific binding that are common when using a single staining technique. Additionally, a significant correlation between CTC count and cancer progression was observed, with higher CTC content in advanced cancer stages. The cascade staining methodology elevates CTC identification efficacy, thereby facilitating dynamic monitoring of lung cancer progression through sequential morphological and molecular characterization.</p>

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Development and Application of Cascade Staining for the Morphological and Phenotypic Characterization of Circulating Tumor Cells

  • 期刊论文

  • 发表日期：

    2026年4月10日

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  • Chunyan Wang,
  • Xiaodong Ren,
  • Huimin Chong,
  • Jinmi Li,
  • Xiange Sun,
  • Hong Wang,
  • Leilei Wei,
  • Qing Huang

摘要

Circulating tumor cells (CTCs) are important tumor metastasis and recurrence biomarkers. However, their low prevalence and variability pose significant challenges to achieving clear morphological observations and accurate molecular characterization. Hence, CTCs were isolated from the peripheral blood of 25 non-small cell lung cancer (NSCLC) patients using microfluidic chip. To enhance tumor cell identification, giemsa staining was employed to visualize the morphological features of isolated CTCs, followed by multiple immunofluorescence staining targeting specific tumor markers for confirmatory analysis. Integrating these two staining methods improved the accuracy of CTC identification and solved problems such as nonspecific binding that are common when using a single staining technique. Additionally, a significant correlation between CTC count and cancer progression was observed, with higher CTC content in advanced cancer stages. The cascade staining methodology elevates CTC identification efficacy, thereby facilitating dynamic monitoring of lung cancer progression through sequential morphological and molecular characterization.