A novel combined fluorescent probe staining method for circulating tumor cell identification

Area of Science:

• Biomedical diagnostics
• Cancer research
• Molecular imaging

Background:

• Circulating tumor cells (CTCs) are crucial biomarkers for cancer diagnosis and prognosis.
• Accurate identification of CTCs remains a challenge in clinical practice.
• Novel methods are needed to enhance CTC detection sensitivity and specificity.

Purpose of the Study:

• To develop and validate a novel, highly accurate method for identifying circulating tumor cells (CTCs).
• To assess the utility of this new method in cancer diagnostics and prognostics.
• To compare the performance of the new method against established techniques.

Main Methods:

• Developed and optimized a combined fluorescent probe staining protocol using three probes (Dil, Hoechst 33342, PY).
• Validated the protocol using in vitro tumor cell lines and 32 patient tumor samples.
• Compared results with Wright's Giemsa staining and CD45 immunocytochemistry (ICC).

Main Results:

• The combined fluorescent probe staining method demonstrated comparable morphology to Giemsa staining in vitro.
• In vivo analysis showed differences in cell diameter and nucleo-cytoplasmic ratio compared to other methods.
• Achieved a higher CTC detection rate (14/32) and AUC (0.844) with lower standard deviation.

Conclusions:

• A novel combined fluorescent probe staining method offers higher accuracy for CTC identification than Wright's Giemsa staining.
• This technique is proposed as a valuable tool for clinical cancer diagnostics and prognostics.
• CTCs identified by this method showed correlation with platelet levels.