Discriminating different grades of cervical intraepithelial neoplasia based on label-free phasor fluorescence lifetime imaging microscopy

Abstract

This study proposed label-free fluorescence lifetime imaging and phasor analysis methods to discriminate different grades of cervical intraepithelial neoplasia (CIN). The human cervical tissue lesions associated with cellular metabolic abnormalities were detected by the status changes of important coenzymes in cells and tissues, reduced nicotinamide adenine dinucleotide (phosphate) (NAD(P)H) and flavin adenine dinucleotide (FAD). Fluorescence lifetime imaging microscopy (FLIM) was used to study human cervical tissues, human cervical epithelial cells, and standard samples. Phasor analysis was applied to reveal the interrelation between the metabolic changes and cancer development, which can distinguish among different stages of cervical lesions from low risk to high risk. This approach also possessed high sensitivity, especially for healthy sites of CIN3 tissues, and indicated the dominance of the glycolytic pathway over oxidative phosphorylation in high-grade cervical lesions. This highly adaptive, sensitive, and rapid diagnostic tool exhibits a great potential for cervical precancer diagnosis.

FLIM as a Promising Tool for Cancer Diagnosis and Treatment Monitoring

“…Reproduced with permission from Ref. [ 56 ]. Copyright 2020, Optical Society of America; a-iii NADH/NAD + ratio is applied for ascertaining tumor fronts.…”

“…Furthermore, for clinical diagnosis, Phasor-FLIM has been successfully applied to cervical intraepithelial neoplasia (CIN) and neoplastic skin lesions, as shown in the phasor plot in Fig. 3a-ii [ 56 ]. This method is also adopted to glioblastoma multiforms (GBMs), leukemia subpopulation classification (Fig.…”

“… Reproduced with permission from Ref. [ 56 ]. Copyright 2020, Optical Society of American; b Scatter plot of 65 WBCs (blue), 35 Jurkat cells (orange), 46 K562 cells (green), and 35 THP-1 cells (cyan) based on NADH fluorescence signals.…”

Leveraging Methylation Alterations to Discover Potential Causal Genes Associated With the Survival Risk of Cervical Cancer in TCGA Through a Two-Stage Inference Approach

“…Family with sequence similarity 73, member A (FAM73A) is the downregulated gene of DNA from exfoliated cervical cells in terms of the HPV-16 variant analysis (Green, 2019;Meng et al, 2020). CRYZL1 contains a reduced nicotinamide adenine dinucleotide (phosphate) (NAD(P)H) binding site which is involved in cellular metabolism, while cervical lesions are associated with cellular metabolic abnormalities (Wang et al, 2020). It is previously found that the members of the ZNF family interact with nucleic acids, proteins, and small molecules and are involved in a variety of crucial molecular processes in cervical tumor cells at replication, transcriptional, and translational levels.…”

Microscopic phase reconstruction of cervical exfoliated cell under partially coherent illumination

“…Cervical cancer is one of the most common malignant tumors. Early diagnosis and early treatment are important for patients' prognoses [1][2][3]. Cervical cancer can be prevented through Human Papillomavirus (HPV) vaccine injection (primary prevention) and HPV DNA detection cervical exfoliated cell cytological screening (secondary prevention).…”