Exploring tumour heterogeneity in lung cancer and responses to cancer drugs using semi-quantitative phase contrast microscopy

Poster presented as part of the Crick BioImage Analysis Symposium 2023.

Heterogeneity is a key feature of many tumours, including Non-Small Cell Lung Cancer (NSCLC), and plays a vital role in therapy resistance and tumour relapse. Little is known about how individual tumour cells within the heterogenous mixture may be sensitive or resistant to targeted treatments and how they may evolve over time. Here we use single cell resolved time-lapse imaging tools to follow the fate of individual tumour cells during targeted therapy, coupled with the use of fluorescent biosensors as a readout of MAPK pathway activity. We describe a novel label-free microscopy technique for single-shot, semi-quantitative phase microscopy, polarisation differential phase contrast (pDPC),implemented into an existing fluorescence microscope. We use pDPC microscopy for whole cell segmentation, generation of several cell morphological features as well as cell dry mass. We track these metrics over time, in addition to the readout of MAPK pathway dynamics at the single cell level, to probe responses targeted inhibitors. Our results demonstrate that using pDPC is comparable to using fluorescent labels for single cell segmentation and subsequent cell tracking and can be combined with fluorescence to study how targeted inhibitors affect the inter-connectivity between MAPK dynamics, cell migration, and cell mass in NSCLC.

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