Label-Free Longitudinal Imaging of Single Cell Drug Response with a 3D-Printed Cell Culture Platform

Area of Science:

• Biomedical imaging
• Chemical imaging
• Drug discovery

Background:

• Image-based phenotypic screening is crucial for preclinical drug discovery.
• Live-cell imaging captures dynamic cellular responses, offering advantages over static assays.
• Fluorescence microscopy has limitations in live-cell multiplexing and potential for phototoxicity.

Purpose of the Study:

• To develop a low-cost, 3D printed platform for live-cell imaging compatible with Stimulated Raman Scattering (SRS) microscopy.
• To enable real-time drug delivery and continuous monitoring of cellular changes.
• To showcase time-lapse SRS microscopy for mapping heterogeneous drug responses.

Main Methods:

• Development of a 3D printed cell culture platform for SRS microscopy.
• Implementation of real-time drug delivery and 24-hour time-lapse imaging.
• Establishment of a processing pipeline for extracting chemical and morphological features from SRS images.
• Utilizing multi-well perfusion parallelized with label-free chemical imaging.

Main Results:

• Demonstration of a low-cost, 3D printed platform enabling label-free, live-cell SRS microscopy.
• Successful real-time monitoring of biochemical and morphological changes with minimal photodamage.
• Mapping of heterogeneous, drug-induced single-cell responses over time.
• Identification of divergent cellular trajectories within populations.

Conclusions:

• Time-lapse SRS microscopy with the developed platform is a powerful tool for analyzing single-cell heterogeneity and drug responses.
• This approach accelerates phenotypic screening and offers a pathway toward personalized medicine.
• Label-free chemical imaging provides quantitative insights into dynamic cellular processes.