Applications: Widefield Fluorescent Imaging

Widefield Fluorescent Imaging for Cell Morphology Study

Understanding how cells change shape in response to their environment is central to unlocking the complexities of gene expression, tissue organization, and disease progression. In a recent study titled “Regulation of chromatin modifications through coordination of nucleus size and epithelial cell morphology heterogeneity,” Alexandra Bermudez and the Neil Lin lab revealed a remarkable link between cell crowding, nuclear size, and epigenetic changes.

To capture this dynamic interplay, the researchers relied on widefield fluorescent imaging to monitor cellular and nuclear changes over time. Using a 20×/0.40 NA objective with an Etaluma microscope, the team performed high-resolution time-lapse imaging every five minutes for six consecutive days—an impressive feat that underscores the system’s ability to support long-term live-cell studies.

Widefield fluorescence provided the spatial clarity and sensitivity necessary to visualize chromatin states and track morphological heterogeneity across densely packed epithelial cells. By combining this imaging technique with automated media changes and consistent illumination, the researchers could maintain optimal culture conditions while collecting thousands of high-quality images for analysis.

Etaluma’s fluorescence microscopes are uniquely suited for these applications—compact enough to operate inside an incubator, yet powerful enough to reveal fine cellular details in real time. The integration of widefield fluorescent imaging into this study played a crucial role in documenting how mechanical and spatial constraints within a tissue environment influence nuclear architecture and, ultimately, gene regulation.

A Image of MDCK cells stained with Histone H3 Lysine 27 trimethylation (H3K27me3) illustrating its variable expression levels in different cells. Scale bar = 50 μm. B Correlation between normalized H3K27me3 expression level and nucleus area in MDCK cells. N = 461. 95% confidence interval is [−0.447, −0.194]. C Image of an E12.5 mouse epithelium stained with H3K27me3. Scale bar = 25 μm. D Correlation between normalized H3K27me3 expression and nucleus area in the mouse epithelium. N = 130. 95% confidence interval is [−0.459, −0.209]. E Image of MDCK cells stained with Histone H3 Lysine 9 acetylation (H3K9ac). Scale bar = 50 μm. F Correlation between normalized H3K9ac expression and nucleus area in MDCK cells. N = 1130. 95% confidence interval is [0.183, 0.293]. G Image of an E12.5 mouse epithelium stained with H3K9ac. Scale bar = 25 μm. H Correlation between normalized H3K9ac expression and nucleus area in the mouse epithelium. N = 713. 95% confidence interval is [0.0988, 0.241].