Cell Cycle Analysis - Mitotic Cells in HeLa Cultures
Classification of cell cycle stages in HeLa cells using morphology-based analysis with only standard nuclear staining. This dataset demonstrates identification of cell cycle phases (G0/G1, S/G2, M) using nuclear morphology without specialized dyes. Successfully isolated 164 mitotic cells (3%) from ~5,500 total cells through sequential gating based on nuclear characteristics.
Methodology
HeLa cells cultured in 8-well ibidi slides and fixed using 4% PFA for 15 min. Cells stained with Hoechst 33342 (nuclear stain) and Phalloidin-Alexa Fluor 488 (actin cytoskeleton). Images acquired using Nikon TI2 widefield microscope with DIC.
Usage examples
Complex cell cycle analysis without specialized dyes (e.g., EdU, BrdU). Retrospective analysis of existing datasets. Sequential gating to isolate mitotic cells with high purity. Morphological profiling of cell cycle stages.
Technical notes
Key morphological features used: Area, Circularity, Mean DAPI Intensity, Major Axis Length, Minor Axis Length. Final gating based on nuclear elongation (Major vs. Minor Axis Length) was critical for removing contaminating interphase cells. Distribution: G0/G1 (65%), S/G2 (23%), Mitosis (3%), Other/Debris (9%).
Assays in this dataset
Cell Health & Morphology
Cell Morphology & Phenotypic Profiling
Measures the size, shape, and fluorescence intensity of individual cells across multiple markers.
Cell Health & Morphology
Cell Counting & Viability
Counts total cells and classifies them as live or dead based on marker intensity, providing the most fundamental readout for toxicity and dose-response experiments.
Cell Health & Morphology
Cell Cycle Analysis
Categorizes cells into G0/G1, S/G2, and M phases using cell morphology and DNA stain intensity, preserving spatial context unlike flow cytometry.
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