Live Cell Morphology Assay

The label-free HoloMonitor® Cell Morphology Assay enables non-invasive live cell imaging and analysis of a wide range of morphological properties, including individual cell volume, area and thickness.

Individual Cell Morphology

Morphological changes caused by treatment or cellular transformation occur early, making kinetic cell morphology studies a significantly more sensitive tool than indirect biochemical assays to observe cellular response.

In the past, in vitro cell morphology has been difficult and time-consuming to quantify as it involves inconsistent staining and handling of complicated cell imaging software. Contrary to conventional live cell imagers, HoloMonitor acquires a time-lapse sequence of label-free quantitative phase images.

Cell-friendly Imaging

Since no phototoxic or fading fluorescent labels are required, an unlimited number of label-free and cell-friendly time-lapse images can be acquired just minutes apart. This allows HoloMonitor to non-invasively quantify treatment response of adherent cell populations, by automatically quantifying alterations in individual cell morphology.

Cell Morphology Analysis — Scatter Plots & Histograms

The HoloMonitor Cell Morphology Assay measures a wide range of morphological properties, including cell volume, area and thickness. Scatter plots help visualize differences between control and treated cells or to compare cellular characteristics at different time-points of the experiment.

Gating regions enable classification in subpopulations and identification of (rare) cells of interest, for further studies using HoloMonitor Cell Tracking. Histograms show cell population distribution of selected morphological parameter at selected time-points.

HoloMonitor cell morphology assay: cell morphology analysis using gated scatter plot

Screenshot of the HoloMonitor Cell Morphology Assay showing cells classified in subpopulations using gating regions.

Label-free Live Cell Assays

HoloMonitor offers several label-free live cell assays, ranging from cell culture quality control to single-cell tracking. After time-lapse image sequences have been recorded at selected sample positions, each of the assays may be used to provide additional results, without requiring further experiments or lab work.

For example, cell motility may be quantified together with cell morphology by tracking cells individually or by applying the HoloMonitor Cell Motility Assay, on the acquired time-lapse image sequence. As the saved image sequence is reused, no further samples are necessary. Additionally, cell proliferation data may also be generated by applying the HoloMonitor Cell Proliferation Assay, again using the previously acquired and saved image sequence.

A panel of cellular parameters, including cell morphology, proliferation, and motility, were quantitatively measured and analyzed using [HoloMonitor]. This novel and quantitative approach provides the opportunity to continuously and non-invasively study relevant cellular processes, behaviors, and characteristics of cancer cells and other cell types in a simple and integrated manner.

Jing Huang et al.

A Time-lapse, Label-free, Quantitative Phase Imaging Study of Dormant and Active Human Cancer Cells, Journal of Visualized Experiments (2018)

Some cell morphology assay parameters

Some of the morphological properties that can be analyzed using HoloMonitor.

Links to further applications based on cell morphology analysis

Mitosis Duration

  • Total mitosis time for each individual cell.
  • Graphs and data showing when each cell was in mitosis.

Cell Cycle Analysis

  • Assess cell cycle duration time.
  • Detect drug effects on cell cycle phase distribution.

Cell Death Kinetics

  • Detect early signs of drug responses before the actual cell death occurs.
  • Identify dying cells.

HoloMonitor® Kinetic Morphology Assessment

A vast line of morphological parameters can be determined during a time-lapse experiment using HoloMonitor. Like all HoloMonitor applications, morphology assessment is cell-friendly and analysed without the use of any labels or stains. Morphological parameters are, for example, useful

  • For evaluation of cell culture health and quality
  • As early and sensitive markers to detect drug effects
  • To provide information on drug mode of action
  • For identification of dying cells

Cell morphology analysis is often complemented by cell proliferation and cell motility studies to provide a complete picture.

Key References Cell Morphology

  • Quantitative evaluation of morphological changes in activated platelets in vitro using digital holographic microscopy
    Y Kitamura et al.
    Micron (2018)

    The feasibility of DHM (Digital Holographic Microscopy) in the quantitative evaluation of morphological changes in activated platelets was evaluated. Data shown implies that DHM using HoloMonitor could be a promissing method for quantitative examination of morphological changes in platelets in vitro. The publication also includes a comparision of the performance of different techniques for this purpose.

    Read the article
  • A Time-lapse, Label-free, Quantitative Phase Imaging Study of Dormant and Active Human Cancer Cells
    J Huang, P Guo and Marsha A. Moses
    JoVe (video journal) (2018)

    The video and publication demonstrate how HoloMonitor is used to conduct time-lapse and labeling-free characterizations of angiogenic and non-angiogenic human osteosarcoma KHOS cells. A panel of cellular parameters, including cell morphology, proliferation, and motility, were quantitatively measured and analyzed using HoloMonitor.

    Read the article
  • High accuracy label-free classification of single-cell kinetic states from holographic cytometry of human melanoma cells
    M. Hejna et al.
    Scientific Reports (2017)

    The authors used singel-cell tracking and machine learning to develop a robust method for label-free classification of adherent cells.

    Read the article

Label Your Tubes, Not Your Cells

— Introducing label-free live cell imaging

In this PHI webinar, our application specialist Lisa Bodily introduces the exciting field of label-free live cell imaging. We discuss live cell imaging techniques and highlight reasons to work label-free. You will learn how digital holographic microscopy visualizes your cells and get to know our HoloMonitor system. Also, our label-free applications for cell biology research are presented and some examples highlighted.

View the webinar on demand here

live cell imaging, cell culture and analysis