Time-lapse Imaging Cytometry

Time-lapse imaging cytometry by holographic microscopy allows non-invasive visualization and analysis of live cell populations by tracking and quantifying individual cells.

Understanding Cellular Dynamics

An advantage of holographic microscopy is that the created quantitative phase images are focused when viewed, not when recorded. This makes the HoloMonitor time-lapse cytometer ideal for long-term imaging and analysis of living cells by means of time-lapse micro­scopy, which acquires a series of cell images at regular time intervals to analyze the dynamics of various cellular events. Un­focused images, caused by focus drift, are simply refocused by letting the computer soft­ware recreate the phase image from the recorded hologram.

A time-lapse microscopy image showing a Monster HeLa cell, acquired by the HoloMonitor imaging cytometer.

An example time-lapse image sequence of a monster HeLa cell, imaged by HoloMonitor. HeLa cells are cancer cells named after Henrietta Lacks, who in the early 1950s donated the first cells that was successfully kept alive and cultured in a laboratory environment. HeLa cells and other immortal cells are today routinely cultured by scientists to study the complex behavior of cells and their response to drug treatments.

In addition to identifying each individual cell, HoloMonitor provides data for analysis of more than 30 morphological parameters. However, the true power of time-lapse cytometry first emerges when the same cells are monitored over time. The HoloMonitor design utilizes recent tech­nological advances to allow time-lapse image sequences of cultured cells to be effortlessly recorded over long time periods.

With HoloMonitor installed in a cell incubator, the cells are kept in a cell friendly environment during the entire experiment. Long-term live cell kinetic data can easily be obtained using time-lapse microscopy. Images are recorded at selected intervals, down to 1 image/sec. Depending on the app­lication, cell images are played back as a video recording to aid analysis of dynamic cell behavior.

A time-lapse image sequence of a wound healing scratch assay, acquired by holographic time-lapse microscopy using the HoloMonitor imaging cytometer.

Time-lapse image sequence of a wound healing assay experiment.

Cytometric Imaging Software

From recorded time-lapse image sequences, the Holo­Monitor App Suite software helps the user to auto­matically extract and kinetically analyze live cell population data based on individual cell data. When preferred, individual cell data — such as cell count, cell morphology, cell velocity, and cell division rate — can all be measured from the same experimental time-lapse data, without requiring additional experiments.

HoloMonitor App Suite software

HoloMonitor App Suite

Time-lapse Cytometry References

  • 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
  • HoloMonitor M4: holographic imaging cytometer for real-time kinetic label-free live-cell analysis of adherent cells
    Mikael Sebesta; Peter J. Egelberg; Anders Langberg; Jens-Henrik Lindskov; Kersti Alm; Birgit Janicke
    Proceedings, Quantitative Phase Imaging II (2016)

    Live-cell imaging enables studying dynamic cellular processes that cannot be visualized in fixed-cell assays. An increasing number of scientists in academia and the pharmaceutical industry are choosing live-cell analysis over or in addition to traditional fixed-cell assays. We have developed a time-lapse label-free imaging cytometer HoloMonitor M4. HoloMonitor M4 assists researchers to overcome inherent disadvantages of fluorescent analysis, specifically effects of chemical labels or genetic modifications which can alter cellular behavior. Additionally, label-free analysis is simple and eliminates the costs associated with staining procedures.

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  • Moving into a New Dimension: Tracking Migrating Cells with Digital Holographic Cytometry in 3D
    A. Görloff Wingren
    Cytometry Part A (2018)

    Commentary article discussing the fundamental role of cell movement studies in cancer research. Advantages of the HoloMonitor Cell Tracking and Wound Healing Applications over the transwell migration and invasion assays are highlighted, and includes the possibility to use the cells in experiments for other purposes after completing the imaging. In addition to single cell tracking, the HoloMonitor technology also benefits from the fact that morphology analysis can be performed of each cell. The author concludes that this indeed opens up for almost unlimited possibilities to perform cell morphology analysis using this methodology, since each image is very rich in cellular information.

    Read the article
More Cell Imaging and Cytometry
An apoptotic cell explosion imaged by the HoloMonitor live-cell microscope.
Gentle Live Cell Imaging
Sophisticated live cell imaging techniques enable non-invasive analysis of cell populations on a single-cell level.
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Quantitative Phase Imaging
Quantitative phase imaging (QPI) provides both quantitative and beautiful images of living cells, transforming phase micro­scopy into a quantitative tool for detailed live cell analysis.
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A quantitative phase image of a cell and the hologram it was created from
Holographic Microscopy
Holographic microscopy is the most common form of quanti­tative phase imaging. The HoloMonitor live cell time-lapse cytometers employ holographic micro­scopy to allow non-invasive visualization and quanti­fication of living cells without compro­mising cell integrity.
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