Usually, visualisation of morphological changes that occur during the cell cycle, apoptosis and cell death rely on the use of cell stains. However these protocols require the manipulation and interruption of normal cell culture conditions, and can themselves induce cell death.
The Phase Focus Virtual Lens® resolves these problems because it can generate very high contrast fluorescence-like images of live cells without the need for staining.
Moreover, Virtual Lens images are free from artefacts associated with conventional phase contrast techniques, and have an exceptionally even and dark background. Both of these aspects enhance image clarity, and provide improved fidelity data for subsequent thresholding, image analysis and feature extraction, and for integration with automated cell assay protocols.
In time-lapse movies, intracellular structures are routinely visualised, and all phases of mitosis may be observed during cell division.
Timelapse video of A549 cells imaged using the Phase Focus Virtual Lens® at 37°C for 20 hours, showing cell intensity increasing upon cell division, courtesy of Peter O’Toole, Joanne Marrison and Leonela Jimenez, Imaging and Cytometry, Department of Biology, University of York, Hesligton, York, Y010 5DD, UK.
The Phase Focus Virtual Lens® is currently available for cell imaging and other life science applications in an accessory that is integrated with a conventional Olympus BX41 microscope. This system therefore enables Virtual Lens imaging, as well as the full panoply of conventional methods.