- Last Updated on Thursday, 06 February 2014 14:26
Live cell imaging
Label-free time-lapse imaging and analysis of cell cultures
How it is done
- Place the cell culture on the HoloMonitor stage.
- Select the Live Capture tab.
- If necessary, adjust the coarse focus. Fine focus is automatic.
- Enter the time-lapse duration and the time interval between each image frame.
- Start image acquisition.
- View the time-lapse in the View Images tab.
- Optionally select the Export Images tab to create a time-lapse video.
The HoloMonitor time-lapse cytometers facilitate long-term, label-free, live cell imaging and quantification of cellular dynamics.
Optionally, recorded time-lapse image sequences are automatically processed to give cytometric data such as cell count, cell morphology and cell motility at any point in time.
Stains and fluorescent labels are toxic to cells and may even cause DNA damage
(Ge et al
J. Ge, D. K. Wood, D. M. Weingeist, S. Prasongtanakij, P. Navasumrit, M. Ruchirawat and B. P. Engelward
'Standard fluorescent imaging of live cells is highly genotoxic',
Cytometry Part A, vol. 83A, no. 6, pp. 552–560, 2013 ). To reduce the risk of inducing artifacts, cells should be studied without using stains or labels whenever possible.
During the few milliseconds it takes the HoloMonitor to record an image, the observed cells are illuminated with a dimmed, unfocused, red laser beam. As cells are imaged unstained, the translucent cells only absorb extremely small amounts of light energy. This makes phototoxicity effects very unlikely, as no energy is transferred to the cells that can cause phototoxicity.
The non-invasive nature of HoloMonitor time-lapse cytometers make them ideal for label-free live cell imaging assays, stretching over days. Adherent cells are imaged and quantified directly in the cell culture vessel, leaving cells completely undisturbed.
Contrary to conventional microscopes and image cytometers, the HoloMonitor records a hologram. From the hologram a viewable image is created by the software, at an automatically selected focal distance. This enables the HoloMonitor to automatically compensate for mechanical focus drift. When a drift occurs, a slightly different focal distance is selected when creating the viewable image. Holographic images can in fact be refocused at any time after the have been recorded.
For long-term live cell imaging experiments, the HoloMonitor M3 can be combined with a standard microscopy stage incubator or heated plate from Okolab and others. The HoloMonitor M3 is compatible with microfluidics solutions adapted for live cell imaging, provided by IBIDI, CellAsic and others. The compact HoloMonitor M4 is small enough to be placed in any standard cell incubator.