Livecyte — not another microscope!

ATA Scientific Pty Ltd
Thursday, 04 June, 2020

Reveal complex individual cell behaviours and unlock unique insights in every assay.

Live-cell time-lapse microscopy is an established and powerful technique for the study of mammalian cell biology in vitro. Multiple microscopy technologies exist each presenting their own set of benefits and limitations.

Individual cell segmentation and tracking using traditional label-free methods such as brightfield or phase contrast is challenging due to lack of inherent imaging contrast. Fluorescent labels enhance cell contrast but also have the potential to alter normal cell function and induce toxicity. The high-intensity light required to excite fluorophores can also alter cell behaviour and induce cell death largely due to photodamage. A consequence of this is the subtle changes in cell morphology, motility and proliferation that may have unforeseeable effects on experimental outcomes that are often overlooked.

Why aren’t conventional systems delivering?

The loss of true data is increasingly normalised; subtle phenotypical changes are lost due to deficient modalities forcing constant compromises to gain contrast. Throughput often trumps detail! In an attempt to create a stable environment of temperature and humidity, some microscopes are subjected to a life in an incubator, leaving them prone to mould and degradation of electronic componentry leading to unanticipated repair costs. Microscopy seems to have moved from an investigatory tool to a mass screening machine. Remember — the devil is in the detail!

So how do we do this better?

Ideally, live-cell imaging needs to identify and track individual cells for prolonged periods without the need for perturbing labels and provide high-contrast images under low levels of light intensity, to preserve natural behaviours and allow recovery of cells for subsequent experimentation or downstream analysis.

The ability to segment and track individual cells and their generational lineages is paramount for accurate quantification of cell behaviour. A continuous, large field of view with no loss of resolution or focus that permits even highly motile cells to be tracked during time-lapse imaging can prevent potentially important cells from being lost or overlooked.

Information-rich reliable data is key where each experiment automatically yields a plethora of phenotypic parameters such as cell thickness, volume, dry mass in addition to kinetic behaviour characterised by cell speed, displacement and confinement ratio. Imaging systems should be easy to use, require no calibration and no dedicated consumables, and have no hidden costs.

Phasefocus Livecyte delivers all of this!

Phasefocus Livecyte generates high-contrast, fluorescent-like images, using low-powered illumination (4–7 µW/mm2) in which cells appear as bright objects on a dark background. The enhanced contrast in combination with phase retrieval data increases the robustness of single-cell segmentation and tracking algorithms without the need for dyes or probes. This form of quantitative phase imaging (QPI), ptychography, is an emerging imaging technique that retrieves phase-delay of light passing through a cell. Livecyte can provide you with data not available with any other instrument.

How can you extract more knowledge from your assays?

Livecyte can extract the changes in morphology, motion and dry mass of each cell over time. This leads to a more complete characterisation of cell phenotypic properties. Tracking and analysis of individual cells, along with population metrics, to monitor cell speed and directionality of migration together with cell proliferation can allow greater insights into biological processes. Livecyte offers the versatility to measure and monitor sensitive cell types such as primary cells, patient-derived cells and stem cells. These types of cells are much closer to their natural origins compared with immortalised cell lines, providing a more realistic account of cell behaviour in response to treatment conditions. Livecyte can also perform correlative fluorescence and brightfield imaging.

Dry mass — a class above confluence

You know from your cultures that cells spread out, ball up and grow without dividing, and their division is not always symmetrical. Given confluence simply measures the change in plate coverage by cells; relying on this rudimentary metric alone clearly results in unacceptable misleading outcomes. Dry mass is the summed mass of all cellular components excluding water. As such, the dry mass measurement is an accurate measure of cell size, accounting for the extent of biosynthetic and degradative processes in addition to uptake and expulsion material by the cell.

Achieve more from one experiment

Livecyte enables a vast array of metrics to be calculated and combined to perform a number of applications such as true proliferation, advanced scratch wound, cell motility, mitotic time and morphology. Within each dashboard application there are a wealth of outputs. Imagine this kind of depth of analysis for every dashboard, for every well, for every cell, for every experiment.

Only one thing to do!

Contact us at ATA Scientific and discover more.

For a clearer version of the top image, click here.

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