Confluency and cell count are critical parameters when sub-culturing cells. However, assessing these values can be highly subjective and error-prone, leading to inconsistent experimental outcomes. The Millicell® Digital Cell Imager (Millicell® DCI) standardizes determination of common cell culture parameters and enables in-vessel analysis to conserve precious sample. Additionally, the Millicell® DCI benefits from cloud-based capabilities for off-instrument visualization, storage, and analysis of captured images.
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In this podcast, Kimberly Italiano, product manager for the Millicell® Digital Cell Imager, explains how digital cell imagers provide more consistent cell culture as well as other benefits.
For cultured cells to remain healthy and viable, they must be sub-cultured into fresh vessels. Determining when to perform sub-culture has traditionally been based on a visual estimation of confluency, yet this approach is prone to inter- and intra-operator variability that can have undesirable consequences. While delayed passaging can allow cell overgrowth that may lead to altered cellular characteristics (e.g., stem cells may lose their differentiative potential), premature sub-culture can extend workflows and delay time to results, which can incur unexpected costs.
After estimating confluency, researchers typically perform a cell count. Manual cell counting involves removing a sample from the culture vessel, diluting it as necessary, and pipetting it onto a hemocytometer; a manual clicker is then used to tally cells within a counting grid using a microscope and the estimated total cell count in the culture is calculated. As well as being both tedious and time-consuming, manual cell counting introduces further sources of variability, with each of the steps just described being subject to user error. Moreover, the length of time spent in suspension prior to transfer to a new culture vessel can be detrimental to certain cell types.
By providing objective determination of common cell culture parameters including confluency, cell count, and morphology, digital cell imagers automate the decision on when to perform sub-culture. Automation standardizes cell culture protocols to ensure more consistent experimental outcomes and simplifies analysis, freeing up researchers’ time to be spent on other tasks. Notably, more consistent cell culture practices help preserve key cell characteristics such as phenotype and specific metabolic rate (SMR) that cannot be controlled by standardized seeding densities alone.
A further important benefit of digital cell imagers is that they enable storage and retrieval of archival data and images. Not only does this improve traceability, but it also presents opportunities to monitor growth trends over time. The latter can especially be helpful when working with difficult cultures, to determine whether a culture is growing abnormally, or to predict when cultures will be ready for seeding, transfection, induction, harvest, or any other form of manipulation.
The Millicell® DCI is an intuitive digital imaging platform designed for automated analysis during routine culture of adherent cells—including stem cells, spheroids, and organoids. Using the Millicell® DCI, researchers can perform measurements directly in the culture vessel to both conserve sample and decrease handling time, thereby mitigating the risk of sample contamination, and can efficiently achieve reproducible analysis of confluency with minimal hands-on time required.
In addition to measuring confluency, the Millicell® DCI generates an estimated adherent cell count (through in-vessel analysis) as well as enables a total cell count to be performed using a hemocytometer. In either scenario, the illumination, focus, Z-axis position, and magnification can all be adjusted using the built-in touchpad display, giving the end user complete control without loss of objectivity. Critically, automated software performs all mathematical calculations to reduce error.
Another key feature of the Millicell® DCI is that it permits the creation of individual user profiles, each with its own customizable settings. This provides researchers with rapid access to pre-set analytical parameters based on project or cell type and allows data to be annotated for easier retrieval and sorting downstream. For those working with unusual cell types and cultures, including spheroid and organoid cultures, specialized software algorithms are available to streamline analysis.
With wireless transfer capabilities to the cloud, the Millicell® DCI enables data to be sorted, viewed, and re-analyzed via a web-based application. The optional cloud-based subscription service can be used to save costs and support data redundancy for backup and protection, helping safeguard the integrity and longevity of research projects.
To learn more about the Millicell® DCI and how it can standardize routine cell culture for more consistent experimental outcomes, visit millicelldci.com