American Society for Cell Biology 2017
Visit our booth at ASCB to learn about the NEW Next-Generation IncuCyte® S3 Live-Cell Analysis System and the latest trends and applications in live-cell analysis.
Change can happen in an instant. Find out how the IncuCyte® S3 System, reagents and consumables make it easier than ever to visualize cell behavior and quantify cell function in real time. Derive deeper and more physiologically relevant information about your cells, plus real-time kinetic data, without ever removing your cells from the incubator.
Also, stop by for a chance to win prizes! Spin the wheel of fortune to win t-shirts, mugs, and more.
At the show, visit our poster presentation to learn about fluorescent cell-labelling strategies for live-cell analysis.
Fluorescent Cell-labelling Strategies Live-cell Analysis
Presenter: Kimberly Wicklund, Ph.D., Director of Product Management
Session date and time: Tuesday, December 5, 2017 from 12:00PM - 1:30PM
Location: Learning Center in Exhibit Halls C-E
Poster board number: B19
Authors: D.J. Trezise, H. Campwala, D. Appledorn, J. Rauch, M. Roddy & T.J. Dale
In the pursuit of more relevant and translational cell-based assays, researchers are increasingly turning to real-time live-cell analysis for their studies. Live-cell analysis provides cell biologists with greater insight and productivity by enabling quantification of cell function over hours, days and weeks via time-lapse image analysis, all automatically and within the controlled environment of the incubator. Many cell functions, such as proliferation, migration and neurite outgrowth, can be quantified in simple mono-cultures without fluorescent cell labels by analyzing features of phase-contrast images. This ‘label-free’ approach is desirable since it avoids complications associated with cell labels. However, there are limitations to what can be achieved without cell labels. For example, it is difficult to extract true cell count metrics from phase-contrast images in dense cultures, and in co-culture models cell labels are absolutely required to identify different cell types to observe their interactions. Accordingly, a range of different cell-labelling strategies for live-cell analysis have been developed. Protein-based fluorophores have been used for several decades to label cells and are ideally suited to real-time live-cell analysis. Fluorescent proteins (FPs) provide long-term and stable-cell labelling, are generally non-toxic and not depleted by repeated fluorescent excitation. By using specific targeting sequences in the expression constructs, proteins can be directed to label different cell types or organelles. Via this approach, we have developed a range of different vectors and delivery methods including nuclear-targeted FPs, cytoplasmic FPs and neuronal-specific FPs. These are packaged for delivery with either lentiviral or baculoviral (BacMam) expression constructs. Alternatively, in certain scenarios, it may be difficult or undesirable to transduce cells with fluorescent proteins. In these cases, cells can be labelled with cytoplasmic or nuclear specific fluorescent dyes. When used correctly, these chemical probes are non-perturbing to cells and produce uniform and homogeneous labelling. These dyes have very different properties and use protocols to each other, and to the protein labels, and are best aligned with different applications. In this poster, we will describe the use of these fluorescent protein and dye-based labelling strategies and how they can be used most effectively in live-cell analysis applications including cell proliferation, cytotoxicity, apoptosis, immune cell killing, and neurite outgrowth in both primary rodent and human iPSC-derived neurons.