Dr. Tom Villani from Visikol and Dr. Ann Rossi and Hilary Sherman from Corning published an article in Labcompare that discusses the challenges associated with imaging 3D cell culture models and extracting quantitative data.
With the widespread adoption of 3D cell cultures as more physiologically relevant in vitro models, there has arisen a need to adjust the methods of characterizing these models that may differ from standard 2D cell cultures. Neuronal spheroids are particularly challenging to image due to their dense nature and thus labeling methods need to be specifically optimized for their characterization.
Thermo Fisher Scientific recently launched the CX7 LZR HCS confocal system which has been integral to Visikol’s success in offering advanced 3D cell culture assays to its clients. The CX7 LZR system is ideal for 3D cell culture analysis as it is powered by seven solid state lasers which provide the ability to image 3D tissues past a depth of 1 mm when combined with Visikol® HISTO-M™ tissue clearing.
In order to maintain specimen integrity, one can utilize the high signal to noise benefits of fluorescent labeling along with tissue clearing and confocal microscopy to image full volumes of tissue to characterize blood vessels in three dimensions. Indeed, although the vasculature of various tissue types can form complex networks of interconnectivity, the labeling can be accomplished rather simply through a variety of techniques, ranging from simple use of highlighter ink[ii] to more specific immunolabeling such as with CD31[iii]. The technique implemented depends on the desired measurables, which may be as basic as total visualization of tissue vasculature, or quantitative comparison of functioning versus non-functioning blood vessels.
To help with slicing mouse brains into 1 mm thick sections, we have launched 1 mm coronal and sagittal mouse brain slicers which are inexpensive, easy-to-clean and easy to use. These slicers allow for rapid and easy tissue clearing and 3D tissue imaging.
In their pursuit of cutting edge technologies and creative new ideas, Visikol has built a STEM internship program with Somerville High School where students work on the Visikol team on real-world projects.
Thermo Fisher Scientific recently announced the launch of their CytoVista™ tissue clearing reagents which are designed for use with both 3D cell culture models as well as whole tissues. The CytoVista™ tissue clearing workflow is compatible with most fluorophores, including fluorescent proteins that are detected with common fluorescent imaging instruments, such as wide-field microscopes, high content analyzers, confocal microscopes, and lightsheet microscopes