Organoids, lab-grown mini-organs derived from stem cells, mimic the key functional, structural, and biological complexities of real organs and can be used to unravel biological mechanisms, model disease, and test treatments. One of the challenges of researching organoids is obtaining fast and sensitive images of their three-dimensional structure and the dynamics within. Standard confocal microscopy, a widely used approach for visualizing fluorescent molecules in cells and tissues, has limitations in speed and signal-to-noise ratio, as well as being phototoxic to cells over longer periods of imaging.
A new technique, line rescan confocal imaging, promises to overcome these limitations. NL5+, developed by Confocal.nl, is a fast line rescanning confocal system with high sensitivity and resolution which provides clear images even from the dimmest samples while exposing the cells to very low laser powers. NL5+ works by moving a line of laser light across the sample, rather than a single laser point as in laser scanning confocal microscopy (LSCM). This makes it possible to obtain images faster, without loss of resolution or contrast. In addition, NL5+ can use up to five lasers, allowing the user to be flexible while looking deeper into the sample.
Line rescan microscopy with NL5+ is excellent for biological applications where a combination of speed and high sensitivity is required. For example, it is possible to study organoids from different organs, such as the intestine, liver, kidney, and brain. NL5+ can provide information on morphology, cell differentiation, gene expression, signal transduction, and interactions of organoids with their environment. This can lead to new insights into the development, function and pathology of organs, and to new possibilities for personalized medicine.
Line rescan microscopy is a simple, robust, and versatile technique that is easy to use. It is possible to convert almost any existing wide-field fluorescence microscope into a fast and sensitive 3D confocal imaging system with the addition of NL5+ module. This way one obtains a future-proof system for studying organoids and their application like the optimization of organoid culture conditions to better predict clinical outcomes during new drug development.