MuVI SPIM

A major goal in the field of neuroscience is to understand the connectivity of the brain across scales, from individual synaptic connections to whole organism connectomics. Visualizing the whole brain often requires imaging at depths that are not supported by conventional imaging methods, such as laser scanning confocal microscopy. In recent years, light-sheet fluorescence microscopy (LSFM) has emerged as an imaging technique capable of addressing a wide variety of applications in neuroscience due to its faster acquisition speed and larger imaging depth compared to confocal microscopy. Although early implementations of LSFM were optimized around imaging developmental processes in typical model organisms, technological advances in optics have allowed researchers to extend this technique to much larger samples, such as an entire mouse brain. Combined with emerging tissue clearing techniques, researchers are now able to investigate intact tissues, organs, and even entire organisms with subcellular resolution using light microscopy methods. The versatility of LSFM for neuroscience research has contributed to its rapid adoption and increasing popularity in the scientific community. However, there are multiple important factors one must consider when moving from standard imaging techniques to LSFM. In this application note, Bruker discusses the reasons why LSFM and clearing techniques are particularly useful for neuroscience research, important considerations for neuroscientists adopting this technology, and application examples of LSFM for cleared sample imaging.

Using conventional light microscopy techniques to image large or dense biological samples can be challenging due to light scattering properties of tissues and mismatched refractive indices. The combination of tissue clearing and light-sheet microscopy enables the fast, gentle, and high-resolution imaging deep within thick samples. Here, Bruker Fluorescence Microscopy presents a recap of the webinar presented by Sharla White, Ph.D., Vice President of Research and Development at ClearLight Bio, and the practical applications and best-known methods of combined light-sheet fluorescence microscopy (LSFM) and CLARITY tissue clearing are presented. An example of CLARITY tissue clearing on mouse intestines is also presented, showcasing how imaging with LSFM provides added insight to the unique structure, shape, and layers of the intestine.

Bruker's Luxendo light-sheet technology can image a wide range of live, fixed, and cleared biological samples from organoids and embryos to large whole organisms. Overcoming many of the barriers that conventional light microscopy faces, lightsheet fluorescence microscopy, also called selective plane illumination microscopy (SPIM), supports advanced research in the life sciences. SPIM works by de-coupling the fluorescence excitation and detection beams and together with a sheet of light for excitation, selectively illuminates a focal plane for high-resolution images without the danger of phototoxicity. Here, Dr. Martin Jechlinger, the Senior Scientist and Head of VISION Laboratory at the MOLIT Institute, talks about how his lab utilizes light-sheet microscopy to investigate 3D organoid cultures. Specifically, the molecular mechanisms underlying mammary tumor development during breast cancer, as well as what is happening when treatments are failing.

Imaging living plants involves challenges on several levels, including sample properties (e.g., aerial and roots, chlorophyll, cell walls), tissue visualization (e.g., dyes vs reporter lines), sample-friendly imaging (e.g., embedding, low phototoxicity, timelapses), and efficient data handling (e.g., processing and storage). Working together with end users and application specialists, Bruker Luxendo has the experience needed to deliver expert commercial LSFM solutions that can significantly aid the imaging of living and naturally growing plants. In this application note, Bruker highlights challenges with using microscopy for image acquisition of plants, challenges specific to LSFM, data processing challenges, and how Bruker Luxendo can support you in overcoming these obstacles to create meaningful data.