Light sheet fluorescence microscopy (LSFM) has emerged as a powerful tool for fast and gentle imaging of whole living model organisms, tissues, and cells as they develop over extended periods. The ZEISS Lightsheet 7 takes this technology a step further, enabling the imaging of large optically cleared specimens in toto with exceptional subcellular resolution. This cutting-edge system is designed with dedicated optics, sample chambers, and holders that can be adapted to the refractive index of various optical clearing methods, ensuring compatibility with a wide range of tissue types, fluorescent labels, and sample sizes.

With high quantum efficiency detectors, Lightsheet 7 allows researchers to observe even the fastest biological processes with minimal illumination, thereby reducing light-induced phototoxicity and preserving the biological integrity of live samples. This functionality is bolstered by a specialized sample chamber that can maintain optimal conditions through heating, cooling, and CO2 regulation, creating the perfect environment for a variety of experiments.

Lightsheet 7 supports imaging of specimens up to 2 cm in size at refractive indices between 1.33 and 1.58, accommodating almost all clearing solutions. This versatility enables scientists to acquire detailed overview images and data with subcellular resolution, whether they are working with optically cleared organoids, spheroids, organs, or complex neuronal structures. Additionally, the system’s smart software tools facilitate the adjustment of imaging parameters, including light sheet and sample positions, zoom settings, and data processing, while the patented Pivot Scan technology ensures the capture of artifact-free optical sections with outstanding image quality. In essence, ZEISS Lightsheet 7 represents a significant advancement in LSFM, expanding its application potential across a diverse range of biological research scenarios.

Sample Preparations:

Lightsheet 7 is a typical orthogonally arranged SPIM. In such a system, the sample is Vertically upright and remains stationary within a glass capillary tube, embedded in a rigid Medium such as agarose.

Sample preparations require the following components:

a) Sample: Samples can be either live or fixed, depending on the requirements of the experiment. They should be transparent, either naturally or through a pre-treatment, to allow the light sheet to penetrate effectively. The size of the sample should fall within the millimeter range, specifically measuring 1/3 to 2/3 of the diameter of the glass capillary.

b) Rigid Medium: In order to effectively utilize Light Sheet Fluorescence Microscopy (LSFM) for imaging whole living model organisms, tissues, and cells, it is crucial to maintain a stable environment for the sample. For this purpose, a rigid yet porous medium is required to hold the sample in place while also facilitating nutrient exchange for live samples. Agarose or agars, with their suitable properties, have proven to be ideal mediums for this requirement. A commonly used solution is 0.5-1% filtered low-melting agarose, which effectively immobilizes the sample while enabling the necessary exchange.