Optical thin section microscopy has rapidly gained popularity in volume imaging due to three key characteristics: firstly, by limiting excitation to near the focal plane, light damage is minimized, for example, biological survival time is longer. Secondly, obtaining good optical sections, usually close to confocal microscopy. Thirdly, the collection speed is very fast, several orders of magnitude faster than traditional confocal microscopes. The main drawback of SPIM is that it requires additional optical components to generate the light sheet. The most common method is to place a separate illumination objective orthogonal to the detection objective and place the optical device that produces the paper between the illumination objective and the laser source. Adding additional lenses will impose space limitations on the imaging system and sample installation. Essentially, microscopes need to be designed around the sample, so there are various types of optical microscope designs, each of which is most suitable for different samples and installation requirements. In contrast, traditional confocal or epifluorescence microscopes only have one optical path and can accommodate a wider variety of samples. In other words, the advantage of SPIM is that it comes at the cost of narrower applicability for any individual tool.

Place two objective lenses at a right angle above the sample horizontally mounted in an open culture dish, with each objective lens at a 45 degree angle to the vertical direction. Create a light sheet from one objective lens and image it using another objective lens. Collect a pile of images by moving the light sheet through the sample. For some applications, 3D information in a single view or stack is sufficient (iSPIM). For a dual view system, the two objectives act in opposite directions to collect another stack from a vertical direction, and then the two datasets can be computationally merged to generate a 3D dataset with isotropic resolution (overcoming the problem of typical axial resolution differences to obtain information from other views). Therefore, the dual view diSPIM has two (usually symmetrical) optical paths, including two scanners and two cameras.
The diSPIM "head" can be installed on various inverted microscopes, including the RAMM frame of ASI. The diSPIM system can be obtained from various system integrators. Various open-source and proprietary software packages are available for data collection and processing. Regardless of the system integrator and software used, most underlying microscope hardware is the same.
The selection of diSPIM targets is limited as they must focus together without colliding with each other. The most commonly used objective for diSPIM is a 40x water immersion objective with an NA of 0.8 (Nikon CFI Apo 40XW NIR). Olympus 20x/0.5 objective lens is another possibility. 1) Nikon 10x/0.3. ASI and Special Optics have jointly developed a transparent tissue objective suitable for diSPIM, which can image transparent tissue up to 5 mm deep in flat form or in a 12 mm spherical capsule. Single sided systems (iSPIM) have greater flexibility as the illumination objective can be a low NA long WD objective. SCMOS cameras are most commonly used for SPIM imaging. The diSPIM system is equipped with Hamamatsu Flash4, Andor Zyla, PCO Edge, and Photometrics Prime 95B cameras. ASI has manufactured compact fiber coupled 2D galvanometers or 'scanners', which are components of the system. The original version of the scanner creates a light sheet by quickly scanning on one axis and moving the light sheet through sample 2) using another axis. We also offer a scanner version with cylindrical lenses for generating static light sheets. The output of the excitation laser (or laser emission) is simply fed into the scanner. Using a 2x1 optical switch or dual output laser emission is very helpful, so that the excitation can be fully guided to the scanner in the active optical path.
For applications that require environmental control, diSPIM can easily be equipped with a thermostatic enclosure and appropriate equipment to maintain the survival and happiness of the samples. Bottom objective lenses (inverted microscopes) typically have lower magnification objectives and cheaper cameras for locating samples. It is easy to add drop lighting.

Like other optical imaging techniques, diSPIM only illuminates the focal plane, making it an ideal choice for imaging live cells and organisms, as it minimizes photobleaching and phototoxic effects to the greatest extent possible. Compared with traditional or rotating disk confocal systems, the axial resolution has been increased by about 2 times, photobleaching has been reduced by more than 10 times, and the speed is comparable to that of rotating disks. View a more detailed comparison with Confocal. Compared to many other optical implementations, the main advantage of diSPIM is that, similar to inverted microscopes, sample installation is very simple. The most common method is to place the specimen on a 24 x 50 mm cover glass, which is fixed in a special chamber that can accommodate the immersion medium. Other lamps with open installation do not have isotropic resolution. View a more detailed comparison of the light film method. In addition to facilitating sample positioning, the bottom objective lens can also be used for light manipulation (including optogenetics) or other experimental techniques. It can also be used to provide a third independent view of the sample. This flexibility is also a unique advantage of diSPIM. DiSPIM is a modular microscope that allows for multiple variations and additions based on your specific needs. NIH researchers, Applied Scientific Instrumentation, and others are exploring various new features and improvements. Please refer to the partial variant list. The diSPIM system can be purchased from multiple system integrators. Compared to other commercial lightweight sheet solutions, they are cheap and flexible. Compared to customized SPIM/LSFM systems, they are easy to obtain, use, and maintain. Please also refer to the more comprehensive technical comparison of SPIM technology.

ASI provides all necessary hardware to implement diSPIM, a flexible and easy-to-use selective planar illumination microscope (SPIM) implementation that enables dual view (d) of the sample when installed upside down (i). Microscope. The diSPIM "head" can be installed on various inverted microscopes, including the RAMM frame of ASI. ASI manufactures optomechanical components, including electric platforms, 2D galvanometers for creating and moving optical sheets, and piezoelectric objective lens movers. We need objective lenses, lasers, and cameras to complete the system; Users can purchase other items themselves, use the services of various system integrators who sell diSPIM, or purchase them through ASI. DiSPIM has successfully tested the cells cultured on the cover slip and embedded in the cell c on the college gel. Embryos of nematodes and zebrafish, as well as many other samples.