3i - Intelligent Imaging Innovations
Benjamin Atkinson
Workshop Room 6
AxL Cleared Tissue LightSheet - Axially Swept Light Sheet Microscopy System for Ultrafast Imaging of Cleared Specimens
AxL Cleared Tissue LightSheet (AxL CTLS) is a fully automated macro zoom microscope with high NA apochromatic objectives and dual-sided light sheet illumination for imaging whole organs to small animals. Custom-designed excitation objectives and patented axially swept light sheet microscopy (ASLM) produce an exceptionally thin, long and uniform lightsheet for large-scale high-resolution imaging. ASLM scans the light sheet in its propagation direction using high-speed remote focusing synchronized to the rolling shutter readout of an sCMOS camera at the size of the beam waist. This approach creates an exceptionally thin light sheet across a large field of view resulting in images with improved optical sectioning and signal-to-noise. This scanned sheet features a constant laser intensity across the field of view for an evenly illuminated image. The 0.14NA excitation objective creates a 2μm thin waist for superior axial resolution. AxL CTLS is designed to operate optimally across a range of 1.33 to 1.56 refractive indices ensuring compatibility across all available clearing methods. The large field of view of AxL CTLS enables ultrafast imaging of whole organs to small animals. An entire mouse can be prescanned in less than 60 seconds, imaged in 20 minutes and high resolution ASLM scanned in 9 hours revealing neuronal connectivity.
Vector3 - Total Internal Reflection Fluorescence (TIRF) and Photomanipulation Module
Vector3 is a motorized, spinning TIRF illuminator with three key imaging modalities: TIRF, photomanipulation and widefield epifluorescence. Intelligent beam steering and optical design allow for all three imaging modes to be combined in one compact device. Vector3 offers an expansive TIRF field of view (FN20) designed for modern sCMOS cameras. Spinning the excitation beam around the back aperture of the imaging objective creates an even TIRF field without shadows or polarization artifacts. A motorized scan lens corrects for sample height variation and ensures ideal TIRF illumination and photomanipulation spot size across the visible spectrum. An easy-to-use user interface in SlideBook allows users to optimize the excitation angle between TIRF and HILO for imaging thicker samples. Photobleaching, photoconversion, and photoablation events of user-drawn ROIs are easily scripted into image acquisition. In this workshop, we will highlight the large field of view and ease-of-use of Vector3 for TIRF and photomanipulation experiments.
