SPIM-OPT, high lateral resolution with the best z resolution in 3D fluorescence imaging

Light Sheet Fluorescence Microscopy (LSFM) or Selective Plane Imaging Microscopy (SPIM) are 3D imaging techniques that are becoming increasingly popular because they achieve excellent resolution at high penetration depths, while being minimally invasive. The lateral resolution of SPIM is comparable to that of a standard (epi) fluorescence microscope, while axial resolution is lower than the lateral (about a factor of 4). Optical Projection Tomography (OPT) can assume two primary modes - transmission and emission. In transmission mode, light traverses an optically cleared sample, providing general structural information. In emission mode, fluorescent labels in the sample are excited to produce specific functional information. OPT provides higher axial resolution than LSFM or SPIM, but lower lateral resolution. Our patented SPOT technology (Statistical Projection Optical Tomography) is a combination of SPIM and OPT, taking advantage of both technologies best features to produce high resolution in every dimension.

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360o illumination of the sample for stripe elimination

In Light Sheet Microscopy, illumination is perpendicular to detection. If a part of the sample is not transparent, single side illumination will create shadows and dark stripes on the image. Some setups are using multiple angle illumination, but alignment of two or more sheets is difficult and usually results in loss of axial resolution. Other setups perform measurements from two or four angles and recombine the images. Our patented SPOT technology uses single illumination (high axial resolution) combined with 500 angle detection, so the sample is illuminated and imaged from 360º.






True Isotropic Resolution

Thanks to the combination of SPIM and OPT techniques in one called SPOT we can achieve isotropic resolution in 3D fluorescence images, for all kinds of biologicalsamples.





Automatization

The QLS-scope includes automatic change of up to four air objectives, automatic change of up to eight wavelengths Lasers and the automatic change of the corresponding excitation and emission filters.





Highest Speed

We offer the fastest imaging speed possible (up to 10 volumes per second) that permits the capture of dynamic procedures inside the 3D structure.








Sample structural information add-on

QLS-scope includes the possibility to perform an OPT measurement in transmission mode, where white light is passed through an optically cleared sample, and registered at 180o from the system camera. In this way one can obtain structural information about the sample of interest and can correlate this image with the Light Sheet Fluorescence Microscopy image that contains specific information about the sample.








Moderate demand for data storage space

In Light Sheet Fluorescence Microscopy it is rather common to use a higher NA objective in order to achieve better lateral resolution. This results in a smaller FOV so, for large cleared samples, it is necessary to stitch several fields of view to reconstruct the whole sample.  The final stitching file is often rather large, resulting in an increasing demand for data storage capacity. Our patented SPOT (SPIM-OPT) technology summarizes the stitched fields of view (imaged with a high NA objective) in just 500 images from different angles, independently of the size of the picture. All this procedure is done automatically and the amount of images to be stored are always around 500. This decreases a lot the demand for data storage.

What to look for in a Light Sheet Microscope