Holotomography (HT) is a true paradigm shift in microscopy. This technology goes beyond the resolving capabilities of Confocal and Lightsheet imaging!
Tomocube uses a laser interferometric technique of image rendering which is broadly analogous to the use of X-rays in CT scans. Result: Time-lapse 3D Holographic Images!
Tomocube’s HT technology enable users to quantitatively and noninvasively investigate live biological cells and thin tissues in 3D over time. Holotomography reconstructs the 3D refractive index (RI) distribution of live cells and provides structural and chemical information about the cell, including dry mass, morphology, and dynamics of the cellular membrane. This can be done very fast and easy, because RI is an intrinsic optical parameter of a material and thus HT does not require any preparation on samples.
In order to image unstained biological samples, conventional methods used Phase Contrast or Differential Interference Contrast (DIC) microscopy. Resulting images were seen in two dimensions when biological samples are in reality, three dimensional (3D). Tomocube’s Holotomography (HT) solves this problem.
Just as a CT scan uses X-ray absorptivity as the imaging contrast to see inside a patient’s organs without invasive procedures, HT uses the refractive index (RI), an intrinsic optical parameter describing the speed of light passing a specific material, to visualize living cells and tissues. As light traverses through a sample, the various constituents scatter light differently based on their refractive index. By rotating an imaging beam 360° around the sample, we can capture a sequence of holograms from different angles. The resulting multiple 2D hologram images of the sample obtained from various illumination angles can be reconstructed into a 3D RI tomogram.
The sample is located on a stage between an objective lens and a condenser lens. A light source is split to follow two paths, the sample beam and the reference beam (the principle behind a Mach-Zehnder interferometer). The sample and the reference arms when combined generate a 2D hologram, which is recorded by a digital image sensor (CMOS). This reflects a QPI measurement principle of the phase shift relative to the reference beam.
The imaging beam illuminates the sample with an incident angle of 63°. The beam is rotated through 360° with respect to the optical axis. From the 48 overlapping captured holograms RI tomogram of the sample is then reconstructed.
Tomocube creates images of unprecedented clarity. Never before seen quantifiable pictures with 4 color fluorescence overlay. Ultra long time-lapse capable. Requires no sample preparation and does not harm cells!
AI-based cellular and sub-cellular segmentation, feature extraction and inference of molecular (fluorescent signal-like) information from HT images achieves high accuracy in label-free phenotypic analysis and opens up new avenues to AI-driven data interpretation. Following the successful launch of HT in the research market, Tomocube embarks on developments in diagnostics and personalized medicine applications.
Human Glial cell moving in 4D!
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