3D Fluorescence Imaging Added to Holotomography Microscopy for World First

The HT-2 from Tomocube is the world’s first microscope to combine both holotomography and 3D fluorescence imaging into one unit. Capable of simultaneously capturing high resolution 3D optical diffraction tomography and 3D fluorescence images, the new microscope enables long-term tracking of specific targets in live cells while minimising stress. The capability to easily deliver holotomography and fluorescence correlative analysis in 2D, 3D and 4D will enable researchers and clinicians to open new frontiers in bioscience and better understand, diagnose, and treat disease.

“Research and diagnosis in the life sciences depends on the information that can be found using cellular analysis,” says Aubrey Lambert, Tomocube’s Chief Marketing Officer. “Current microscopy techniques limit the quantity and quality of information available to researchers and clinicians and harm the living cells during long-term studies, but the HT-2 is set to overcome these limitations.”

The new microscope builds on the novel holotomographic technology first shown in the Tomocube HT-1, providing morphological, chemical and mechanical properties of cells through the 3D refractive index (RI) tomograms but adding molecular specificity information through the fluorescence imaging capability. The highly detailed fluorescence images show the position of specific target organelles or structures in living cells, while consecutive measurements of time-lapse 3-D RI tomography enables the monitoring of cells and their structures with minimal stress.

The HT-2 incorporates a customisable three-channel LED light source (385, 470 and 570 nm) and a motorized Z-drive with a step resolution of 150 nm to generate highly detailed Z-stack images. The TomoStudio software suite controls all the functionality of the HT-2 and provides fast imaging capability and 2D/3D/4D visualization of the cellular images based on 3D RI distributions of the cells and tissues. At its core is the same patented complex digital micromirror device (DMD) optical light shaper used in the Tomocube HT-1. Consisting of several hundred thousand micromirrors arranged in a rectangular array, the DMD eliminates the need for moving parts in the lightpath and delivers stable performance during long-term studies.

Benefits:

Correlative microscopy in one instrument - HT-2 provides high-quality 3D images of both holotomography and 3D fluorescence for each sample.

Quantitative data marked with fluorescence - HT-2 provides morphological (volume, surface area, projection area, sphericity and ellipticity), chemical (dry mass, concentration) and mechanical (cell deformability) properties of cells with 3D refractive index (RI) tomogram. Moreover, fluorescence image provides information about molecular specificity.

Live cell molecular and holographic imaging with minimal stress on cells - Simultaneous measurement capability of time-lapse 3-D RI tomography and fluorescence image allows long-time tracking of specific targets in live cells. The fluorescence image provides the position of specific target organelles or structures in live cell, and consecutive measurements of time-lapse 3-D RI tomography enables the monitoring of cells and their structures with minimal stress.