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Scientists from the University of Leuven (KU Leuven), and VIB, a life sciences research institute based in Belgium, have developed a multi-color optical method for 3D imaging surfaces and structures.
The research paper published in the BMC explains, “Current mesoscale 3D imaging techniques are limited to transparent or cleared samples or require the use of X-rays. This is a severe limitation for many research areas, as the 3D color surface morphology of opaque samples cannot be assessed.”
ALMOST - Label-free - Multicolor - Optical - Surface
Dubbed as “ALMOST” (A Label-free Multicolor Optical Surface Tomography), this method can be used for digital repositories of zoological and botanical collections which can then be 3D printed.
Micro and mesoscale 3D imaging have allowed for the analysis of microorganisms, embryos, and organs within biomedical research.
Sebastian - Munck - Microscopy - Expert - VIB-KU
Nevertheless, according to Sebastian Munck, a light microscopy expert from VIB-KU Leuven, 3D imaging requires making a sample transparent using chemical ‘clearing’ methods. Such methods are “time intensive and can’t be applied to every type of sample.”
“MOREOVER, IF YOU WANT TO STUDY SURFACE MORPHOLOGY OR COLOR, OPTICALLY CLEARING IS COUNTERPRODUCTIVE.”
Order - Image - Details - Samples - Researchers
Thus, in order to 3D image morphological details of non-transparent samples, the researchers combined optical projection tomography (OPT) and oblique illumination – a technique whereby light is projected at an angle to reveal features with higher contrast. Furthermore, color filters and a projection algorithm were used together with 3D rendering software to eliminate the need for clearing methods.
“ALMOST opens the possibility for longitudinal imaging of unaltered, live samples,” the study states.
Information - Transmission - Fluorescence - Approach - OPT
“It reveals complementary information to transmission and fluorescence, it poses an ideal supplementary approach to well-established OPT and light sheet modalities and allows imaging of the sample color, which is lost in X-ray-based techniques like micro-CT.”
The oblique illumination/imaging chamber for reflected light imaging. It is crucial to use a reflective chamber, lined with white paper, to promote diffuse illumination. Image via VIB-KU...
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