Project
409-Ichimura-AMATERAS
Title
Ultra-large FOV imaging by AMATERAS-2
Description
The authors established a volumetric trans-scale imaging system with an ultra-large field-of-view (FOV) that enables simultaneous observation of millions of cellular dynamics in centimeter-wide three-dimensional (3D) tissues and embryos. Using a custom-made giant lens system with a magnification of ×2 and a numerical aperture (NA) of 0.25, and a CMOS camera with more than 100 megapixels, they built a trans-scale scope AMATERAS-2, and realized fluorescence imaging with a transverse spatial resolution of approximately 1.1 µm across an FOV of approximately 1.5×1.0 cm2. The 3D resolving capability was realized through a combination of optical and computational sectioning techniques tailored for our low-power imaging system. They applied the imaging technique to 1.2 cm-wide section of mouse brain, and successfully observed various regions of the brain with sub-cellular resolution in a single FOV. They also performed time-lapse imaging of a 1-cm-wide vascular network during quail embryo development for over 24 hr, visualizing the movement of over 4.0×105 vascular endothelial cells and quantitatively analyzing their dynamics. Their results demonstrate the potential of this technique in accelerating production of comprehensive reference maps of all cells in organisms and tissues, which contributes to understanding developmental processes, brain functions, and pathogenesis of disease, as well as high-throughput quality check of tissues used for transplantation medicine.
Funding
Grant-in-Aid for Scientific Research on Innovative Areas “Singularity Biology (No. 8007)” 21H00431 (YS), 18H05416 (HH), 18H05412 (SO), and 18H05410, 18H05408 (TN)
Grant-in-Aid for Transformative Research Areas (A) “Seeing through Scattering Media (No. 20A207)” 21H05590 and 23H041350 (TI)
the Research Program of "Five-star Alliance" in "NJRC Mater. & Dev." (TN)
Precursory Research for Embryonic Science and Technology (PRESTO) JPMJPR18G2 (TI) JSPS KAKENHI JP23H00395 and JP24K22022 (HH)
AMED Brain/MINDS JP21dm0207117 (HH) and
BINDS JP23ama121054 and JP23ama121052 (HH)
The Uehara Memorial Foundation (TN) Takeda Science Foundation (TN, HH)
Core Research for Evolutionary Science and Technology (CREST) JPMJCR15N3 (TN), JPMJCR1926 (KS, SO)
RIKEN Cluster for Science, Technology and Innovation Hub (SO)
JST NBDC Grant Number JPMJND2201 (SO)
Title
Z-series images of mouse brain stained with SYTOX-Green
Description
Z-series SYTOX-Green images of mouse brain obtained by AMATERAS2. The colonally cutted 1.5 mm-thick brain section was chemically cleared using CUBIC, and the cell nuclei were stained with SYTOX-Green. The images were obtained with 2×magnification system, and the background intensity was removed by the computational sectioning.
Description
The fixed and cleared mouse brain block followed by staining with SYTOX-Green.
Gene ontology: Biological processes
Gene ontology: Cellular components
Gene ontology: Molecular functions
Dimensions
13264x9180x378x1x1
Detection methods
complementary metal oxide semiconductor (CMOS)
Visualization methods
SYTOX Green
Illumination methods
complementary metal oxide semiconductor (CMOS)
Contrast enhancing methods
computational method
Resolution enhancing methods
Sample preparation methods
formaldehyde fixed tissue
Light source
Thorlabs SOLIS-470C
Detector
CIS, VCC-120CXP1M
Filter set
Edmund Optics, #87–800 and #86–992
Dichroic
custom-made (cut-off: 497 nm)