Dataset Fig4_Hoechst

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Project

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.
License
CC BY
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)

Dataset

Title
Z-series images of myocardial organoids stained with Hoechst33342
Description
Z-series images of myocardial organoids by AMATERAS2. The organoid was stained with Hoechst33342 and immunostained with troponin T. This dataset includes the images of Hoechst33342 The data of immunostaining with troponin T is reposited in another dataset, Fig4_Troponin.
License
CC BY
Submitted at
None
Released at
None
Updated at

Biosamples

Description
Myocardial organoids derived from hiPSC stained with anti-cardiac troponin T with Alexa488, and Hoechst33342.
Organisms
Homo sapiens (NCBI:txid9606)
Strains
Cells
pluripotent stem cell (CL_0002248)
Cell lines
hiPSC line 201B7 ()
Intrinsic variables
Extrinsic variables

Ontologies

UBERON
Anatomical entities
Gene ontology: Biological processes
Gene ontology: Cellular components
nucleus (GO:0005634)
Gene ontology: Molecular functions

Genetics

Gene names
Protein names
Genetic methods
Protein tags
Probes
Hoechst33342
Oligo Primer
Genotype

Treatments

Reagent or Compound
Concentrations
FoldDilution

Imaging Methods

Dimensions
19568x12588x75x1x1
X scales
0.75 micrometer
Y scales
0.75 micrometer
Z scales
4 micrometer
T scales
Channels
1 channel
Microscopy types
fluorescence microscopy (FBbi:00000246) (AMATERAS-2w)
Detection methods
complementary metal oxide semiconductor (CMOS)
Visualization methods
Hoechst 33342
Illumination methods
complementary metal oxide semiconductor (CMOS)
Sources of contrast
Contrast enhancing methods
computational method
Resolution enhancing methods
Image parameters
Sample preparation methods
formaldehyde fixed tissue

Instruments

Body
custom-made
Model
Light source
Thorlabs SOLIS-385C
Detector
CIS, VCC-250CXP1M
Objective
custom-made
Filter set
Edmund Optics, #87–800 and #86–992
Dichroic
custom-made (cut-off: 497 nm)