CV1000 Applications

Application : CV1000

• Early stage embryo
  • Early stage mouse embryo
  • Early stage chicken embryo
  • Early stage human embryo
• Brain slice
  • The cerebral cortex of chicken
  • Brain slice of mouse
• Fucci
  • Fucci
• Cancer cell
  • Cancer cell
• Primordial germ cells
  • Wide-area imaging of primordial germ cells
• Zebrafish
  • Zebrafish
• Plant
  • Arabidopsis thaliana

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Early stage embryo

Early stage mouse embryo

Long-term, 4D timelapse imaging

Following the injection of mouse embryos with mRNA, nearly 25,000 multicolor and multilayer confocal images of the embryos were acquired over 60 hour period as they developed to the blastocysts stage.Thereafter, they were transferred to a recipient mouse that gave birth to healthy pups, each of which developed normally and had full reproductive capability.This is firm evidence that long-term, multi-dimensional confocal imaging with CV1000 causes no harm to a delicate specimen such as an early stage embryo.

Data:  Kazuo Yamagata, PhD, Wakayama Lab. (Laboratory for Genomic Reprogramming), Center for Developmental Biology, RIKEN

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Early stage chicken embryo

Time lapse imaging of chicken embryo

Data:  Yukiko Nakaya, Ph.D., Laboratory for Early Embryogenesis , Center for Developmental Biology, RIKEN

Left:  Whole embryo
Center:  Image of lateral epiblast cells from the apical side
Right:  Magnified view of image (The boxed region shown in center image

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Early stage human embryo

Mikiko Tokoro, Ph.D., Noritaka Fukunaga, Ph.D. and Yoshimasa Asada, M.D., Ph.D. at Asada Institute for Reproductive Medicine won an ART award in video section at the ASRM (American Society for Reproductive Medicine) held in Boston on Oct 12-17th, 2013.

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Brain slice

The cerebral cortex of chicken

Interkinetic nuclear movement in the cerebral cortex of chicken

Interkinetic nuclear movement in the Cerebral cortex of chicken was observed for about 20 hours. Thanks to the real confocality and high-precision auto stage of the CV1000, it is possible to observe biological reactions in even thick specimen like tissue sections at a wide area with high resolution.

Data:  Yuji Watanabe, PhD, Department of Molecular Neurobiology, Graduate School of Life Sciences, Tohoku University

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Brain slice of mouse

Time-lapse imaging of neural progenitor cells during cortical development

Tangential time-lapse monitoring of all cell-cell borders, about 5 µm from the apical surface, in a cortical wall prepared from E13 Lyn-Venus transgenic mouse
Data:   Mayumi Okamoto, Ph.D., Department of Anatomy and Cell Biology, Nagoya University Graduate School of Medicine

Green:  Membrain（Lyn-GFP
Red:  Nucleus（H2B-RFP）
White arrow:  Neural progenitor cell, Yellow arrow, Blue arrow： Daughter cell
play

Imaging condition

Interval	5min
Z-sections/stack	30sections（1.2um/apart）
Imaging positions	20fields
Excitation wave	488nm, 561nm
Objective lens	40xDry

Reference:
Okamoto, M., Namba, T., Shinoda, T., Kondo, T., Watanabe, T., Inoue, Y., Takeuchi, K., Enomoto, Y., Ota, K., Oda, K., Wada, Y., Sagou, K., Saito, K., Sakakibara, A., Kawaguchi, A., Nakajima, K., Adachi, T., Fujimori, T., Ueda, M. Hayashi, S., Kaibuchi, K., Miyata, T.
TAG-1–assisted progenitor elongation streamlines nuclear migration to optimize subapical crowding. Nat. Neurosci., 16: 1556-1566 (2013) DOI: 10.1038/nn.3525

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Fucci

Time lapse imaging of Fucci

Healthy cell division of HeLa cells expressing Fucci was recorded for nearly 7 days.
By using incubator attachment, cells grew and divided as normally as being cultured in a CO₂ incubator.

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Primordial germ cells

Wide-area imaging of primordial germ cells

The process to form colonies of EG cells(a kind of iPS cell)from primordial germ cells expressing GFP of 12.5 days embryo of TG mouse was imaged for a long-time at the whole area of a culture dish(625fields).
As a result of 5 days imaging, colonies of EG cells were formed as frequently as was formed when the cells were cultured in a CO₂ incubator. With he CV1000, you can record whole area quite at ease when you don't know where to find the target, and can discover what happened from the acquired data.

Data:  Yasuhisa Matsui, PhD, Cell Resource Center, Institute of Development, Aging and Cancer, Tohoku University.

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Cancer cell

Imaging of 29F cells transfected with eGFP by using NeoFection

Floating 293F cells were transfected with eGFP by using NeoFection, a transfection accelerating agent made by ASTEC.The cells were shake-cultured over night. As a result of time lapse imaging, active movement of cells expressing eGFP inside the floating cell clusters, and structural changes in the cell wall such as ruffing were clearly observed.

Data:  ASTEC CO,LTD.

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Zebrafish

Zebrafish

White blood cells that patrol in zebrafish

White blood cell movement in zebra fish tail was continuously recorded. By rapidly capturing Z-slice images, 3D Movement of live specimen can be tracked in multi-color and with high resolution.

Data:  Dr.Philipp Niethammer（Harvard medical school Mitchison Lab)

 

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Plant

Arabidopsis thaliana

High-speed multi-field imaging

Changes in the shape of a vacuolar membrane during the germination process were continuously recorded. High-speed and multi-point time lapse imaging with the CV1000 allows accurate and high-resolution tracking of rapid changes in living organisms, something that has proven quite difficult with conventional imaging systems. By selecting the appropriate filter and pinhole size, thick samples and auto fluorescent plant cells can be clearly observed with the CV1000.

Data:  Chieko Saito, PhD, Senior Research Scientist, Molecular Membrane Biology Laboratory, Advanced Science Institute, RIKEN
(Present post : Network of Centers of Carbon Dioxide Resource Studies in Plants (NC-CARP), Laboratory of Cellular Biochemistry, Department of Biological Sciences, Graduate School of Science, The University of Tokyo)

 

Images with the same condition except for the pinhole size. Left:  25m, Right:  50um