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CSU-X1, CSU22, CSU10 Applications

Application : CSU-X1, CSU22, CSU10


Vesicle trafficking

Vesicle Trafficking Vesicle Trafficking Vesicle Trafficking Vesicle Trafficking

*Movies from Indiana Center for Biological Microscopy, taken by Kenneth Dunn, Ph.D. (movies of Vesicle trafficking)

All images were collected using the system described below:
Nikon TE 2000U inverted microscope, using Nikon 60 or 100 NA 1.4, oil-immersion plan apochromatic objectives, with a Hamamatsu Orca-ER CCD system

GFP labeled tubulin in living Madin-Darby canine kidney cells

GFP labeled tubulin in living Madin-Darby canine kidney cells
play  (832 KB)

GFP labeled Rab7 and GFP labeled tubulin microtubules in living Madin-Darby canine kidney cells

GFP labeled Rab7 and GFP labeled tubulin microtubules in living Madin-Darby canine kidney cells, 380 frames collected at 18fps, movie plays back at 2x speed
play  (1,409 KB)

GFP labeled tubulin in living Madin-Darby canine kidney cells

GFP labeled tubulin in living Madin-Darby canine kidney cells, images collected at 2fps for 60 seconds, movie plays back at 15x speed
play   (909 KB)

GFP labeled Rab7 and GFP labeled tubulin in living Madin-Darby canine kidney cells

GFP labeled Rab7 and GFP labeled tubulin in living Madin-Darby canine kidney cells
play   (1,867 KB)

GFP labeled tubulin in living Madin-Darby canine kidney cells

GFP labeled tubulin in living Madin-Darby canine kidney cells, rendered using our Voxx software
play  (3,988 KB)

GFP labeled tubulin in living Chinese Hamster Ovary Cells,

GFP labeled tubulin in living Chinese Hamster Ovary Cells, rendered using our Voxx software
play (4,809 KB)


Cell Division

Cell Division Cell Division Cell Division Cell Division
Cell Division Cell Division Cell Division Cell Division

Cell Division of a LLCPK1 cell

*Movies from Patricia Wadsworth’s laboratory

Cell Division of a LLCPK1 cell expressing GFP-tagged EB1, a microtubule plus-end binding protein (kind gift of Lynne Cassimeris).


(All movies were taken by Dr. U. Serdar Tulu by using MicroMax camera on a Nikon TE-300 inverted microscope coupled to a Yokogawa spinning disc confocal unit (Perkin Elmer Life & Analytical Sciences. )and are shown at the speed of 10 frames/sec.)

cell1

Interphase-1 movie:
The cell is in G1/S phase, which is characterized by presence of one centrosome. Images were recorded every 4 seconds for a total of 3 min with 600 msec exposure-time
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cell2

Interphase-2 movie
The cell is in G2 phase, which is characterized by presence of two centrosomes. Images were recorded every 4 seconds for a total of 3 min with 600 msec exposure-time
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cell3

Metaphase movie:
Images were recorded every 2 seconds for a total of 2 min with 600 msec exposure-time.
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Cell Division

*Movies by Drs. Jonathan M. Scholey & David J. Sharp

Interphase S2 cell expressing EGFP-tubulin

Interphase S2 cell expressing EGFP-tubulin
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Mitotic S2 cell expressing EGFP-tubulin

Mitotic S2 cell expressing EGFP-tubulin 
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Mitotic S2 cell expressing EGFP-EB1

Mitotic S2 cell expressing EGFP-EB1 
play    (765 KB)

Drosophila syncytial blastoderm-stage embryo injected with rhodamine tubulin and expressing GFP-histone

Drosophila syncytial blastoderm-stage embryo injected with rhodamine tubulin and expressing GFP-histone. 
play    (10,210 KB)

Intravital Molecular Imaging

Movies by Paul Maddox, Ludwig Inst. of Cancer Research Time-lapse images were acquired by using a CSU10 Yokogawa spinning-disk confocal system (Perkin-Elmer Life Sciences Wallac) mounted on a Nikon Eclipse TE300 inverted microscope (Nikon Instrument Group). The specimens were illuminated at 488 nm with an air-cooled Ar/Kr laser (60mW, Melles Griot). Digital images were acquired with an Orca ER,Hamamatsu 16-bit cooled CCD camera (Hamamatsu Photonics), providing the goa-1 and gpa-16 DNA constructs, Stephan Grill and the acquisition system was controlled by MetaMorph software (Universal Imaging). Fluorescence images were acquired with 350 ms exposure at 2- or 5-s intervals by using a Plan Apochromat 100_/1.4 NA objective and 2 _ 2 binning in the camera. Images were analyzed by using MetaMorph software and Microsoft Excel (Microsoft) and were processed with Adobe Photoshop (Adobe Systems)

Drosophila embryo expressing GFP-centromere protein & injected with X-rhodamine tubulin

Drosophila embryo expressing GFP-centromere protein & injected with X-rhodamine tubulin , Maddox et.al. Current Biology(2002)
play     (770 KB)

GFP-Histone & GFP spindle poles in C. elegans embryoGFP-Histone & GFP spindle poles in C. elegans embryo

GFP-Histone & GFP spindle poles in C. elegans embryo.
play     (803 KB)

GFP-Histone & GFP spindle poles in C. elegans embryo

GFP tubulin in C. elegans embryo.
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GFP-Histone & GFP spindle poles in C. elegans embryoGFP-Histone & GFP spindle poles in C. elegans embryo

Hela_yfp_cenpa_control1 : HeLa cell expressing YFP-CENP-A(kinetochore specific protein) with DIC. 
play  (575 KB)

C. elegans embryo

*Movies by Fumio Motegi & Asako Sugimoto
(Laboratory for Developmental Genomics, RIKEN Center for Developmental Biology)

cell1

Dynamics of GFP-EBP-1 (a homolog of EB1, a microtubule plus-end binding protein) during the first cell division of a C. elegans embryo. Images were acquired with a 1-s exposure time at 2-s intervals.
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cell2

Dynamics of GFP-RHO-1 at the cell cortex during the establishment of anterior-posterior polarity in a C. elegans one-cell embryo. Images were acquired with a 0.5-s exposure time at 5-s intervals. The embryo is oriented with the anterior pole to the left.
play  (14,464 KB)

Tobacco BY-2 cell

*Movies by Takumi Higaki & Seiichiro Hasezawa
Laboratory of Plant Cell Biology in Totipotency,
Department of Integrated Bioscience, Graduate school of Frontier science,The University of Tokyo

Cell division of tobacco BY-2 cell

Cell division of tobacco BY-2 cell
Red:  chromosome(RFP-HistoneH2B)
Green:  microtubule(GFP-tubulin),
Blue:  cell plate(FM4-64)
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Cell division of tobacco BY-2 cell

Localization of actin microfilaments
Left:  Control
Right:  BA treatment
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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

Blue:  Neural progenitor cell(M phase), Magenta:  Daughter cell

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


FRET Imaging

Real Time/Real Color FRET Imaging Real Time/Real Color FRET Imaging Real Time/Real Color FRET Imaging Real Time/Real Color FRET Imaging

Real-time/real-color FRET imaging/Yellow Cameleon

Takeharu Nagai, and Atsushi Miyawaki A real-time movie of confocal real color images and corresponding pseudocolored ratio images showing Ca2+ waves inside cells, which were evoked by histamine, in HeLa cells expressing YC3.60(Yellow Cameleon). The images were taken at video rate (30 Hz) using a color 3CCD camera for simultaneous acquisition of CFP (cyan-emitting mutant of GFP) and YFP (yellow-emitting mutant of GFP) images. To improve z axis resolution, a spinning disk confocal unit was placed in front of the camera. Ten micromolar histamine was added to the recording medium to activate receptor-evoked Ca2+ release from endoplasmic reticulum.

Left:  A real-time movie of confocal real color images
Right:  Corresponding pseudocolored ratio images
play  (672 KB)

Real-time/real-color FRET imaging/Yellow Cameleon

A real-time movie of confocal images and corresponding pseudocolored ratio images showing Ca2+ waves inside cells, which were evoked by histamine, in HeLa cells expressing YC3.60(Yellow Cameleon). 

FRETFRET

Left:  A real-time movie of confocal image of YFP channel
Right:  Corresponding pseudocolored ratio images
play  (19,925 KB)


Calcium Imaging

Real Time Calcium Imaging Real Time Calcium Imaging Real Time Calcium Imaging Real Time Calcium Imaging

Real-time intracellular calcium dynamics of the Purkinje fibers in the perfused rat heart

*Movies from Tetsuro Takamatsu’s laboratory (In Japanese)
The fluo3-fluorescence (8 bits, pseudo-color image) of the perfused rat heart visualized by the CSU-based real-time confocal microscopy at 30 frames/s. 
Pixels 205 x 230. The scale bar denotes 50 m.
Microscope: BX50WI, Olympus、Objective lens: LUMIPlan FL x20, Olympus, with CSU-21, 
CCD camera: ES310 Turbo, Roper Scientific & Image intensifier: VS4-1845, Videoscope.

Spatially uniform Ca2+ transients are identified in Purkinje fibers on the ventricular septal wall.

Spatially uniform Ca2+ transients are identified in Purkinje fibers on the ventricular septal wall.
play   (3,731 KB)

Simultaneous imaging of phosphatidyl inositol metabolism and Ca2+ levels in PC12h cells

BBRC 308 (2003) 673-678
Mitsuhiro Morita, Fumito Yoshiki, and Yoshihisa Kudo
Laboratory of Cellular Neurobiology, School of Life Science, Tokyo University of Pharmacy and Life Science (in Japanese)
Relationship between phosphatidyl inositol metabolism and Ca 2+ level was analyzed with high spatial resolution at video rate in PC12h cells double-labeled with GFP fused with the pleckstrin homology domain and FuraRed,
CSU10, C6790 CCD camera and W-View; simultaneous dual color imaging system (Hamamatsu), Nikon E600FN microscope with Fluor 40X/0.8W.

Simultaneous imaging of phosphatidyl inositol metabolism and Ca2+ levels in PC12h cells

Left:  EGFP
Right:  FraRed
play ( 3,731 KB)

Functional multineuron calcium imaging

Yuji Ikegaya
Laboratory of Chemical Pharmacology, Graduate school of Pharmaceutical Sciences, The University of Tokyo.
fMCI is a functional imaging technique with multicell loading of calcium fluorophores. fMCI has unique advantages, including: i) recording en masse from hundreds of neurons in a wide area, ii) single-cell resolution, iii) identifiable location of neurons, and iv) detection of non-active neurons during the observation period

Simultaneous imaging of phosphatidyl inositol metabolism and Ca2+ levels in PC12h cells

Spontaneous firing-induced somatic calcium spikes of CA3 pyramidal cells in a rat hippocampal slice culture loaded with Oregon Green 488 BAPTA-1AM

High-speed calcium imaging

Data:  Yuji Ikegaya Ph.D., Graduate School of Pharmaceutical Sciences at the University of Tokyo

Cell assemblies in spontaneous activity of a CA3 neuron population

Cell assemblies in spontaneous activity of a CA3 neuron population
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Clustered synaptic inputs in a spontaneously active CA3 neuron

Clustered synaptic inputs in a spontaneously active CA3 neuron
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in vivo imaging

in vivo imaging in vivo imaging

Intravital Molecular Imaging

Satoshi Nishimura
Department of Cardiovascular Medicine, Graduate School of Medicine, The University of Tokyo
Dr. Satoshi Nishimura is a world-renowned expert of intravital molecular imaging; which means in vivo imaging of microcirculation and molecular dynamics in living animals with high spatiotemporal resolution.

Intravital Molecular imaging is a powerful tool to elucidate not only vascular pathological conditions such as arteriosclerosis, but also to study molecular mechanisms of pathological conditions caused by complicated and multi-cellular abnormal interactions, such as cancer or metabolic syndrome. 

He has developed imaging system based on highly spatiotemporal imaging with the CSU and visualized microcirculation, vascular functions and multi-cellular dynamics of target molecules in real-time and in threedimensions, in vivo.

Using this technique, he successfully elucidated how obese adipose tissue played a bad role in obesity. 

Intravital Molecular Imaging

An example of real-time movie of microcirculation in mouse, which clearly shows dynamic movement and interactions among leucocytes, platelets, macrophages and endothelium.
play (10,706 KB)

Intravital Molecular Imaging

Cellular kinetics in microcirculation
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Intravital Molecular Imaging

Deformed erythrocyte and platelets in microcirculation
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Long-time, 4 colors, and 3D in vivo imaging.

Mikala Egeblad, Andrew Ewald, Zena Werb., UCSF, Dept of Anatomy

Researchers at UCSF developed in vivo imaging system capable of multicolor, 3D imaging at multi-points to observe tumor microenvironment in the same live mouse for long time, by using the CSU.

Long-time, 4 colors, and 3D in vivo imaging

Nuclear EGFP in Foxp3+ Tregs in a tumor within a live MMTVPyMT; ACTB-ECFP; Foxp3EGFP mouse.
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Long-time, 4 colors, and 3D in vivo imaging

Membrane EGFP in CD11c+ dendritic-like cells in a tumor within a live MMTVPyMT; ACTB-ECFP; CD11-DTR-EGFP mouse.
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Long-time, 4 colors, and 3D in vivo imaging

Example of migration of two myeloid cells at the border of a late carcinoma – a low-migratory, dextran-ingesting cell and a high-migratory, dextran-negative cell.
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Long-time, 4 colors, and 3D in vivo imaging

Dynamics of several types of stromal cells in different tumor microenvironment for up to 12 hours to find different behaviors of each stromal cell
Green:  c-fms-EGFP
Blue:  ACTB-ECFP
Red:  10 kDa dextran
play


Multi-color imaging

Multicolor livecell imaging Multicolor livecell imaging

Multi-color imaging of live cells and tissues

Satoshi Nishimura
Department of Cardiovascular Medicine, Graduate School of Medicine, The University of Tokyo

Multi-color imaging

3D Reconstruction image of intact mouse adipose tissues
Blue / adipocytes:  BODIPY 
Green / Nuclei:  Hoechst33342
Red / collagen:  isolectin Gs-IB4
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Multi-color imaging

3D Reconstruction image of a blood vessel in mouse adipose tissues.
Blue / adipocytes:  BODIPY 
Green / Nuclei:  Hoechst33342
Red / endothelial cells:  isolectin Gs-IB4
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Escape of intracellular vesicles induced by mechanical stress

(By courtesy of Dr. Satoshi Nishimura, Department of Cardiovascular Medicine, Graduate School of Medicine, The University of Tokyo and Dr. Seiryo Sugiura, Department of Human and Engineered Environmental Studies, Graduate School of Frontier Sciences, The University of Tokyo.)

Multi-color imaging

Cultured NIH3T3 cells, excerpt from time-lapse images taken at 3.6 sec. interval.
Blue / Nuclei:  Hoechst33342 
Green / eGFP-Actin 
Red / Vesicles:  Cholera Toxin
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3D reconstruction image of blood vessels in mouse adipose tissues (unfixed)

(By courtesy of Dr. Satoshi Nishimura, Department of Cardiovascular Medicine, Graduate School of Medicine, The University of Tokyo and Dr. Seiryo Sugiura, Department of Human and Engineered Environmental Studies, Graduate School of Frontier Sciences, The University of Tokyo.)

Multi-color imaging

Blue / Nuclei:  Hoechst33342
Green / actin rhodamine
Red / eGFP-GULT4


Long-term,multi-dimensional livecell imaging

Long-term,multi-dimensional livecell imaging Long-term,multi-dimensional livecell imaging

Long-term, Six-dimensional Live-cell Imaging

Kazuo Yamagata, PhD, Wakayama Lab.(Laboratory for Genomic Reprogramming), Center for Developmental Biology, RIKEN
(Present post: Research Institute for Microbial Diseases, Osaka University)

IVF embryos were injected with a mixture of mRNAs for EGFP-α -tublin(Tublin,green) and H2B-mRFP1(H2B,red).The embryos were imaged by being exposed to two different wavelength of excitation(488 and 561 nm) at 7.5min intervals for about 70h and 51 images were acqired in the Z axis at each time point( total of 56,814 fluorescent images). As result of transplanting the embryos after imaging , all the pups were healthy, reproductively normal and not transgenic. Thus, this live-cell imaging technology is safe for full-term mouse development.

IVF:  in vitro fretilization

Long-term, Six-dimensional Live-cell Imaging

Left:  DIC
Right:  Confocal
Tubulin:  Green  
(EGFP- α -tubulin)
Null:  Red
(H2B-mRFP1)
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Micro PIV

Micro PIV

Visualization of warped interface between two fluids at a curve down stream of Y junction

(By courtesy of Seika Corporation (Japanese content).​ This experiment was done in the collaboration of the Oshima Laboratory, Institute of Industrial Science, The University of Tokyo.)

micro piv

Curved micro channel 
(width 100um×depth 50um)

micro piv

Shape analysis 
by volume rendering.

Instantaneous velocity distribution analysis of fluid flow in a round shape micro channel (width 100um×depth 50um)

(By courtesy of Seika Corporation (Japanese content). This experiment was done in the collaboration of the Oshima Laboratory, Institute of Industrial Science, The University of Tokyo.)

micro piv

Curved micro channel 
(width 100um×depth 50um)

micro piv

Shape analysis by volume rendering.

Related Products & Solutions

CSU-X1

Faster, brighter, and more versatility, the CSU-X1 is the advanced model of our CSU-series, which are widely recognized as the most powerful tools for live cell imaging.

Life Science

Your microscope can be easily upgraded to confocal microscope with Confocal Scanner Unit. The multi-beam scanning method offers not only high-speed imaging but also significantly reduced photo-toxicity and photo bleaching because of very reduced laser power of each beamlet. The CSU series have been already delivered more than 2,000 units and supported leading-edge research around the world.

 

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