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| 2D Image |
Reconstruction |
Segmentation |
Map Location |
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Project Information
Microscopy product ID: 7759
Image basename: Nf1_CKO
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Project (P2054)
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| Project ID |
P2054
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| Project name |
Characterization of optic gliomas from neurofibromatosis-1 (Nf1) genetically-engineered mice |
| Project description |
Use of large scale imaging of the optic nerve and chiasm to characterize development of optic gliomas from neurofibromatosis-1 (Nf1) genetically-engineered mice |
| Leader |
Keun-Young Kim
(Other projects)
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| Funding agency |
NCI and NCRR |
| Project start date |
2009-09-01 |
| Project end date |
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| Collaborators |
David Gutmann, Eric Bushong, Mark Ellisman, Da-Yong Lee |
| Publications |
Optic nerve dysfunction in a mouse model of neurofibromatosis-1 optic glioma.
Hegedus B, Hughes FW, Garbow JR, Gianino S, Banerjee D, Kim KY, Ellisman MH, Brantley MA Jr, Gutmann DH.
J Neuropathol Exp Neurol. 2009 May;68(5):542-51.
Ultrastructural characterization of the optic pathway in a mouse model of neurofibromatosis-1 optic glioma.
Kim KY, Ju WK, Hegedus B, Gutmann DH, Ellisman MH.
Neuroscience. 2010 Sep 29;170(1):178-88. Epub 2010 Jun 23. PMID: 20600672
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Experiment (7756)
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| Experiment title |
Large scale mosaic imaging with retrograde labeling of retinal ganglion cells
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| Experiment purpose |
The purpose of this study was to investigate the progression of changes in retinal ganglion cells and optic nerve glia in neurofibromatosis-1 (NF1) genetically-engineered mice with optic glioma. RGC were counted in whole retina preparations using high-resolution, mosaic confocal microscopy following their delineation by retrograde FluoroGold labeling. Then, we found reduced RGC numbers in Nf1+/¿GFAPCKO mice, supporting a model in which the combination of optic nerve Nf1 heterozygosity and glial cell Nf1 loss results in disrupted axonal-glial relationships, subsequently culminating in the degeneration of optic nerve axons and loss of their parent RGC neurons. |
| Experiment date |
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| Experimenter |
Keun-Young Kim |
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Subject group (7727)
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| Subject group name |
Retrograde tracing |
| Group by |
Injection of tracer
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| Treatment |
Both mutant and wild-type mice were injected with fluorogold tracer bilaterally into the superior colliculi. Please refer to the subject description for which images belong to which type of animal.
Retrograde Labeling of Retinal Ganglion Cells
One week prior to euthanasia, FluoroGold (1 ul/injection; Fluorochrome Inc., Englewood, CO) diluted in saline was microinjected bilaterally into the superior colliculi of anesthetized mice with a mixture of ketamine (Fort Dodge Animal Health) and xylazine (Vedeco Inc.) in a stereotactic apparatus, as previously described (Kim et al., 2004). Fluoro-Gold is taken up by the axon terminals of RGC neurons and transported retrogradely to the cell bodies in the retina (Selles-Navarro et al., 1996). The FluoroGold in the RGC neurons persists for at least 3 weeks without significant fading or leakage (Dong et al., 1996). Mosaic image arrays from retinal flat-mounts (n=6 retinal flat-mounts/mice/group) were captured under an Olympus spinning disk confocal microscope (Olympus America Inc., Center Valley, PA) equipped with a high-precision closed loop XY stage and closed loop Z control with commercial mosaic acquisition software from MicroBrightField (MBF Bioscience Inc., Williston, VT) modified by us (Chow et al., 2006, Price et al., 2006). The microscope was equipped with a high-resolution CCD camera for high-speed mosaic acquisition. Images were stored in Photoshop files (Adobe Systems Inc., San Jose, CA). |
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Subject (8794)
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| Species |
mouse |
| Scientific name |
Mus musculus
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| Strain |
C57Bl/6 Nf1 GFAP CKO |
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Organism (7756)
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| Age class |
Adult |
| Age |
9.0 months
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| Sex |
unspecified |
| Weight |
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| Vendor |
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| Cell Culture |
| Culture type |
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| Date plated |
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| Plating density |
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| Number of passages |
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| Media |
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| Cultured by |
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| Preparation |
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| Cell type |
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| Source |
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| Tissue product storage |
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| Cell line ID |
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| Lot number |
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| Notes |
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Tissue section or block (7757)
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| Anatomical location |
eye |
| Tissue product storage |
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| Microtome |
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| Thickness |
160.0 um
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| Orientation |
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Specimen Preparation Information
Microscopy product ID: 7759
Image basename: Nf1_CKO
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Protocol Details
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Protocol ID:
27696
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Protocol name:
Fluorogold injection
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Protocol descriptions:
Retrograde tracing with fluorogold
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Preparation steps:
- Staining
- Chemical Fixation
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| Stain (27694) |
| Stain name |
Fluorogold in saline
more details
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| Temperature |
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| Prepared by |
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| Notes |
Injected into superior colliculus |
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| Chemical Fixation (27695) |
| Fixative agent |
4% paraformaldehyde-0.1M phosphate buffer
more details
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| Time of fixation |
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| Vehicle |
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| Temperature |
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| Fixative volume |
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| Fixation method |
perfusion |
| Notes |
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Microscopy Product Information
Microscopy product ID: 7759
Image basename: Nf1_CKO
Microscopy product
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| Microscopy product id |
7759 |
| Image basename |
Nf1_CKO |
| Instrument |
Olympus DSU Confocal Microscope
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| X Size |
0
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| Y Size |
0
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| Z Size |
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| X Resolution |
0.39 nm/pixels
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| Y Resolution |
0.39 nm/pixels
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| Z Resolution |
1.0 nm/pixels
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| Create date |
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| PURL |
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| Product type |
MOSAIC
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| Microscope type |
Spinning Disc Confocal Microscopy
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| Session name |
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| Grid box count |
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| Grid info. |
N/A
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| Slide info. |
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| Plane count |
0
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| SRB ID |
0
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Imaging Product Types
Microscopy product ID: 7759
Image basename: Nf1_CKO
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Mosaic
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| Mosaic name |
Nf1_CKO
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| Mosaic description |
FG labled RGC in retinal flatmount |
| X position |
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| Y position |
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Specimen Description
Microscopy product ID: 7759
Image basename: Nf1_CKO
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Specimen description (7759, Nf1_CKO)
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| System |
central nervous system
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| Tissue |
retina |
| Organ |
eye |
| Region |
retina |
| Subregions |
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| Cell type |
retinal ganglion cell |
| Cell id |
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| Structure |
cell body |
| Atlas coordinates |
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| Atlas |
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| Map location |
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| Notes |
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Imaging Parameters
Microscopy product ID: 7759
Image basename: Nf1_CKO
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Light microscopy product (7759, Nf1_CKO)
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| Coverslip thickness |
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| Lens |
Olympus 20x dry |
| Lens magnification |
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| Numerical aperture |
.75 |
| Immersion medium |
air |
| Mounting medium |
gelvatol |
| Refractive index |
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LM channels
| Confocal channel (27725) |
| Filter used |
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| Fluorophor |
Fluorogold |
| Color |
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| Excitation wave length |
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| Emission wave length |
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| Laser type |
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| Notes |
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Specimen preparations
View complete protocol
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| Stain (27694) |
| Stain name |
Fluorogold in saline
more details
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| Temperature |
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| Prepared by |
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| Notes |
Injected into superior colliculus |
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2D Image Information
Microscopy product ID: 7759
Image basename: Nf1_CKO
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Downloadable data file description
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Full-resolution version
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N/A
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Down-sampled version
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N/A
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Raw data info
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Bit depth
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N/A
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Digitized by
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N/A
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| Digitizing platform |
N/A
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| Animation |
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No animation available.
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N/A
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| Data files |
| Action |
File type |
File size |
Description |
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Description: N/A
JSP Page
| Molecular target |
Chromagen |
Excitation λ |
Display color |
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Fluorogold in saline
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Fluorogold |
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No WIB image found.
Description: N/A
JSP Page
| Molecular target |
Chromagen |
Excitation λ |
Display color |
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Fluorogold in saline
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Fluorogold |
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Reconstruction Information
Microscopy product ID: 7759
Image basename: Nf1_CKO
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Reconstruction description
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Reconstruction description
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Full-resolution reconstruction file
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Assembled mosaic of a flat mount retina with retinal ganglion cells retrogradely labeled using fluorogold in tiff format
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Down-sampled version of the reconstruction file
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| Image map file |
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Reconstruction file format
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TIFF
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| Volume dimension |
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| Voxel scale |
(0.39, 0.39, 1.0) µm
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| Animation
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No animation available.
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| Data files |
| Action |
File type |
File size |
Description |
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Download
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512x512 image
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Unknown
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Mosaic reconstruction imaged using spinning disk confocal microscopy of a flat mount of retina showing retinal ganglion cells retrogradely labeled via fluorogold injection into the superior colliculus in an astrocyte specific Nf1 conditional knock out mouse. Details may be found in Kim et al., Neuroscience. 2010 Sep 29;170(1):178-88.
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Download
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Reconstruction data file
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214.6MB
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Assembled mosaic of a flat mount retina with retinal ganglion cells retrogradely labeled using fluorogold in tiff format
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Description: Mosaic reconstruction imaged using spinning disk confocal microscopy of a flat mount of retina showing retinal ganglion cells retrogradely labeled via fluorogold injection into the superior colliculus in an astrocyte specific Nf1 conditional knock out mouse. Details may be found in Kim et al., Neuroscience. 2010 Sep 29;170(1):178-88.
JSP Page
| Molecular target |
Chromagen |
Excitation λ |
Display color |
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Fluorogold in saline
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Fluorogold |
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Description: Mosaic reconstruction imaged using spinning disk confocal microscopy of a flat mount of retina showing retinal ganglion cells retrogradely labeled via fluorogold injection into the superior colliculus in an astrocyte specific Nf1 conditional knock out mouse. Details may be found in Kim et al., Neuroscience. 2010 Sep 29;170(1):178-88.
JSP Page
| Molecular target |
Chromagen |
Excitation λ |
Display color |
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Fluorogold in saline
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Fluorogold |
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Segmentation Information
Microscopy product ID: 7759
Image basename: Nf1_CKO
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| Message |
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No segmentation data is available for this data set yet.
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