SMALL ANIMAL IMAGING SYSTEM OV110

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1 SMALL ANIMAL IMAGING SYSTEM OV110 Olympus in vivo fluorescence molecular imaging systems: enhanced performance for translational research.

Low magnification A high performance fluorescence imaging From individual cells to entire small animals 0.14 (63 47 mm) 0.56 (16 12 mm) Maximum field of view: 78.

A wide range of magnifications The OV110 covers a wide range of magnifications, from 0.14x to 16x.

Low magnification 0.14x/ 0.56x/ 0.89x High-speed, high-precision, and high-sensitivity fluorescence imaging Proprietary Olympus' optics ensure brighter images.

2 Low magnification A high performance fluorescence imaging From individual cells to entire small animals 0.14 (63 47 mm) 0.56 (16 12 mm) Maximum field of view: 78.3 mm diameter Surgically exposed mouse lymph node metastasis imaged over a range of magnifications. A wide range of magnifications The OV110 covers a wide range of magnifications, from 0.14x to 16x. A series of four optimized objectives, parcentered and parfocal, provides seamless imaging of the entire body down to the single cell level without disturbing the animal. Low magnification 0.14x/ 0.56x/ 0.89x High-speed, high-precision, and high-sensitivity fluorescence imaging Proprietary Olympus' optics ensure brighter images. During low-magnification observation, shorter exposure times enable real-time observation of molecular changes in intact organs and tissues. Moving to higher magnification, fluorescently labeled molecules can be observed at the level of the single cell. High magnification x zoom CCD camera Emission filters Excitation filters zoom Motorized stage 1

High magnification system that supports multiple applications 1.6 16 zoom (5.5 4.1 0.55 0.41 mm) 0.89 (9.9 7.

, (USA) Support for a variety of fluorochromes Wide range of image acquisition methods The OV110 supports a wide range of fluorochromes, including fluorescent proteins,

The system is also corrected out to the near IR range to accommodate reagents that are preferred for in vivo imaging due to their increased penetration and Ty p i c a l p

DiI, Cy3, AlexaFluor 568, AlexaFluor 546, Rhodamine, RFP The OV110 permits selection of the optimal image acquisition method for customized experiments.

three-dimensional specimen and projects them in extended focus images), time-lapse imaging (which images an object over time at chosen intervals), and video recording. Cy5.

Complementary images of the same region at different fields of view and resolution are facilitated by the optical zoom capability.

3 High magnification system that supports multiple applications zoom ( mm) 0.89 ( mm) Images courtesy of Kensuke Yamauchi, Katsuhiro Hayashi, Meng Yang, Robert M. Hoffman AntiCancer Inc., (USA) Support for a variety of fluorochromes Wide range of image acquisition methods The OV110 supports a wide range of fluorochromes, including fluorescent proteins, dyes and particles. The system is also corrected out to the near IR range to accommodate reagents that are preferred for in vivo imaging due to their increased penetration and Ty p i c a l p ro b e s diminished scatter FITC, GFP, AlexaFluor 488, Cy2 through intact tissue. DiI, Cy3, AlexaFluor 568, AlexaFluor 546, Rhodamine, RFP The OV110 permits selection of the optimal image acquisition method for customized experiments. These methods include multi-point imaging (which acquires images in multiple sites repeated over time), Z-stack imaging (which acquires multiple focal planes of a three-dimensional specimen and projects them in extended focus images), time-lapse imaging (which images an object over time at chosen intervals), and video recording. Cy5.5, AlexaFluor 680, AngioSense 680, ProSense 680 Simple operation Cy7, AlexaFluor 750, AngioSense 750, ProSense 750 Longitudinal studies of the same anatomical location Complementary images of the same region at different fields of view and resolution are facilitated by the optical zoom capability. Time serial studies of biological response over a long period can be performed using the same animal. 15hr 18hr 21hr Graphic user interface control of camera exposure, illumination settings, magnification, filter selection, stage movement in X, Y and Z allows exact recording of experimental settings. Additional manual control of stage movement and focusing is possible. 24hr 27hr 30hr Time series observation of cancer cells in surgically excised skin flap of mouse. Cells were observed to extravasate by cytoplasmic extension. Images courtesy of Kensuke Yamauchi, Meng Yang, Robert M.Hoffman, AntiCancer Inc. (USA) 2

OV110 software Experiment manager Easy design and recall of various experimental procedures and data saving/read-out

Sophisticated experimental protocols are easily reproduced from one animal to the next or from one time point to the

Analysis One-click automatic measurement of fluorescence signal area and intensity Bone imaged non-invasively using

and intensity of fluorescence are automatically measured just by drawing regions of interest (ROIs) in images.

4 OV110 software Experiment manager Easy design and recall of various experimental procedures and data saving/read-out Each experimental procedure is easily designed just by dragging and arranging the icons. Sophisticated experimental protocols are easily reproduced from one animal to the next or from one time point to the next. A database function automatically saves images along with acquisition conditions and experimental procedures. Analysis One-click automatic measurement of fluorescence signal area and intensity Bone imaged non-invasively using near IR imaging agent (OsteoSense TM 680, VisEn Medical) Image courtesy of Aki Hanyu, Takeshi Imamura, Olympus Bio Imaging Lab. in collaboration with The Cancer Institute of the Japanese Foundation for Cancer Research(JFCR) Measurements of area and intensity of fluorescence are automatically measured just by drawing regions of interest (ROIs) in images. Measurement data can easily be displayed in a table and can be converted and saved in a file format for use in a variety of spreadsheet programs. Unmixing Tissue auto-fluorescence can obscure the fluorescent signal by its spectral overlap with fluorochromes in the green to red range. The OV110 s standard spectral unmixing feature enables separation of auto-fluorescence from target fluorescence signals. *UNMIXING function separates auto-fluorescence in the vicinity of GFP wavelengths 3

Example Applications Real-time imaging of dual-color cancer cells in blood vessels The dynamics of cancer cells in vessels were observed on the

The HT1080 human fibrosarcoma cell line expressing H2B-GFP in the nucleus and RFP in the cytoplasm was injected into host animals and the movement

microvessel 100µm arrested cancer cell microvessel 50µm artery Experimental method (Skin Flap technique) An arc-shaped incision was made in the

HT-1080 cells expressing GFP and RFP were injected into the epigastric cranialis vein through a catheter.

Development of Real-time Subcellular Dynamic Multicolor Imaging of Cancer-Cell Trafficking in Live Mice with a Variable-Magnification Whole-Mouse

April 15, 2006 Epigastrica cranialis vein Arc-shape incision syringe catheter Image courtesy of Kensuke Yamauchi, Meng Yang, Robert M.

(USA) in vivo imaging of tumor neovasculature using NIR agents Increased angiogenesis surrounding the tumor was observed non-invasively using NIR

5 Example Applications Real-time imaging of dual-color cancer cells in blood vessels The dynamics of cancer cells in vessels were observed on the lining of a surgically excised skin flap. The HT1080 human fibrosarcoma cell line expressing H2B-GFP in the nucleus and RFP in the cytoplasm was injected into host animals and the movement of cells in blood vessels was monitored over time. microvessel 100µm arrested cancer cell microvessel 50µm artery Experimental method (Skin Flap technique) An arc-shaped incision was made in the abdominal skin of the mouse, and then the skin was spread flat and fixed. HT-1080 cells expressing GFP and RFP were injected into the epigastric cranialis vein through a catheter. Superficial vessels were observed immediately after injection. Reference 1. Kensuke Yamauchi et al. Development of Real-time Subcellular Dynamic Multicolor Imaging of Cancer-Cell Trafficking in Live Mice with a Variable-Magnification Whole-Mouse Imaging System. Cancer Res 2006;66:(8).April 15, 2006 Epigastrica cranialis vein Arc-shape incision syringe catheter Image courtesy of Kensuke Yamauchi, Meng Yang, Robert M.Hoffman, AntiCancer Inc. (USA) in vivo imaging of tumor neovasculature using NIR agents Increased angiogenesis surrounding the tumor was observed non-invasively using NIR (Near Infrared) agents (AngioSense-IVM680; VisEn Medical). Cells from the HT1080 human fibrosarcoma cell line were implanted on the flank of a female nude mouse 8 days before observation. The image on the left shows the vasculature of the whole mouse using the 0.14x Macro Objective. In the image on the right, the magnification was increased to 16x using the zoom objective to image the microvasculature. The images were acquired with a 12-bit monochrome camera. 0.14x 16x Reference 1. Alencar, H. et al. Novel multiwavelength microscopic scanner for mouse imaging. Neoplasia. 7, (2005) Image courtesy of Aki Hanyu, Takeshi Imamura, Olympus Bio Imaging Lab. in collaboration with The Cancer Institute of the Japanese Foundation for Cancer Research (JFCR) 4

OV110 specifications This system is not intended for use in clinical applications High magnification mode Low magnification mode (3 individual parcentered and parfocal lenses) Zoom 1 2 3

4 16 12 63 47 Standard observation channels Brightfield/fluorescence 4 wavelengths [GFP region, RFP region, Near-IR region (680 nm/750 nm)] Light source for fluorescence excitation Xe or Hg-Xe lamp

6 OV110 specifications This system is not intended for use in clinical applications High magnification mode Low magnification mode (3 individual parcentered and parfocal lenses) Zoom Magnification Optical magnification Observation Observation range [mm, for a 2/3 inch CCD] Standard observation channels Brightfield/fluorescence 4 wavelengths [GFP region, RFP region, Near-IR region (680 nm/750 nm)] Light source for fluorescence excitation Xe or Hg-Xe lamp housing (150 W) Emission filter 6-position, filter diameter 32 mm Excitation filter 8-position, filter diameter 25 mm Brightfield illumination Light guide Light source: 100W Halogen lamp Stage High-precision motorized XY stage (X=118 mm, Y=152 mm) <controlled with software or the joystick> Temperature-maintainable thermoplate (from room temperature to 50 C) included Focusing Stage focus (Z<75mm), controlled with software or the focus handle Camera DP71 (recommended, 8.8x6.6 mm chip size), ImagEM (C ; Hamamatsu, Japan) (recommended, 8.2 x8.2 mm chip size): or other C-mount cameras (ask us for details) Camera control Auto-adjustment of exposure time (Auto, S-FL Auto) *when using DP71 Image acquisition area can be set by designating an ROI Software Image acquisition Still images (single-frame, time-lapse, or Z-stack image acquisition) Video (video recording in AVI format) Automated XYZ stage movement: allows accurate reproduction of stage position Experiment Manager: Automatic save of image acquisition conditions together with experimental procedure Analysis Analysis Image analysis/quantification: Average intensity calculation, area measurement etc. Separation of auto-fluorescence via Unmixing System control PC-AT compatible/os: Windows XP Professional SP2 (English ver.)/memory: 2GB or more/cpu: Pentium4, 2GHz or better/hard disk capacity: 80GB or more Power consumption PC: 7.0A Body: 13.0A Dimensions Body: 550(W) 554(D) 1000(H)mm, Desk (with caster): 1500 (W) 700(D) 680(H)mm Weight Approx. 200kg OV110 dimensions 550 (unit: mm) Dimensions Body: 550(W) 554(D) 1000(H)mm Desk (with caster): 1500 (W) 700(D) 680(H)mm Weight Approx. 200kg Compact and easy to use anesthesia equipment for laboratory animals Optional vaporization-type anesthesia device (Isoflurane only) allows long-term, stable state observation of animals. Anesthesia equipment 1, ,500 Notice: This system has many potential applications. Certain of these applications and materials used with them may require licensing under patents held by third parties. For example, AntiCancer, Inc. ( < and Xenogen Corporation ( < each own several patents in the field of imaging cells genetically engineered to produce light-emitting compounds. Olympus has obtained a license from AntiCancer, such that non-profit entities purchasing this system are licensed under AntiCancer U.S. Patent Nos. 6,232,523, 6,649,159, 6,235,968, 6,251,384, and 6,759,038 for noncommercial uses. Olympus' sale or other transfer of this system does not convey any additional rights or licenses under third party patents. It is suggested that users of this system consult with counsel to determine whether licensing of additional third party patents is required. Notation: All eight fluorescence mouse images on front page are artistic cut-outs of other images obtained invasively. Olympus is a registered trademark of Olympus Corporation and Olympus America Inc. All other brand and product names are trademarks of their respective holders. Certified ISO by UKAS ENVIRONMENTAL MANAGEMENT 008 IISO14001 Certification OLYMPUS CORPORATION Ina-Plant OLYMPUS CORPORATION has obtained ISO14001 Specifications are subject to change without any obligation on the part of the manufacturer. This catalog is printed by enviromentally-friendly waterless printing system with soy ink. Printed in Japan R394SB-0506

High-sensitivity. optical molecular imaging and high-resolution digital X-ray. In-Vivo Imaging Systems

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