phoenix micron iv retinal imaging microscope

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The Micron IV Retinal Imaging Microscope delivers precision in vivo imaging capability for mice and rats with stunning quality bright field images. Access to a range of fluoropores has been demonstrated including GFP, YFP, CFP, and mCherry. Fluorescein and Evan’s Blue angiograms can be obtained which show the flow of fluorescein-stained red blood cells in the capillaries.

The generation four Micron technology reflects Phoenix Research Labs’ continuing commitment to innovation. Incorporated in this advancement is a custom built three-chip CCD camera with several performance enhancements, improvements in the ergonomic design, and provision for four filter positions.

Improved sensitivity and color saturationA new technology three-chip CCD provides improved sensitivity and better signal processing. Imaging of yet fainter fluoropores and color saturation is greatly improved. Near infrared (NIR) imaging Through custom design is the capability to image in the near infrared, between 700 and 900 nm. This feature provides access to other fluoropores and potentially ICG angiograms. It is a key technical advancement for the auxiliary Phoenix Image-Guided Focal ERG system. Retinal features are visible at 850nm so that eye can be positioned to the area of interest while maintaining dark adaptation for scotopic testing.

Improved versatility and ergonomicsThe Micron IV provides imaging of both the mouse and rat retina through objective lenses optimized for each animal. The new system has additional ergonomic features which make an already easy to use research tool even more convenient.

In vivo validation Longitudinal research depending on retinal histology alone is a thing of the past. Complete a comprehensive in-vivo study in just minutes. Conveniently screen large numbers of animals, save costs and strengthen the quality of research through longitudinal imaging. Become empowered with the Micron IV to make new discoveries into retinal disease and development.

Phoenix Micron IV Retinal Imaging Microscope

Bright field images (left to right): rat bright field image, mouse fluorescein angiogram, mouse CFP, zebrafish bright field image, mouse with retinal degeneration, gerbil bright

Phoenix Micron IV Retinal Imaging Microscope with animal stage.

6920 Koll Center Pkwy, #219

Pleasanton, California 94566

925.485.1100 T 925.485.1155 F

www.phoenixreslabs.com

Eye To Eye With Precision

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Phoenix Micron IV fuels researchThe Phoenix Micron IV, designed specifically for laboratory animals, delivers a unique proposition for image-guided eye research. The platform supports a family of additional turnkey research instruments including Image-Guided Laser, Slit Lamp Imaging, Focal ERG and Image-Guided OCT. Unlike stand-alone adaptations of human instruments, these range of products interface directly with the Micron IV to support image-guided comprehensive studies without requiring use of additional limited laboratory bench space.

Powerful, validated, acceptedThe Phoenix Micron technology has contributed to over 125 research centers in Asia, North America and Europe and over 40 publications in print. With its broad range of applications including basic research, toxicology, pharmaceutical efficacy testing and neurological research, the Micron retinal imaging microscope is sure to continue to fuel scientific discoveries fulfilling the Phoenix mission to contribute to the war on blindness through technology innovations.

PHOENIX MICRON IV RETINAL IMAGING MICROSCOPE

SPECIFICATIONS

Retinal imaging resolution 4 μm or better (mouse) 8 μm (rat)

CCD pixel resolution 2 μm (mouse) 4 μm (rat)

CCD sensor Low noise 3 chip CCD with extended imaging in the near infared (NIR)

Resolution 1024x768 pixels

Depth of focus 20 μm

Range of focus Retinal surface to cyrstalline lens

Field of view 50 degrees; 1.8mm (mouse) 3.6mm (rat)

Signal to noise ratio 60db

Image format AVI, BMP, JPEG, PNG, Tiff

Imaging Modalities Bright Field 450-650 nm

NIR up to 900 nm

Fluorescein Angiography 469-35 nm

Fluorescent imaging of arbitrary fluoropores

Animal stage 3 degrees of rotation and 3 degrees of translation

Objective lenses Separate objective lenses for mouse and rat

Camera head stand Rack and pinion motion along axis, 2 axis rotation

Exposure time up to 100x long exposure

Filter wheels 2 wheels with 4 slots (excitation and emission)

Light source Xenon bulb

CPU Intel Core i7 3.1GHz w/ MS Windows 7 Professional and proprietary imaging software

DDR3 Ssamsung SDRAM 8GB

Monitor Wide screen LCD 22 inch

Accessories Mouse, Keyboard, Footswitch

Phoenix Micron IV Retinal Imaging lenses for mice and rats.

Designed for Small Animal Imaging The Phoenix Image-Guided OCT is optimized for eye research using laboratory animals such as mice, rats and zebrafish. The remarkably small stacking design is integrated into the Micron IV retinal imaging microscope. There is no further footprint space required over that of the base platform technology; the Micron IV.

See the whole picture, and moreVisualize the precise location of the OCT scan using the real-time Micron IV bright-field image. A superimposed line directly on the image placed over the retinal feature being examined delivers cohesive information about the sample.

Maximal clarityKeep the cornea and lens clear easily using a coupling gel interface between the specially designed rat or mouse objective lens and the eye. Lens contact also minimizes translational movements made by respirations of the animal which can blur the image.

The Phoenix OCT provides state of the art resolution of below 2 microns longitudinally. This combined with the Micron IV retinal imaging microscope delivers high resolution in three dimensions.

Fluorescein-guided OCTCombining the OCT scan with fluorescein is possible with the simple turn of a filter. Emission patterns guide the beam placement to the exact retinal loci. Utilize an enhanced depth imaging technique to view the choroid and even the sclera in some animals. Perform circle scans to evaluate retinal nerve fiber thickness and 3D volume scans for enface views.

Comprehensive imaging for comprehensive outcomes With the high level of integration and ease of use, in a single setting quickly obtain bright-field images, an angiogram, and an OCT scan. This avoids separate sedations which some animals cannot tolerate without a calendar time delay.

In vivo validationLongitudinal research depending only on histology is a thing of the past. Complete a comprehensive in-vivo retinal study of OCT and bright-field images with corresponding angiograms in just minutes. Document the layers in histologic detail by OCT

Phoenix Image-Guided OCTOptimized for Small Animal Imaging

Phoenix Image-Guided OCT system (top to bottom):Brown Norway rat, corneal scar on mouse, zebrafish retina, laser burns on a mouse.

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and correlate the time points of imaging with a tissue sample at the end point of the study. Beyond convenience, documenting morphology over time using using the Micron Image-Guided OCT is an essential tool for total understanding of retinal structure.

Phoenix Image-Guided OCT fuels researchThe Micron IV design delivers a unique proposition for image-guided eye research tailored specifically for laboratory animals. The Phoenix Micron IV platform supports a family of additional turnkey research instruments including Image-Guided

Laser, Slit Lamp Imaging, and Focal ERG. Unlike stand-alone adaptations of human instruments, the Phoenix range of products interface directly with the Micron IV to support image-guided comprehensive studies. With a broad range of applications including basic research, toxicology, pharmaceutical efficacy testing and neurological research, the Micron IV is sure to fuel scientific discoveries, which is at the heart of the Phoenix mission.

The small compact design of the Phoenix Image-Guided OCT engineered specifically for small animal research.

PHOENIX IMAGE-GUIDED OCT

SPECIFICATIONS

Methodology Spectral Domain OCT

Image guidance Bright-field live fundus image

Lightsource Ultra broadband (160nm) SLD centered at 830 nm

Transverse resolution 2 μm (mouse) 4 microns (rat)

Imaging depth 1.4 mm in tissue

Axial resolution 2μm in tissue

Imaging speed 10,000 - 20,000 A-scans per second

Pixels per A-scan 1024

Maximum output power 750 microwatts

File formats JPEG, PNG, TIF, VTI, Raw

Scan Patterns Line, Circle, 3D Volume

Measurement tools Software Calipers

Animal models Mouse, rat, zebrafish

Objective Lenses Separate Phoenix Micron IV standard objective lenses for mouse and rat

Separate Phoenix OCT objective lenses for mouse and rat

Integrated with the Phoenix Micron Retinal Imaging Microscope and associated hardware

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Compact, precise laser deliveryThe Phoenix Research Labs’ Image-Guided Laser System is user-friendly, producing precise, easy-to-deliver laser energy to facilitate research into the mechanisms of retinal damage and regeneration.

Designed for small animalsThe Image-Guided Laser System is coupled with the Micron IV Retinal Imaging Microscope, both of which are designed specifically for small animal research. No additional lab, bench space or clinical slit lamp delivery system is needed.

Easy TargetingLaser targeting using image-guidence is much easier and more precise due to the magnification of the bright field image. Simply align the animal to the Micron, position and focus the aiming beam, adjust the laser settings and deliver the energy.

Clear advantages With the cornea coupled to the objective lens using a gel interface, the eye is stabilized against movements from respirations and delivered power is more consistent than through open air. The coupling gel maintains hydration of the cornea, reducing the chances of media opacification.

Publication qualityUse the bright field image as a guide to place the aiming beam on target areas. Easily capture photographs or videos for publications or presentations to strengthen research documentation. Integration with the Phoenix Micron IV Retinal Imaging Microscope enables the recording of pre/post laser treatment spots as well as documentation of longitudinal studies.

Complete a comprehensive in-vivo retinal study at the spin of the filter wheel by performing fluorescein angiography to observe and document leakage patterns.

Phoenix Image-Guided Laser System

Image-Guided Laser System (top to bottom):Laser burns of varying intensities on a rat, fluorescein angiography on mouse with OCT beam overlay, corresponding OCT of penetration of laser burn on mouse.

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Phoenix Image-Guided Laser fuels your researchThe Micron IV design delivers a unique proposition for image-guided eye research tailored specifically for laboratory animals. The Phoenix Micron IV platform supports a family of additional turnkey research instruments including Slit Lamp Imaging, Focal ERG, and Image-Guided OCT. Unlike stand-alone adaptations of human instruments, the Phoenix range of products interface directly with the Micron IV to support your image-guided comprehensive studies. With a broad range of applications including basic research, toxicology, pharmaceutical efficacy testing and neurological research, the Micron IV is sure to fuel scientific discoveries, which is at the heart of the Phoenix mission.

The small, compact, easy to use Phoenix Image-Guided Laser makes research into retinal ablation simple.

PHOENIX IMAGE-GUIDED LASER SYSTEM

We have done our research so that you can do yours.

SPECIFICATIONS

Laser wavelength 532 nm

Spot size 50 μm

Aiming beam laser wavelength 635 nm

Pulse duration 0-5 seconds

Maximum delivered laser power 0-600 mW

Laser Source Meridian Merilas 532 green laser photocoagulator p/n 100470

Lenses Separate standard Micron IV objective lenses for mouse and rat

Separate laser objective lenses for mouse and rat


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It is the first of it’s kind. The Phoenix Anterior Segment Slit Lamp Imaging System, specifically designed for small animal imaging of mice and rats, is a full-featured ophthalmic slit lamp scaled for the size of laboratory subjects.

Compact, diverse utilityNo additional bench or laboratory space is required to achieve superior quality images with the integration of the Phoenix Slit Lamp Imaging System to the Micron IV Retinal Imaging Microscope. Image not only the posterior segment of the eye detail, but document the cornea and lens too.

The system provides bright field imaging as well as a cobalt blue filter for documenting corneal staining with fluorescein. This slit lamp offers variable slit width and intensity as well as dual back-fill lamps for added flexibility of illumination.

Diffuse illumination By using one or both of the two back fill lights, achieve overview pictures of the eye and adnexa.

Direct focal illuminationBy adjusting the slit beam width and/or height, the subject of interest can be illuminated directly at the plane of focus.

Indirect illumination Scattered reflected light from a slit beam positioned adjacent to the pathology will illuminate subtle corneal structures.

Retroillumination By bouncing light from the back of the eye or the iris, corneal or lens pathology can be illuminated from behind.

Phoenix Anterior Segment Slit Lamp Imaging

Retroillumination of penetrating keratoplasty (rat)

Diffuse illumination of keratitis (mouse)

Indirect illumination of keratitis (mouse) with fluorescein staining (right)

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Phoenix Slit Lamp imaging augments researchThe Phoenix Micron IV design delivers a unique proposition for image-guided eye research tailored specifically for laboratory animals. The platform supports a family of additional turnkey research instruments including Image-Guided Laser, Focal ERG and Image-Guided OCT. Unlike stand-alone adaptations of human instruments, the Phoenix range of products interface directly with the Micron IV to support image-guided comprehensive studies. With a broad range of applications including basic research, toxicology, pharmaceutical efficacy testing and neurological research, the Micron IV is sure to fuel scientific discoveries, which is at the heart of the Phoenix mission.

Phoenix Anterior Segment Slit Lamp Imaging is compact and tailored for small animal imaging.

PHOENIX ANTERIOR SEGMENT SLIT LAMP IMAGING SYSTEM

Document research every step of the way with the Phoenix Anterior Segment Slit Lamp Imaging System.

SPECIFICATIONS

Slit beam light source 5w LED

Back fill light source two 5w LEDs

Filter Blue exciter 463/35

Radial movement of slit column +/- 90° to axis

Slit lamp LED controller on/off switch for slit lamp back fill lights + intensity controls


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The Phoenix generation II Image-Guided Focal ERG is optimized for use with mice and rats. The unique design utilizes the new near infrared (NIR) imaging capability of the Micron IV to precisely select a location for focal testing. Testing of a selected specific region is valuable to differentiate segments of the retina as to functionality. Such a capability is ideal for applications for investigating retinal response where therapy is provided on a regional basis. The Generation II Phoenix Image-Guided Focal ERG attaches to the Micron IV and uses its imaging capability.

Scotopic testing with Generation II The Micron IV, the next generation retinal imaging microscope, features a custom three-chip CCD with improved sensitivity and capability for imaging at 850 nm. The rodent retinal response is lower by 10^-8 than at the peak in the green. This use of the 850 nm illumination allows guiding of the focal spot while also maintaining scotopic adaptation.

Controlling stimulus and receptionA low-noise microprocessor-based control system provides for illumination and reception through easy to use software screens. The LED light sources deliver aerial energy densities from well above the ISCEV high standard of 100 Cd sec/m^2 to well below the lower standard of 0.01 Cd sec/^2. A sophisticated software based system provides for illumination control over six orders of magnitude and pulse length control from 0.2 to 500 milliseconds, to separate cone from rod response, set backgrounds or deliver “flicker”.

Unique electrode system Contact lens electrodes can be challenging to use as they can easily disconnect from the tiny eyes of rodents. Additionally insertion must be accomplished in a dark laboratory if one is to conduct scotopic testing.

The Image-Guided Focal ERG design features a corneal electrode integrated into a gold-tipped ring at the end of the objective lens which couples with the small animal eye. Stimulus targeting is done under guidance from deep NIR illumination and an IR camera. With the animal eye coupled to the objective lens/electrode, the user knows with confidence that it will remain in place.

Phoenix Image-Guided Focal ERG

Phoenix Image-Guided Focal ERG stimulus is projected against a dim red fundus image for targeting (top left). Once the animal can be light adapted, corresponding bright field images can be captured (top right). Focal ERG traces from C57BL/6 mouse with increased brightness by a factor of two (bottom).

Phoenix Image-Guided Focal ERG nosepiece with integrated corneal electrode. As an attachment to the Micron IV the Image-Guided Focal ERG does not require additional laboratory bench space.

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Dark Lab Technology Absolute dark adaption is required for scotopic testing. The Image-Guided Focal ERG uses light at 850 nm to illuminate the retina for targeting. The laboratory is illuminated at 750 nm which the human eye can use for handling the equipment.

Animal HandlingThe animal is provided with a calibrated heating pad which mains body temperature to within 1 deg C.

Testing S-cones and M-cones The Phoenix Image-Guided Focal ERG provides for illumination at 360, 504 and 850 nm. This facilitates a unique capability to test the M and S cones. To access the rods, the system delivers the capability to establish a continuous background illumination and to provide flicker.

SPECIFICATIONSStimulation white light LED

focal spot size diameters 0.25mm, 0.5mm, 0.75mm, 1mm, 1.5mm in mouse (doubled for rat)

micrometer driven targeting

Additional stimulation 360 nm, 505 nm, 850 nm, others by request (in development)

Temporal control pulse length 0.2 to 500 milliseconds

pulse delay

flicker

continuous background

Range of intensity 10^6 (100 Cd sec/m^2 to 10-4 Cd sec/m^2)

Dark Lab technology illuminate the eye for alignment at 850 nm

laboratory illunination at 750 nm

red screen monitor cover 22”

Electrodes tail (ground), reference (gold-plated bite bar), corneal (gold-plate on objective lens)

Objective lenses separate Phoenix Micron IV standard objective lenses for mouse and rat

separate Phoenix Focal ERG objective lenses for mouse and rat

Software and controls for reception and analysis

averaging with user-selectable rejection of poor traces

controllable bandwidth

controllable digitization rate

controllable scan time

analysis of implicit time and peaks in scan

automatic generation of waterfall display

export into convenient formats

Heater mouse and rat



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The Phoenix Generation Two Image-Guided Focal ERG provides LED based illumination optimized to study different classes of photoreceptors.

The Phoenix Ganzfeld ERG takes advantage of the Maxwellian view illumination technique to avoid the large size of the traditional bowl illumination approach. It is uniquely and specifically designed as a stand alone Ganzfeld for the rodent eye with the ability to test the S-cones, M-cones and rods of these small animals.

Maxwellian view illumination The Maxwellian view illumination technique focuses the light from a single LED onto the plane of the eye’s iris using a small f/number lens diverging the light to illuminate the entire retina. This approach, with its compact size and use of LEDs, requires only 50 cm of lab bench space.

Illumination designed for the rodent eyeThe Phoenix Ganzfeld ERG is not an ad hoc adaptation of a clinical instrument designed for the human eye. While the rodent eye is a useful analogue to the human eye there are substantial differences. As shown in the side bar there are only two classes of cones in rodents; S-cones sensitive to ultraviolet (UV) light and M-cones sensitive to green light. In addition, the rods are also sensitive to green but have UV sensitivity as well. A green LED emitting at 510 nm is used to excite the M-cones and rods while the UV LED emitting at 360 nm is used to excite the S-cones. To separate the rod and the cone response, the system is equipped with flicker and background capabilities.

The use of a single LED light source not only preserves bench space but provides extraordinary flexibility in separately testing each of these photoreceptor classes.

A near infrared (IR) LED at 850 nm is used for alignment. This is a wavelength were there is no response from the rodent retina, thereby the very best possible levels of scotopic adaptation are maintained.

This complete set of testing modalities is unique to the Phoenix Ganzfeld and is driven by a convenient software menu. The system is completed by an outstanding set of analysis, display, and data extraction routines.

Phoenix Ganzfeld ERG

400 500 600 700

0

1

Wavelength (nm)

rod

M cone

S cone

UV excita*on Green excita*on

The Phoenix Ganzfeld uniquely studies the rodent M and S cones and rods using narrow band illumination in the green and UV wavelengths.

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LED

Light focuses at the pupil and fills the eye



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Configuration of the Maxwellian view Ganzfeld illumination The Maxwellian view Ganzfeld approach makes changing the light source is easy. The system can easily host flash lamps and other sources via a fiber optic. The base LEDs in the body can be changed giving the research flexibility and convenient access to various wavelengths.

Unique electrode systemThe Phoenix Ganzfeld ERG design features a unique corneal electrode design. It is integrated into a gold-tipped ring attached to the objective lens which couples with the small animal eye. Guidance using deep NIR illumination and an IR camera allows precise alignment of the eye to the light and electrode while maintaining scotopic adaptation.

SPECIFICATIONSLED wavelength aerial energy density from above 100 CD sec/m^2 to below 0.01 Cd sec/m^2.

Set levels over a range of 10^6

LED light sources 360 nm, 504 nm, 850 nm, others by request

Temporal control pulse length from 0.2 to 500 milliseconds

pulse delay

flicker

continuous background

Dark lab technology red monitor overlay 22”

iIlluminate the eye for alignment at 850 nm

laboratory illumination at 750 nm

Objective lenses separate Phoenix Ganzfeld ERG objective lenses for mouse and rat included

Software and controls for reception and analysis (can be used with both the Ganzfeld and the Phoenix Image Guided Focal ERG)

averaging with screen based rejection of poor traces

bandwidth

digitization rate

scan time

analysis of implicit time and peaks in scan

automatic generation of waterfall display

export into convenient formats

Electrodes tail (ground), reference (gold-plated bite bar), corneal (gold-plate on objective lens)

Heater mouse and rat


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Green and UV traces under different levels of energy, pulse length and dark adaptation illustrate the flexibility of the Phoenix Ganzfeld system to test S and M cones. To separate the rod and cone response, the system is configured to take advantage of the rod saturation by providing continuous background or by flickering the stimulus at a rate higher than the rods can respond.

Left side:Both traces use 510 nm green light. The animal in the upper trace is photoptic and the lower trace is scotopic. Saturation of the B and A wave has been seen in various recordings,

Right side:These traces use 306 nm UV light. The upper trace is scoptoic and a limited A wave is shown. The lower trace is photoptic and a strong oscillatory potential (OP) signal is seen along with signs of B wave saturation.

ERG Ganzfeld recordings in UV and Green Light

phoenix micron iv retinal imaging microscope

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