| You will need to have Adobe Acrobat on your system to download the specifications for the imaging modality. Click here to download Adobe Reader. |
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| – Information – |
Fluorescence is the property of some atoms and molecules to absorb light at a particular wavelength and to subsequently emit light of a longer wavelength (i.e., when blue light is shown onto a certain type of protein it will appear to glow green). The discovery of green fluorescent protein (GFP) in the early 1960s enabled investigators to fuse the fluorophore moiety to a wide variety of protein and enzyme targets, in order to monitor cellular processes in living systems. Since its discovery, many derivatives have been developed that emit light across the breadth of the visible spectrum. However, because it requires an external light source to visualize the target, the target’s depth must be very shallow so that enough light can reach the target and be emitted back without scatter.
Fluorescence light imaging is rapid, painless, and harmless to the animal. The typical experiment involves shining excitation light of the desired wavelength on the subject at a fluence rate of approximately 4-20 mW/cm 2. This fluence rate is about the same as a brightly lit room or outside on a sunny day. The emission light reflected from the subject is then filtered and imaged with a camera. Acquisition of an image takes a few seconds to a few minutes (see basic specifications), depending on the intensity of the light. The Maestro® system is unlike most fluorescent systems because it can perform spectral unmixing and multiplexing. This enables the user to ‘remove’ autofluorescence from an image and to perform imaging using multiple fluorescent signals simultaneously.
Using the maximum field of view, up to three whole mice can be imaged simultaneously, while another three can be anesthetized in an external induction chamber. Using this staggered system, large sets of animals can be processed in a relatively short period of time. Visualizing the growth of tumors in which fluorescent proteins such as GFP or red fluorescent protein (RFP) are stably expressed is a common use for the system.
For more detailed information about the CRI Maestro Multi-spectral fluorescence. |
| – Specifications – |
Hardware
Field of View (length x width): 3.4 cm X 2.5 cm to 10.16 cm X 7.62 cm (variable zoom)
Resolution: 25 to 75 μm (based on zoom lens position)
Fluence Rate: 4 to 20 mW/cm 2 (based upon light position)
Scan Time: 5 sec to 1 min
Reconstruction Time: 1 to 30 sec
Scans needed for 1 mouse (Nose to Rump): 1 scan (at farthest table position) to 3 scans (at nearest table position)
Maximum Whole Mice per Scan: 3
Maximum Mice per Scan: 4
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Software
Operating System: Windows XP, 2000, NT
Supported File Format: TIFF
Image Data Types: 32-bit floating-point and 32-bit RGB color
Average File Size: 5 Mb to 10 Mb
Possible Analysis Techniques: Image one, two, three or four fluorophores simultaneously in the presence of autofluorescence in a single animal. Measuring the area, mean, standard deviation, minimum, and maximum of a selection or entire image. |
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Nuance/Maestro® software is the image acquisition and analysis product from CRI. It provides easy-to-use control panels that set and display all imaging parameters, from exposure time to wavelength ranges. Once acquisition is complete, the analysis section can quickly and accurately extract desired fluorescent signal from the autofluorescent background.
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Filter Types:
Blue
Green
Yellow
Red
Deep Red
Special NIR
NIR |
Excitation Range:
445 to 490 nm
503 to 555 nm
575 to 605 nm
615 to 665 nm
671 to 705 nm
725 to 775 nm
710 to 760 nm |
Emission Range:
515 nm longpass
580 nm longpass
645 nm longpass
700 nm longpass
750 nm longpass
800 nm longpass
800 nm longpass |
Fluence Rates:
4.14 - 17.34 mW/cm^2
3.49 - 14.68 mW/cm^2
2.92 - 12.32 mW/cm^2
3.47 - 14.42 mW/cm^2
2.81 - 11.71 mW/cm^2
2.86 - 12.13 mW/cm^2
2.84 - 12.01 mW/cm^2
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| – Rates – |
CRI Maestro multi-spectral fluorescence
Academic users: $200.00 per hour; billing is in 1/2 hour increments.
Corporate users: $300.00 per hour; billing is in 1/2 hour increments.
24 hour notice is required for changing or cancelling an appointment, or you will be charged for the scheduled time. |
| – Sample protocols – |
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Autofluorescence Signal Removal with Maestro Software |
| – Checklist and forms – |
The BIDMC and Longwood SAIF are murine viral pathogen-free and endo/ectoparasite-free animal facilities. All incoming animals must be of the identical health status. Prior to acceptance and transport of rodents to the BIDMC and Longwood SAIF a recent (within 3 months) health report must be submitted and reviewed by the BIDMC veterinarian. All decisions regarding animal import are at the discretion of the BIDMC veterinarian, and are final.
Checklist
Import Request Form
Transport Request Form (For BIDMC SL3 ARF only; all others must use the Import Request Form)
Customer Survey
Boiler Plate Language for Animal Protocol Amendments
Boiler Plate Language for Grant Applications and Papers
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Animals transported from BIDMC SL3* ARF |
Animals transported from within BIDMC but not SL3* ARF |
Animals transported to the BIDMC from an OUTSIDE institution |
Long-term studies
(> 1 day) |
- Animals can move back and forth with approved BIDMC protocol/amendment
- Only Transport Form is needed |
- The long-term animal facility is now open at the Longwood SAIF
- An approved BIDMC protocol/
amendment, Import Form, and Health Report are needed to use the satellite facility |
- The long-term animal facility is now open at the Longwood SAIF
- An approved Institutional protocol/
amendment, Import Form, and Health Report are needed to use the satellite facility |
Short-term studies
(<1 day) |
- Animals can move back and forth with approved BIDMC protocol/amendment
- Only Transport Form is needed |
- Animals can be imported once (terminal experiment) with an approved BIDMC protocol/amendment
- Import Form and Health Report are needed |
- Animals can be imported once (terminal experiment) with an approved Institutional protocol/amendment
- Import Form and Health Report are needed |
| *SL3 – BIDMC Slosberg-Landay 3rd floor Animal Research Facility. |
Other Notes:
- Previously euthanized animals or tissue specimens can be imaged if they are placed in a sealed plastic bag prior to shipment and must remain in the plastic bag while imaged; they must then be placed in a second plastic bag prior to disposal without protocol/health forms (i.e., animals can be imaged post mortem).
- After an animal has been euthanized, it can be taken back to its originating institution by the client for necropsy/histology.
- For short-term studies, all animals must be euthanized after imaging. In rare instances and only with written approval by the BIDMC veterinarian, live animals can be returned to their originating institution if a certain procedure in the approved animal protocol can be performed only at the originating facility and its endpoint is euthanasia by the end of the day (e.g., reperfusion of animal with formaldehyde for histology).
- Currently there is no overnight housing of animals at the BIDMC Animal Research Facility (Slosberg-Landay 3) until the completion of the new satellite facility (expected in summer 2007). Animals will only be accepted into the satellite if they are free of rodent viruses, endoparasites, and ectoparasites as per a recent health report.
- Animals that will come into the new, long-term satellite animal facility cannot be moved back to the SL3 facility without 8 weeks of quarantine.
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