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    Micro-PET CT procedures for brain imaging of rats

    María Sanchiz Calvo, Eduard Bentea, christopher.cawthorne

    Published: 2024-02-29 DOI: 10.17504/protocols.io.e6nvwdw2zlmk/v1
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    Abstract

    Micro-PET CT procedures for brain imaging of rats

    Steps

    1.

    A range of PET radiotracers and radioligands (“radiopharmaceuticals”) can be used to

    image biochemical flux and receptor density in rodents. PET

    radiopharmaceuticals are administered in very low ‘tracer’ doses to avoid

    pharmacological effects, the specific activity of the tracer must be carefully

    evaluated to ensure this assumption is met.

    2.

    Typically rats are anaesthetised throughout the scanning process. Commonly this is with

    isoflurane (as described here), but other regimes may be necessary depending on

    the biological system to be studied. Whether animals are anaesthetised

    throughout the entire period of tracer uptake depends on both the relation of

    the tracer with the biological system being studied and the quantitation method

    used

    3.

    For studies of the rat brain, full length dynamic scanning allows a range of

    quantitative approaches to be compared. Typically rats are anesthetized in a

    warmed chamber before transfer to a warm mat for tail vein cannulation.

    Catheters are best constructed in-house using pharma-grade tubing of low

    diameter, to allow optimum length to be determined to allow injection when the

    animal is in the imaging cell in the PET scanner. Catheter needles are

    typically 23G or larger. Catheters should be checked for integrity immediately

    before scanning using sterile saline solution. Heparin may be added, though if

    the time between cannulation and injection is minimised this is optional

    4.

    The tail is cleaned with an ethanol wipe and warmed before a lateral vein is

    cannulated (this should be visible). In rats, the potency of the line can be

    confirmed by withdrawing a small amount of blood before reinjection. Cannulated

    animals must be placed in the specialised imaging cell before transfer to the

    scanner. Imaging cells differ between manufacturer, however temperature and

    respiration are usually monitored and this must be confirmed before scanning is

    implemented

    5.

    Syringe activities before injection should be measured in a dose calibrator, and

    residues after injection measured in the same way – importantly the exact time

    of measurement should be recorded. All equipment should be set to the same

    time.

    6.

    Typically the scan will be started before radiotracer is injected. Injection should be

    done over 20-30 seconds and the beginning and end of injection recorded, though

    if backpressure is felt this indicates the cannula is no longer potent and the

    injection should be abandoned. For this reason, syringe pump use requires

    additional validation. At the end of injection, the catheter should be removed

    if possible to allow measurement of residue. For quantitative scans, the

    catheters are not flushed.

    7.

    Dynamic PET images should be acquired for long enough to allow kinetic modelling. This

    depends on the radiotracer but times of  90-120 min are not uncommon. The brain should

    be at, or close to, the center of the field of view.

    8.

    After PET scanning, a CT image should be acquired for attenuation correction. The CT

    field of view must be equal or larger to the PET field of view.

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