The heterochronic parabiosis model was established 2 weeks after surgery. FMT4000 (PerkinElmer, Boston, MA, USA) was used to confirm that the partners in the model had a common blood circulation. FMT is a noninvasive technique utilising fluorescence, characterised by molecular specificity and sensitivity, and is used for tissue imaging in live animals [15 (link)]. ProSense 750-fluorescence agents (PerkinElmer, Boston, MA, USA) become fluorescent upon activation in the presence of cathepsins (cathepsins B, L, S, and plasmin), but do not fluoresce without any activation. ProSense 750FAST was injected into the old mouse in the heterochronic parabiosis model via the tail vein. The young mouse was imaged using FMT4000 24 hours after injection.
Protocol detail
Fluorescence Imaging of Cathepsin Activation in Heterochronic Parabiosis
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Animals
Blood circulation
Cathepsins
Cathepsins b
Fluorescence
Fluorescence agents
Mouse
Plasmin
Sensitivity
Surgery
Tail
Tissue
Vein
Corresponding Organization :
Other organizations : Second Hospital of Shanxi Medical University, Shanxi Medical University, Brown University, Providence College
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Variable analysis
independent variables
- Heterochronic parabiosis model (established 2 weeks after surgery)
- Common blood circulation between partners in the heterochronic parabiosis model (confirmed using FMT4000)
- Activation of ProSense 750-fluorescence agents in the presence of cathepsins (cathepsins B, L, S, and plasmin) in the old mouse (measured by fluorescence in the young mouse)
- Time of ProSense 750FAST injection (into the old mouse via the tail vein)
- Time of imaging the young mouse using FMT4000 (24 hours after injection)
- Positive control: Not explicitly mentioned.
- Negative control: Not explicitly mentioned.
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