Home › Microscopy & Imaging › A Novel High-resolution In vivo Imaging Technique to Study the Dynamic Response of Intracranial Structures to Tumor Growth and Therapeutics
Microscopy & Imaging JoVE (Open Access) Citable · DOI

A Novel High-resolution In vivo Imaging Technique to Study the Dynamic Response of Intracranial Structures to Tumor Growth and Therapeutics

DOI: 10.3791/50363-v
What you'll learn
  • ✓ Perform bone marrow reconstitution and intracranial window implantation in mice
  • ✓ Apply stereotactic radiation to induce intracranial tumors
  • ✓ Acquire high-resolution two-photon microscopy images of brain tissue dynamics
  • ✓ Interpret single-cell vascular and parenchymal responses to tumor growth
Protocol

We describe a novel in vivo imaging technique that couples fluorescent chimeric mice with intracranial windows and high-resolution 2-photon microscopy. This imaging platform aids studies of dynamic changes in brain tissue and microvasculature, at a single-cell level, following pathological insults and is adaptable to assess intracranial drug delivery and distribution.

Difficulty
advanced
Total time
~4–6 weeks per mouse (bone marrow reconstitution 4 weeks, window surgery + recovery 1–2 weeks, imaging sessions ongoing)
Model organism
Mouse (fluorescent chimeric transgenic)
Biosafety
BSL-2

Steps

1
Perform bone marrow reconstitution in mice

Reconstitute bone marrow in chimeric mice to establish fluorescent cell labeling. This step typically requires myeloablation and transplantation, requiring ~4 weeks for engraftment and stabilization before subsequent procedures.

▶ 02:14
2
Implant intracranial imaging window

Surgically create and implant an intracranial window over the brain region of interest to enable optical access for two-photon microscopy. The window allows repeated longitudinal imaging while minimizing tissue damage.

▶ 03:34
3
Deliver stereotactic radiation to induce tumor

Use stereotactic radiation targeting to focally irradiate the intracranial region and induce tumor growth. Precise positioning via stereotactic frame ensures reproducible tumor initiation at the imaging site.

▶ 06:50
4
Acquire two-photon microscopy images in vivo

Perform high-resolution two-photon laser scanning microscopy on anesthetized mice to visualize fluorescently-labeled brain structures, microvasculature, and cellular dynamics at single-cell resolution over time.

▶ 08:39
5
Interpret imaging results and troubleshoot

Analyze dynamic changes in brain tissue and vasculature in response to tumor growth and therapeutics; address common technical challenges such as window clarity, phototoxicity, and motion artifacts.

▶ 09:53
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