Home › Neuroscience › Longitudinal Intravital Imaging of Brain Tumor Cell Behavior in Response to an Invasive Surgical Biopsy
Neuroscience JoVE (Open Access) Citable · DOI

Longitudinal Intravital Imaging of Brain Tumor Cell Behavior in Response to an Invasive Surgical Biopsy

DOI: 10.3791/59278-v
What you'll learn
  • ✓ Perform longitudinal multiphoton intravital imaging of brain tumor cells in living mice
  • ✓ Execute surgical biopsy procedures while maintaining imaging capability and animal viability
  • ✓ Analyze single-cell tumor behavior (migration, invasion, proliferation) before and after surgical intervention
Protocol

Here we describe a method for high-resolution time-lapse multiphoton imaging of brain tumor cells before and after invasive surgical intervention (e.g., biopsy) within the same living animal. This method allows studying the impact of these invasive surgical procedures on tumor cells' migratory, invasive, and proliferative behavior at a single cell level.

Difficulty
advanced
Total time
~4–6 hours per mouse (surgical prep, tumor establishment, imaging, biopsy, post-operative monitoring)
Model organism
Mouse (orthotopic brain tumor model)
Biosafety
BSL-1

Steps

1
Prepare surgical field and anesthetize mouse

Set up sterile surgical environment, anesthetize the mouse, and position for cranial access. Establish baseline physiological monitoring.

▶ 00:37
2
Inject tumor cells and prepare cranial imaging window

Stereotactically inject brain tumor cells into the mouse brain, then prepare and secure a cranial imaging window (CIW) for optical access to the tumor region.

▶ 02:16
3
Configure multiphoton microscope and optical alignment

Mount the anesthetized mouse on the imaging stage, align the objective lens with the cranial window, and calibrate laser power and detector gain for high-resolution multiphoton imaging.

▶ 03:26
4
Acquire time-lapse image sequences of tumor cells

Perform baseline time-lapse multiphoton imaging to capture tumor cell morphology, migration, and proliferation behavior before surgical intervention.

▶ 04:07
5
Perform biopsy-like injury and window reinsertion

Execute a controlled invasive surgical biopsy procedure (e.g., needle aspiration or micro-punch) adjacent to the tumor, then replace and reseal the cranial imaging window for post-injury imaging.

▶ 05:02
6
Process and analyze intravital imaging data

Extract and quantify single-cell trajectories, migration distances, velocity, invasion depth, and proliferation rates from pre- and post-biopsy image stacks using image analysis software.

▶ 05:50
7
Evaluate biopsy-induced changes in tumor cell behavior

Compare cellular dynamics before and after surgical injury to demonstrate alterations in migration, invasiveness, and proliferation in response to biopsy-like trauma.

▶ 06:56
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