Home › Cell Biology › In Vivo 4-Dimensional Tracking of Hematopoietic Stem and Progenitor Cells in Adult Mouse Calvarial Bone Marrow
Cell Biology JoVE (Open Access) Citable · DOI

In Vivo 4-Dimensional Tracking of Hematopoietic Stem and Progenitor Cells in Adult Mouse Calvarial Bone Marrow

DOI: 10.3791/51683-v
What you'll learn
  • ✓ Label and inject hematopoietic stem/progenitor cells for in vivo tracking
  • ✓ Prepare mouse calvarial bone marrow for two-photon and confocal microscopy
  • ✓ Acquire high-resolution 4D imaging data of HSPC interactions in bone marrow niche
  • ✓ Track HSPC movement and niche interactions over time using custom hardware setup
Protocol

The nature of the interactions between hematopoietic stem and progenitor cells (HSPCs) and bone marrow niches is poorly understood. Custom hardware modifications and a multi-step acquisition protocol allow the use of two-photon and confocal microscopy to image ex vivo labeled HSPCs homed within bone marrow areas, tracking interactions and movement.

Difficulty
advanced
Total time
~3–4 hours per mouse (including surgical prep, imaging, and post-imaging processing)
Model organism
Mouse (adult, calvarial bone marrow)
Biosafety
BSL-1

Steps

1
Label and inject hematopoietic stem progenitor cells

Ex vivo label isolated HSPCs with fluorescent markers and inject labeled cells into the mouse via appropriate route to enable homing to bone marrow.

▶ 01:52
2
Prepare mouse for in vivo imaging

Anesthetize mouse and surgically expose or position calvarial bone marrow region for microscopy access using custom hardware modifications.

▶ 03:21
3
Acquire high-resolution stacks and time-lapse data

Use two-photon and confocal microscopy to capture multi-stack high-resolution images and time-lapse sequences of labeled HSPCs within bone marrow tissue.

▶ 05:20
4
Analyze 4D imaging results and HSPC interactions

Process and analyze acquired 4D data to track HSPC movement, localization, and interactions with bone marrow niche components over time.

▶ 11:25
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