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Analytical Chem JoVE (Open Access) Citable · DOI

Manipulating Living Cells to Construct Stable 3D Cellular Assembly Without Artificial Scaffold

DOI: 10.3791/57815-v
What you'll learn
  • ✓ Use laser trapping to manipulate individual cells in three dimensions
  • ✓ Construct stable 3D cellular assemblies without artificial scaffolds
  • ✓ Apply dextran preparation and microscopy techniques for cell assembly visualization
Protocol

Biopharma Insights We demonstrate a novel method for constructing a single-cell-based 3-dimensional (3D) assembly without an artificial scaffold.

Difficulty
advanced
Total time
~2–4 hours per assembly (cell preparation through stable structure formation)
Model organism
Cell culture (specific cell type not specified in abstract)
Biosafety
BSL-1

Steps

1
Prepare and culture cells for manipulation

Culture and prepare cells to appropriate density and viability for laser trapping experiments. Cells must be in optimal condition to withstand manipulation without damage.

▶ 00:48
2
Prepare dextran solution for cell assembly

Prepare dextran medium according to protocol specifications. This medium facilitates cell-to-cell interactions and stabilizes the 3D assembly structure.

▶ 02:41
3
Configure laser and microscopy imaging system

Align and calibrate the laser trapping system and microscope setup for real-time visualization and manipulation of cells during assembly construction.

▶ 03:18
4
Manipulate cells using optical laser trapping

Use the laser trapping system to capture and position individual cells in three-dimensional space. Position each cell with precision to build the desired 3D structure.

▶ 03:55
5
Build three-dimensional cellular structure

Progressively assemble cells into a stable 3D configuration by continued laser manipulation and positioning until the desired structure is complete.

▶ 05:30
6
Analyze stability of assembled 3D structure

Acquire microscopy images and perform analysis to confirm structural stability, cell viability, and integrity of the scaffold-free 3D assembly over time.

▶ 06:10
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