In Vivo Trapping of Latex Bead Phagosomes for Quantitative Force Measurements

Methods Mol Biol. 2023:2623:187-200. doi: 10.1007/978-1-0716-2958-1_12.

Abstract

Optical trapping of organelles inside cells is a powerful technique for directly measuring the forces generated by motor proteins when they are transporting the organelle in the form of a "cargo". Such experiments provide an understanding of how multiple motors (similar or dissimilar) function in their endogenous environment. Here we describe the use of latex bead phagosomes ingested by macrophage cells as a model cargo for optical trap-based force measurements. A protocol for quantitative force measurements of microtubule-based motors (dynein and kinesins) inside macrophage cells is provided.

Keywords: Dynein; In vivo optical trapping; Kinesin; Latex bead phagosomes; Motors.

MeSH terms

  • Biological Transport
  • Dyneins / metabolism
  • Kinesins* / metabolism
  • Microspheres
  • Microtubules / metabolism
  • Phagosomes* / metabolism

Substances

  • Kinesins
  • Dyneins