Home › Cell Biology › Studying the Protein Quality Control System of D. discoideum Using Temperature-controlled Live Cell Imaging
Cell Biology JoVE (Open Access) Citable · DOI

Studying the Protein Quality Control System of D. discoideum Using Temperature-controlled Live Cell Imaging

DOI: 10.3791/54730-v
What you'll learn
  • ✓ Set up temperature-controlled live cell imaging for D. discoideum protein aggregation studies
  • ✓ Quantify protein misfolding and stress response dynamics using image analysis
  • ✓ Interpret heat-induced proteostasis changes in social amoebae model systems
Protocol

Biopharma Insights The social amoebae Dictyostelium discoideum has recently been established as a system to study protein misfolding and proteostasis. Here, we describe a new imaging-based methodology to study temperature-induced protein aggregation and the cellular stress response in D. discoideum.

Difficulty
intermediate
Total time
~3–4 hours per experiment (cell prep + imaging + analysis)
Model organism
Dictyostelium discoideum
Biosafety
BSL-1

Steps

1
Prepare D. discoideum cells for live imaging

Culture and prepare Dictyostelium discoideum cells according to standard protocols. Ensure cells are in appropriate growth medium and at adequate density for microscopy.

▶ 00:35
2
Acquire temperature-controlled live cell images

Mount cells on microscope slides and use temperature-controlled imaging hardware to capture time-lapse fluorescence microscopy data during heat stress conditions. Record protein aggregation dynamics in real time.

▶ 01:19
3
Analyze aggregation and stress response images

Process time-lapse image sequences using image analysis software to quantify protein aggregate formation, intensity, localization, and cellular stress marker dynamics over the experimental period.

▶ 02:53
4
Interpret prion-like protein behavior during heat stress

Evaluate kinetics and spatial patterns of aggregation-prone proteins in D. discoideum under thermal stress, relating observed dynamics to proteostasis mechanisms and cellular quality control activation.

▶ 04:05
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