Home › Cell Biology › Preparation and Respirometric Assessment of Mitochondria Isolated from Skeletal Muscle Tissue Obtained by Percutaneous Needle Biopsy
Cell Biology JoVE (Open Access) Citable · DOI

Preparation and Respirometric Assessment of Mitochondria Isolated from Skeletal Muscle Tissue Obtained by Percutaneous Needle Biopsy

DOI: 10.3791/52350-v
What you'll learn
  • ✓ Perform percutaneous needle biopsy of vastus lateralis muscle
  • ✓ Isolate and purify mitochondria from skeletal muscle tissue
  • ✓ Conduct respirometric profiling using extracellular flux assays
  • ✓ Assess mitochondrial oxidative capacity from clinical samples
Protocol

Biopharma Insights Methods for biopsy of Vastus lateralis, preparation of purified mitochondria, and respirometric profiling are described. The use of small muscle volume makes this technique suitable for clinical research applications.

Difficulty
advanced
Total time
~3-4 hours per sample (biopsy through respirometric analysis)
Model organism
Human (clinical biopsy samples)
Biosafety
BSL-1

Steps

1
Perform percutaneous needle biopsy of vastus lateralis

Collect small volume of skeletal muscle tissue from vastus lateralis using needle biopsy technique. This minimally invasive approach is suitable for clinical research applications.

▶ 01:38
2
Isolate and wash mitochondria from muscle tissue

Prepare purified mitochondria from biopsied muscle through differential centrifugation and washing steps. Remove contaminants and obtain intact mitochondrial fraction for downstream analysis.

▶ 03:20
3
Perform extracellular flux respirometric assays

Conduct XF assays to profile mitochondrial oxygen consumption rates and bioenergetic parameters. Measure real-time respirometric data to assess mitochondrial oxidative function.

▶ 06:35
4
Analyze and interpret respirometric assessment results

Review and interpret respirometric profiles to evaluate mitochondrial respiration capacity, ATP production, and metabolic health from the isolated mitochondrial preparation.

▶ 08:32
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