Home › Cell Biology › Magnetic Stirrer Method for the Detection of Trichinella Larvae in Muscle Samples
Cell Biology JoVE (Open Access) Citable · DOI

Magnetic Stirrer Method for the Detection of Trichinella Larvae in Muscle Samples

DOI: 10.3791/55354-v
What you'll learn
  • ✓ Perform magnetic stirrer digestion of muscle samples for parasite detection
  • ✓ Execute filtration and sedimentation protocols for Trichinella larvae isolation
  • ✓ Conduct microscopic examination to identify and enumerate larvae
  • ✓ Apply the gold-standard method for trichinellosis prevention screening
Protocol

Biopharma Insights The prevention of human trichinellosis in many countries worldwide is based on the laboratory examination of muscle samples from susceptible animals by methods of digestion, of which the magnetic stirrer method is considered the gold standard.

Difficulty
intermediate
Total time
~4–6 hours per sample (digestion ~2 hrs, sedimentation overnight, microscopy ~30 min)
Biosafety
BSL-2

Steps

1
Prepare muscle samples for digestion

Weigh and prepare muscle tissue samples according to protocol specifications. This step establishes baseline material for subsequent enzymatic digestion.

▶ 00:44
2
Perform enzymatic digestion of muscle tissue

Subject prepared muscle samples to magnetic stirrer-assisted digestion using appropriate enzymes and buffer conditions. Maintain stirring throughout incubation to release Trichinella larvae from muscle tissue.

▶ 01:22
3
Filter digestion fluid to remove debris

Pass digestion fluid through filtration apparatus to separate undigested tissue and cellular debris from the suspension containing larvae. Retain filtrate for downstream analysis.

▶ 02:19
4
Measure and quantify undigested tissue residue

Determine the volume or mass of undigested tissue remaining after digestion to assess digestion completeness and ensure method validity.

▶ 03:26
5
Allow larvae to sediment in collection cylinder

Transfer filtrate to a sedimentation cylinder and permit Trichinella larvae to settle by gravity overnight or as specified. This concentrates larvae for microscopic observation.

▶ 04:50
6
Examine sediment under light microscopy

Pipette sediment aliquots onto microscope slides and perform systematic microscopic examination to identify and count Trichinella larvae morphology.

▶ 05:35
7
Document detection results and larvae quantification

Record microscopic findings, including presence/absence and enumeration of Trichinella larvae, to determine sample status and complete the assay.

▶ 07:39
💬 Comments coming soon