Encyclopedia of Experiments: Immunology
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Transcript
Begin with a microfluidic cartridge equipped with storage tanks and unidirectional pumps that deliver reagents through channels into a reaction zone.
The reaction zone contains multiple spots immobilized with distinct allergens.
Load wash buffer, blocking buffer, peroxidase-conjugated secondary antibodies, and a chemiluminescent peroxidase substrate into the designated tanks.
Add a serum sample containing allergen-specific immunoglobulin E or IgE.
Initiate the analysis inside a chemiluminescence analyzer.
Within the reaction zone, the blocking buffer flows and blocks non-specific interaction sites.
The introduction of serum allows the IgEs to interact with the immobilized allergens.
Washing eliminates unbound IgEs.
Next, peroxidase-conjugated secondary antibodies flow in and interact with the immobilized IgEs.
Subsequent washing removes unbound antibodies.
The analyzer then introduces the chemiluminescent substrate and hydrogen peroxide .
The peroxidase oxidizes the substrate, emitting luminescence .
The luminescence intensity in each spot provides a quantifiable indication of distinct allergen-specific IgEs in the serum.
