How to Control Background in Fluorescence Lateral Flow Assays
Fluorescence assays are sensitive to background and reading stability. High background or signal drift can originate from bead coupling, membranes, pads, backing cards, buffers, sample matrix or reader settings.
Labeling material
Particle size, surface group, coupling efficiency, blocking strategy and storage conditions can influence signal and background.
Strip structure
NC membrane, sample pad, conjugate pad, absorbent pad and backing card jointly influence release, migration, nonspecific adsorption and reading region stability.
Reader window
Excitation source, filters, delay window, integration time and algorithm settings can influence final results, especially in TRF systems.
FAQ
What should be checked first for high background?
Blank strips, negative samples, conjugate release, backing card background, membrane lot and buffer compatibility should be checked step by step.
Can changing beads alone solve background?
Not always. Background is often a system issue involving beads, materials, buffers and reader settings.
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