Does your lab coat fit to your assay?

Area of Science:

• Biochemistry
• Analytical Chemistry
• Biotechnology

Background:

• Fluorescence-based assays are crucial for various scientific disciplines.
• Random spikes (artifacts) can compromise assay accuracy and data interpretation.
• Shorter-wavelength readouts, common in many assays, may be particularly susceptible to interference.

Purpose of the Study:

• To identify the cause of random spikes in fluorescence microplate assays.
• To investigate the role of lab coat materials in generating assay artifacts.
• To propose and validate a solution for reducing these unwanted signals.

Main Methods:

• Analysis of fluorescence properties of materials commonly found in laboratory environments.
• Comparison of fluorescence spectra of lint from cotton lab coats with known fluorescent compounds.
• Implementation of alternative lab coat materials in a high-throughput screening (HTS) campaign.
• Quantification of spike reduction after implementing the alternative lab coats.

Main Results:

• Lint from standard white cotton lab coats exhibits fluorescence properties similar to optical brighteners.
• These lint particles are identified as the primary source of randomly occurring spikes at 380 nm excitation and 450 nm emission.
• Utilizing non-cotton-based lab coats reduced assay spikes by 90% in an HTS setting.

Conclusions:

• Cotton lab coat lint is a significant, previously underestimated source of artifacts in fluorescence assays.
• The fluorescence of optical brighteners in cotton fibers contributes to signal interference.
• Adopting alternative, non-fluorescent lab coat materials is a practical and effective strategy to enhance assay data quality.