Cell-Based No-Wash Fluorescence Assays for Compound Screens Using a Fluorescence Cytometry Plate Reader

Area of Science:

• Biotechnology
• Cell Biology
• Assay Development

Background:

• High-throughput cell-based fluorescent imaging assays often require background signal removal, which is challenging due to equipment needs and variations from multiple wash steps.
• Traditional methods involve extensive plate processing, leading to increased time and data file sizes.

Purpose of the Study:

• To develop a wash-free cell-based fluorescence assay for high-throughput screening.
• To improve assay efficiency, sensitivity, and data quality while reducing artifacts and processing time.

Main Methods:

• Development of a wash-free cell-based fluorescence assay using a laser scanning fluorescence plate cytometer.
• Application of the assay to Niemann-Pick disease type A and C, and detection of Zika and Dengue viral envelope proteins using the Mirrorball method.

Main Results:

• The wash-free assay demonstrated robust screening results for lysosomal storage diseases.
• The method was successfully applied to cholesterol detection in Niemann-Pick disease type C.
• Preliminary results for viral envelope protein detection were obtained using the Mirrorball method.

Conclusions:

• The Mirrorball-based method significantly improves throughput and data quality in image-based screening.
• This versatile assay format reduces assay artifacts, increases sensitivity and efficiency, and requires less researcher intervention.