Researchers at the University Medical Center Göttingen and the University of Göttingen developed NanoFLex, a labeling method that visualizes eight different proteins in a single human cell simultaneously using standard microscopes. Published in ACS Nano, the technique distinguishes fluorescent labels by their fluorescence lifetime rather than color.
Overcoming the Color Barrier in Fluorescence Microscopy
Fluorescence microscopy has long served as a fundamental tool in biomedical research, allowing scientists to visualize specific proteins and cellular structures using antibodies carrying fluorescent dyes. When excited, the dyes fluoresce, and a microscope detects the light they emit. However, conventional microscopes can detect only three or four colors at a time—for example, blue/purple, green, yellow/orange and red. This color ceiling restricts researchers to viewing just a handful of proteins simultaneously in a single cell.
To break past this barrier, an international research team led by Dr. Felipe Opazo at the University Medical Center Göttingen (UMG) and Dr. Roman Tsukanov at the University of Göttingen introduced a second dimension to microscopy. According to the study published in ACS Nano, the team utilized fluorescence lifetime—defined in the reporting as the few billionths of a second a dye glows after being excited by laser light.
Two labels can look identical in color and still be cleanly separated, because one fluoresces measurably longer than the other. It is a bit
Dr. Felipe Opazo, University Medical Center Göttingen
How NanoFLex Codes Proteins
The mechanics behind NanoFLex—short for Nanobody-guided Fluorescence Lifetime multiplexing—rely on the immediate chemical environment of the fluorescent dyes.
Because these nanobodies are preassembled with the protein pockets before staining, laboratories can label all targets in a single step using conventional antibodies they already use routinely.
The method requires no specialized microscopy platform, functioning directly on standard microscopes, according to Dr. Roman Tsukanov.
Clinical Implications for Biopsies and Tissue Scans
The ability to read eight different proteins in immortalized human cells using a single staining step carries direct implications for clinical diagnostics.
Combined with the capacity to measure numerous markers from a single tissue section, this distinction makes the technique particularly attractive for analyzing biopsies where patient material is scarce and precious. As new fluorescent dyes and improved computational analysis methods become available, the study’s authors expect the number of simultaneously detectable proteins to rise further.