Spectrally encoded microspheres for immunofluorescence analysis

Abstract

A modern immunofluorescence analysis based on spectrally encoded microspheres has found numerous and constantly growing applications in disease diagnosis, environmental supervision, and fundamental science. Here we present an overview of microsphere-based methods of multiplex immunofluorescence analysis and consider such important parameters of beads, that are crucial in most microsphere-based immunoassays, as size distribution, fluorescence stability, magnetic properties, and particle material. The preparation methods of the microspheres with tunable diameter, the introduction of various types of fluorochromes, and magnetic particles into the microspheres are discussed in details. This review also addresses the advantages and disadvantages of different approaches to implement technically bead-based immunofluorescence analysis. K E Y W O R D S bioengineering, biomedical applications, functionalization of polymers, synthesis and processing techniques 1 | INTRODUCTION Bead-based immunofluorescence analysis is a powerful tool widely used in numerous applications for life science research (extracellular vesicles [EV], 1 cytokines, 2 chemokines, 3 growth factors, cells and drug research, 4 etc.) and for in vitro diagnostics 5 of infectious diseases, diabetes, cardiovascular diseases, inflammation, cancer, 6

Local Sensing of Absolute Refractive Index During Protein‐Binding using Microlasers with Spectral Encoding

“…In turn, this enables the direct detection of antibody binding. In the future, the throughput can be improved further by forcing the particles into a stream passing through the laser beam one by one using microcapillary or acoustic focusing, [ 1 ] potentially allowing up to 1 kHz acquisition rates even with the equipment used in this study.…”

“…High-throughput, multiplexed screening of biomarkers is driving numerous advances in the fields of cellular biology, immunology, environmental testing, and drug research. One approach to DOI: 10.1002/adom.202300530 this is flow cytometry using polymer microspheres [1][2][3][4][5] or liquid droplets. [6][7][8] For example multiplexed detection is often achieved by using fluorescent microspheres with different emission colors, each encoded with a distinct capture antibody.…”

The influence of water on size of the monodisperse polystyrene microspheres prepared by dispersion polymerization

“…Micrometer-sized uniform polystyrene microspheres have a wide range of applications in biomedical engineering, biotechnology, electronics, and other fields. [1][2][3][4][5][6][7][8][9][10][11][12][13][14][15] For instance, these precisely sized and monodisperse microparticles can be effectively utilized in tandem with flow cytometry to craft fluorescently encoded microspheres. Moreover, they serve as templates for binding with magnetic nanoparticles, nucleic acid probes, and other biomolecular entities, thus finding application in the field of life sciences.…”

“…Moreover, they serve as templates for binding with magnetic nanoparticles, nucleic acid probes, and other biomolecular entities, thus finding application in the field of life sciences. [1][2][3][4][5][6][7][8][9] Traditionally, the preparation of monodisperse polystyrene particles ranging from 1 to 10 μm has involved various techniques, including suspension polymerization, [16][17][18] dispersion polymerization, [19][20][21] and seed polymerization. [22][23][24][25] Among these methods, suspension polymerization has proven to be inefficient in achieving monodispersity.…”

A single snapshot multiplex immunoassay platform utilizing dense test lines based on engineered beads

“…Herein, we propose an advanced platform that enables multiplex one-step immunoassays for timely on-site responses (MOnITOR) with robust and dense test lines based on engineered microbeads. Microbeads were incorporated because of their versatility (i.e., ease of surface functionalization, high stability of conjugates, high surface-to-volume ratio, and mobility), which can allow sensitive multiplex analysis via various encoding techniques ( Gilboa et al 2021 ; Kim et al 2017 ; Lee et al 2019 ; Norman et al 2020 ; Sankova et al 2020 ; Shrivastava et al 2020 ). By integrating versatile microbeads, target biomarkers can be easily changed, allowing adoption of this method during the ongoing global pandemic and to prepare for any potential co-epidemic threats and future pandemics ( Hemmig et al 2020 ).…”