Sequential Array Cytometry: Multi-Parameter Imaging with a Single Fluorescent Channel
| Autor: | Daniel R. Gossett, Noor S. Ahmed, Dino Di Carlo, Westbrook M. Weaver |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2010 |
| Předmět: |
Channel (digital image)
Microfluidics Population Biomedical Engineering Nanotechnology Cell Separation Biology Medical and Health Sciences Fluorescence Article Engineering Humans education Fluorescent Dyes Microlens education.field_of_study Single-cell Spectrometry business.industry Drug discovery Microfluidic Analytical Techniques Flow Cytometry Personalized medicine Spectrometry Fluorescence Hela Cells High-content screening Point-of-care High content screening Drug Caco-2 Cells Biological system business Cytometry discovery Immunocytochemistry Cellomics HeLa Cells |
| Zdroj: | Annals of Biomedical Engineering Annals of biomedical engineering, vol 39, iss 4 |
| ISSN: | 1521-6047 0090-6964 |
| Popis: | Heterogeneity within the human population and within diseased tissues necessitates a personalized medicine approach to diagnostics and the treatment of diseases. Functional assays at the single-cell level can contribute to uncovering heterogeneity and ultimately assist in improved treatment decisions based on the presence of outlier cells. We aim to develop a platform for high-throughput, single-cell-based assays using well-characterized hydrodynamic cell isolation arrays which allow for precise cell and fluid handling. Here, we demonstrate the ability to extract spatial and temporal information about several intracellular components using a single fluorescent channel, eliminating the problem of overlapping fluorescence emission spectra. Integrated with imaging technologies such as wide field-of-view lens-free fluorescent imaging, fiber-optic array scanning technology, and microlens arrays, use of a single fluorescent channel will reduce the cost of reagents and optical components. Specifically, we sequentially stain hydrodynamically trapped cells with three biochemical labels all sharing the same fluorescence excitation and emission spectrum. These markers allow us to analyze the amount of DNA, and compare nucleus-to-cytoplasm ratio, as well as glycosylation of surface proteins. By imaging cells in real-time we enable measurements of temporal localization of cellular components and intracellular reaction kinetics, the latter is used as a measurement of multi-drug resistance. Demonstrating the efficacy of this single-cell analysis platform is the first step in designing and implementing more complete assays, aimed toward improving diagnosis and personalized treatments to complex diseases. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|
Full text from SpringerLink