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Towards a spectrum-based bar code for identification of weakly fluorescent microparticles.
Zdeněk Petrášek1, Jens Wiedemann, Petra Schwille
1Max-Planck-Institut für Biochemie, Am Klopferspitz 18, D-82152 Martinsried, Germany. Biotechnologisches Zentrum, Technische Universität Dresden, Tatzberg 47/49, D-01307 Dresden, Germany.
Methods and Applications in Fluorescence
|November 18, 2017
Summary
This study demonstrates reliable identification of fluorescently labeled beads using spectrally resolved detection. The method accurately identifies spectral codes even with overlapping signals and strong background fluorescence.
Area of Science:
- Analytical Chemistry
- Spectroscopy
- Biotechnology
Background:
- Multiplexed detection of molecules is crucial for various applications.
- Fluorescent probes offer sensitive detection but identifying mixtures requires advanced methods.
- Spectral shape, noise, and detection channels impact identification accuracy.
Purpose of the Study:
- To investigate factors affecting spectral identification of fluorescent probes.
- To develop a method for estimating spectral identification performance.
- To demonstrate reliable identification of multiplexed fluorescent signals.
Main Methods:
- Utilizing spectrally resolved detection of fluorescent emission spectra.
- Analyzing the influence of spectral shape, experimental noise, and detection channels.
- Developing a formalism to estimate identification performance from reference spectra.
- Constructing a prism-based spectral detection setup.
Main Results:
- Seven distinct, overlapping spectral codes were reliably identified.
- Identification was successful even with autofluorescence stronger than the signal.
- A method was proposed to estimate spectral identification performance.
Conclusions:
- Spectrally resolved detection enables robust identification of multiplexed fluorescent probes.
- The developed method accurately identifies complex spectral codes in challenging conditions.
- This technique holds promise for analyzing unknown mixtures with fluorescent labels.
