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Correlative Light- and Electron Microscopy Using Quantum Dot Nanoparticles
Published on: August 7, 2016
Time-resolved fluorescence imaging for quantitative histochemistry using lanthanide chelates in nanoparticles and
1Department of Urology, Lund University, Malmö University Hospital, SE 205 02 Malmö, Sweden. ville.vaisanen@utu.fi
Insights
Quantitative analysis of antigens in tissues is challenging. This study demonstrates that nanoparticle labels with time-resolved fluorescence imaging (TRFI) significantly enhance sensitivity for quantitative immunohistochemistry (IHC).
Area of Science:
- Biomedical imaging
- Biochemistry
- Nanotechnology
Background:
- Tissue and cell examinations offer valuable antigen quantity data.
- Current methods limit truly quantitative analysis in biological samples.
- Previous work established quantitative immunohistochemistry (IHC) with time-resolved fluorescence imaging (TRFI) for linear signal response.
Purpose of the Study:
- To evaluate nanoparticle labels for quantitative IHC.
- To demonstrate the sensitivity and quantitation capabilities of TRFI with nanoparticles.
- To highlight the potential for improved sensitivity in IHC methods.
Main Methods:
- Utilized Eu-chelate containing nanoparticles in the nanometer range as labels for IHC.
- Employed time-resolved fluorescence imaging (TRFI) for detection and quantitation.
- Correlated signal intensity with the amount of bound label.
Main Results:
- Nanoparticles are suitable labels for quantitative IHC.
- TRFI can detect and quantitate single nanoparticle molecules.
- Signal intensity shows strong correlation with bound label amount, improving sensitivity.
Conclusions:
- Nanoparticle-based IHC with TRFI offers enhanced sensitivity for quantitative analysis.
- TRFI is a powerful tool for quantifying antigens or transcripts in tissues and cells.
- This method aids in standardizing and optimizing tissue preparation protocols.
Abstract:
Tissue and cell examinations have a potential to produce extremely valuable information about antigen quantities in samples. Using currently available methods, a truly quantitative analysis is nearly impossible. We have previously shown that immunohistochemical (IHC) detection of prostate-specific antigen and human glandular kallikrein from prostatic tissue, together with time-resolved fluorescence imaging (TRFI), is a suitable method for obtaining quantitative data from biological samples and that the signal response is linear. In this paper we show that Eu-chelate containing particles in the nanometer range are suitable labels for quantitative IHC. Even single nanoparticle molecules can be detected by TRFI and the signals measured can be readily quantitated. The signal intensity correlates very well with the amount of bound label, and the use of nanoparticles could markedly improve the sensitivity of quantitative IHC methods. TRFI provides a powerful tool for providing quantitative data about antigens or transcripts in tissue sections or cultured cells. It is also of major importance in standardization and optimization of protocols for fixation and tissue preparation, including antigen retrieval methods.
