研究目的
To identify a unique biochemical signature for antibody-mediated rejection (AMR) in cardiac allografts using infrared (IR) spectroscopy.
研究成果
IR spectroscopy identified a unique biochemical signature for immunopathologic AMR in cardiac allografts, distinguishing it from nonrejecting allografts and native hearts. This method may facilitate earlier detection of AMR and improve diagnostic accuracy.
研究不足
The study focused on myocardial regions of interest and did not include endothelial regions due to resolution limitations. Future studies will aim to include endothelial regions and examine mixed rejection cases.
1:Experimental Design and Method Selection:
Mid-IR spectroscopy was used to analyze formalin-fixed, paraffin-embedded right ventricular tissue biopsies from posttransplant and native hearts. Principal component analysis coupled with linear discriminant analysis and predictive classifiers were applied to the data.
2:Sample Selection and Data Sources:
A total of 30 posttransplant biopsies (14 positive for C4d and 16 negative for C4d) and 14 native heart biopsies were selected.
3:List of Experimental Equipment and Materials:
Quantum cascade laser (QCL) imaging system (Spero, Daylight Solutions) was used for IR imaging.
4:Experimental Procedures and Operational Workflow:
Tissue sections were deparaffinized, and IR images were acquired. Regions of interest from cardiomyocytes were identified, and spectra were averaged across pixels.
5:Data Analysis Methods:
Principal component analysis coupled with linear discriminant analysis and classification models were used to analyze the spectral data.
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Quantum cascade laser imaging system
Spero
Daylight Solutions
Used for IR imaging of tissue sections.
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Anti-human C4d rabbit polyclonal antibody
Catalog No. 12-5000
American Research Products, Inc.
Used for immunohistochemical staining of C4d.
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Autostainer
BenchMark-XT
Ventana Medical Systems, Inc.
Used for immunostaining.
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HRP diaminobenzidine detection kit
Cat No. 760-500, Ultra-View HRP DAB Kit
Ventana Medical Systems, Inc.
Used for visualization of immunostaining.
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Tissue-Tek Glass Mounting Medium
Sakura Tissue-Tek USA, Inc.
Used for mounting stained sections.
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Aperio ScanScope CS
Leica Biosystems
Used for scanning H&E-stained slides.
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MirrIR low-emissivity reflective slides
Kevley Technologies
Used for mounting tissue sections for IR imaging.
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