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Biology
Utilizzo delle librerie di visione artificiale per semplificare la quantificazione dei nuclei
Utilizzo delle librerie di visione artificiale per semplificare la quantificazione dei nuclei
JoVE Journal
Biology
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JoVE Journal Biology
Using Computer Vision Libraries to Streamline Nuclei Quantification

Utilizzo delle librerie di visione artificiale per semplificare la quantificazione dei nuclei

Full Text
675 Views
06:25 min
June 6, 2025

DOI: 10.3791/67945-v

Danielle E. Levitt1, Alexandra L. Khartabil1, Rylea E. Hall1, Matthew R. DiLeo1, Connor J. Mills1, Ashley K. Williams1, Casey R. Appell2, Ronald G. Budnar, Jr.1, Hui-Ying Luk2

1Metabolic Health & Muscle Physiology Laboratory, Department of Kinesiology and Sport Management,Texas Tech University, 2Applied Exercise Physiology Laboratory, Department of Kinesiology and Sport Management,Texas Tech University

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Please note that some of the translations on this page are AI generated. Click here for the English version.

Overview

This study presents a method for automating the quantification of nuclei in images, which aids in normalizing metabolic data in skeletal muscle research. The automated program, validated across varying cell densities, addresses challenges inherent to manual counting, such as bias and variability.

Key Study Components

Research Area

  • Cell biology
  • Metabolic health
  • Heat therapy effects on muscle

Background

  • Manual quantification of nuclei is time-consuming and biased.
  • Automated methods can improve accuracy and efficiency.
  • Understanding the effects of heat therapy on muscle adaptations is crucial for metabolic health.

Methods Used

  • Automated nuclei counting using an open-source program
  • Cell line models for skeletal muscle research
  • Image processing and analysis

Main Results

  • The automated program showed excellent reliability with an intra-class correlation coefficient of 0.993.
  • Quantification accuracy was high across varying cell densities.
  • Problems with clustering nuclei and artifacts were identified for further improvement.

Conclusions

  • The study demonstrates an objective approach to nuclei quantification that enhances experimental normalization.
  • This method is relevant for advancing research in metabolic health and muscle adaptations.

Frequently Asked Questions

What is the significance of nuclei quantification in this study?
Nuclei quantification is essential for normalizing metabolic data in skeletal muscle research.
How does the automated program compare to manual counting?
The automated program significantly reduces observer bias and variability, showing high reliability against manual counts.
What challenges does manual nuclei counting present?
Manual counting is prone to biases and inconsistencies, making it less reliable than automated methods.
What improvements can be made to the automated method?
Further enhancements can focus on better handling of clustered nuclei and image artifacts.
Is the program user-friendly for those with limited technical skills?
Yes, the program is designed to be accessible for scientists with varying levels of coding experience.
What was the primary validation for the automated program?
The program was validated with high intra-class correlation coefficients across all tested cell densities.
How does this research relate to metabolic health?
It aims to uncover mechanisms of heat therapy in improving muscle and mitochondrial health, particularly in pre-diabetic individuals.

Questo articolo descrive i metodi passo dopo passo per automatizzare la quantificazione dei nuclei basata su immagini utilizzando un programma eseguibile open source convalidato in una gamma di densità cellulari. Questo programma fornisce un'alternativa che affronta le barriere relative ai costi, all'accessibilità per gli utenti con competenze tecnologiche limitate e alla convalida specifica dell'applicazione che può limitare l'utilità delle tecnologie esistenti.

Abbiamo sviluppato questo metodo per normalizzare i dati metabolici provenienti da modelli cellulari per aiutare a identificare i meccanismi, sottolineare gli adattamenti del muscolo scheletrico indotti dalla terapia del calore e, in definitiva, migliorare la salute metabolica tra le persone con pre-diabete.

Dobbiamo contare i nuclei per la normalizzazione sperimentale. La quantificazione manuale dei nuclei presenta sfide tra cui la distorsione dell'osservatore, il tempo e la variabilità nell'imbattersi in campioni o condizioni diverse.

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