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Updated: Nov 19, 2025

Author Spotlight: Analysis of Fluorescent-Stained Lipid Droplets with 3D Reconstruction for Hepatic Steatosis Assessment

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Improved Folch Method for Liver-Fat Quantification.

Ramgopal Mopuri¹, Mugagga Kalyesubula^1,2, Alexander Rosov¹

¹Volcani Center-Agricultural Research Organization, Institute of Animal Science, Rishon LeZion, Israel.

Frontiers in Veterinary Science

|January 29, 2021

Summary

This summary is machine-generated.

Standardized tissue sonication significantly improves the accuracy and precision of liver-fat quantification in sheep. This enhanced method for determining hepatic steatosis is crucial for diagnosing metabolic diseases in animals and humans.

Keywords:

fatty liver hepatic fat quantification lipids extraction pregnancy toxemia steatosis triglycerides

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Novel In Vivo Micro-Computed Tomography Imaging Techniques for Assessing the Progression of Non-Alcoholic Fatty Liver Disease

Novel In Vivo Micro-Computed Tomography Imaging Techniques for Assessing the Progression of Non-Alcoholic Fatty Liver Disease

Published on: March 24, 2023

Area of Science:

Veterinary Medicine
Biochemistry
Animal Science

Background:

Fatty liver (hepatic steatosis) is a prevalent metabolic disorder in animals and humans.
Accurate quantification of intrahepatic fat is vital for diagnosing and studying liver and metabolic diseases.
Current methods for hepatic steatosis analysis suffer from low reproducibility due to subjective and technical limitations.

Purpose of the Study:

To investigate the effect of standardized tissue sonication on the accuracy and precision of liver-fat determination using the Folch method.
To enhance the efficiency of tissue-lipid extraction for more reliable hepatic fat quantification.
To establish reliable reference values for hepatic fat content in sheep, particularly those with pregnancy toxemia.

Main Methods:

The study utilized liver samples from lean and fatty adult sheep, and pregnant ewes with pregnancy toxemia.
Hepatic fat content was determined using the Folch method, comparing procedures with and without standardized tissue sonication.
Triglyceride levels were quantified enzymatically and correlated with gravimetric results.

Main Results:

Tissue sonication significantly increased the measured hepatic fat content in both lean/fatty lambs (6.6% vs. 5.1%) and ewes with pregnancy toxemia (12.5% vs. 10.8%).
Sonication markedly improved the reproducibility of liver-fat determination, with lower coefficients of variation (CV 3.1% vs. 6.1%).
Enzymatic triglyceride quantification showed moderate correlation with gravimetric analysis (r = 0.632).

Conclusions:

Standardized tissue sonication is a significant improvement for accurate and precise hepatic fat quantification.
The revised protocol enhances lipid extraction efficiency, addressing key limitations in current hepatic steatosis analysis.
The findings provide reliable data for pregnancy toxemic sheep and suggest broad applicability across mammalian species, including humans.