Multiphoton Microscopy and Mass Spectrometry for Revealing Metabolic Heterogeneity of Hepatocytes in vivo

Abstract

The aim of the investigation was to study the possibility of revealing the heterogeneity of normal liver hepatocytes in terms of metabolic status using the modern methods of multiphoton microscopy and mass spectrometry.Materials and Methods. Heterogeneity of hepatocytes in terms of total metabolic activity was assessed using multiphoton microscopy based on the autofluorescence intensity of intracellular cofactors NAD(P)H and FAD. Hepatocyte heterogeneity in terms of intensity of intracellular metabolic processes was determined using the fluorescence lifetime imaging (FLIM) method based on the data about fluorescence lifetime contributions of various forms of NAD(P)H. The method of time-of-flight secondary ion mass spectrometry (TоF-SIMS) was used to study the lipid and amino acid composition of hepatocytes.Results. It has been revealed using multiphoton microscopy that hepatocytes are heterogeneous in terms of general metabolic activity. Using FLIM, it was established that the heterogeneity degree was high in terms of intensity of oxidative phosphorylation, glycolysis,

Spatial metabolomics and its application in the liver

“…Topographical distributions of liver metabolic processes have been recently re-examined by LCM approaches coupled with RNA sequencing to provide a comprehensive transcriptome analysis of human and mouse liver zonation 40 . In addition, a recent study using MSI has shown hepatocyte heterogeneity in terms of amino acid and lipid composition 69 . Also, using MSI, liver metabolome was shown to change with fasting in a localized pattern, including increased levels of fatty acids and tricarboxylic acid cycle intermediates 70 …”

“… 40 In addition, a recent study using MSI has shown hepatocyte heterogeneity in terms of amino acid and lipid composition. 69 Also, using MSI, liver metabolome was shown to change with fasting in a localized pattern, including increased levels of fatty acids and tricarboxylic acid cycle intermediates. 70 …”

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