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Deciphering cellular complexity: advances and future directions in single-cell protein analysis.
Front Bioeng Biotechnol
January 2025
Yunnan Key Laboratory of Cell Metabolism and Diseases, Yunnan University, Kunming, China.
Single-cell protein analysis has emerged as a powerful tool for understanding cellular heterogeneity and deciphering the complex mechanisms governing cellular function and fate. This review provides a comprehensive examination of the latest methodologies, including sophisticated cell isolation techniques (Fluorescence-Activated Cell Sorting (FACS), Magnetic-Activated Cell Sorting (MACS), Laser Capture Microdissection (LCM), manual cell picking, and microfluidics) and advanced approaches for protein profiling and protein-protein interaction analysis. The unique strengths, limitations, and opportunities of each method are discussed, along with their contributions to unraveling gene regulatory networks, cellular states, and disease mechanisms.
View Article and Find Full Text PDFInvestigation of Exome-Wide Tumor Heterogeneity on Colorectal Tissue-Based Single Cells.
Int J Mol Sci
January 2025
Department of Internal Medicine and Oncology, Faculty of Medicine, Semmelweis University, 1085 Budapest, Hungary.
The progression of colorectal cancer is strongly influenced by environmental and genetic conditions. One of the key factors is tumor heterogeneity which is extensively studied by cfDNA and bulk sequencing methods; however, we lack knowledge regarding its effects at the single-cell level. Motivated by this, we aimed to employ an end-to-end single-cell sequencing workflow from tissue-derived sample isolation to exome sequencing.
View Article and Find Full Text PDFEffects of periodontal disease on the proteomic profile of the periodontal ligament.
J Proteomics
January 2025
Department of Diagnosis and Surgery, School of Dentistry at Araraquara, São Paulo State University - UNESP, Araraquara, São Paulo, Brazil; Department of Periodontology and Operative Dentistry, University Medical Center of the Johannes Gutenberg University, Mainz, Germany.
Periodontal disease affects over 1 billion people globally. This study investigated how periodontitis affects the protein profile of the periodontal ligament (PDL) in rats. Eight Holtzman rats were divided into control and experimental periodontitis groups.
View Article and Find Full Text PDFAcquisition and transcriptomic analysis of tissue micro-regions using a capillary-based method.
J Pharm Biomed Anal
March 2025
State Key Laboratory of Digital Medical Engineering, School of Biological Science & Medical Engineering, Southeast University, Nanjing, Jiangsu, China. Electronic address:
Profiling the site-specific transcriptomes of microregions of interest (mROIs) contributes to a complete understanding of multicellular organisms. However, the simple and efficient isolation of mROIs for spatially detecting gene expression remains challenging. Here, we develop an efficient capillary-based microdissection system (CMS) for precisely isolating targeted samples from tissue sections.
View Article and Find Full Text PDFDevelopment of a sequential laser microdissection tissue cuts workflow for the spatial and quantitative analysis of drugs in fresh frozen tissue sections.
PLoS One
December 2024
Faculty of Health, Maastricht MultiModal Molecular Imaging Institute (M4I), Medicine and Life Sciences, Maastricht University, Maastricht, The Netherlands.
Mass spectrometry imaging (MSI) is a well-established technique that allows to determine the distribution of small molecules, such as lipids, metabolites, and drugs, as well as large molecules in tissue sections. Because of the tissue heterogeneity, resulting in different matrix effects, and to the fact that the measured compounds are not entirely "extracted" from the tissue during the measurement, the absolute quantitative aspect of MSI is limited. To combine compound quantification with spatial information on fresh frozen unstained tissue sections, laser (capture) microdissection has been used to isolate tissue sections for compound extraction and LC-MS/MS quantification.
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