The analysis of gene expression in developing organs is a valuable tool for the assessment of genetic fingerprints during the various stages of differentiation. Complex processes in developing tissues are particularly difficult to understand in terms of biochemical phenomena. Laser-assisted microdissection (LAM) allows the efficient and precise capture of cells or groups of cells from developing tissues in sufficient quantities and within the context of time and space to permit the subsequent molecular characterization of the targeted tissue. The technique development has dramatically increased the ease of isolating specific cells which, together with progress in tissue preparation and microextraction protocols, allows for broad-range down-stream applications in the fields of genomics, transcriptomics and proteomics. This review gives an overview of the LAM technology and its application in developmental biology.
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http://dx.doi.org/10.1002/jez.b.21133 | DOI Listing |
J Biol Chem
December 2024
Laboratory of Reproductive Neurobiology, HUN-REN Institute of Experimental Medicine, Budapest, 1083 Hungary. Electronic address:
We developed a versatile 'IHC/LCM-Seq' method for spatial transcriptomics of immunohistochemically detected neurons collected with laser-capture microdissection (LCM). IHC/LCM-Seq uses aluminon and polyvinyl sulfonic acid for inventive RNA-preserving strategies to maintain RNA integrity in free-floating sections of 4% formaldehyde-fixed brains. To validate IHC/LCM-Seq, we first immunostained and harvested striatal cholinergic interneurons with LCM.
View Article and Find Full Text PDFThe ability to bring spatial resolution to omics studies enables a deeper understanding of cell populations and interactions in biological tissues. In the case of proteomics, single-cell and spatial approaches have been particularly challenging, due to limitations in sensitivity and throughput relative to other omics fields. Recent developments at the level of sample handling, chromatography, and mass spectrometry have set the stage for proteomics to be established in these new disciplines.
View Article and Find Full Text PDFPLoS 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.
View Article and Find Full Text PDFExpert Rev Proteomics
December 2024
Biological Sciences Division, Pacific Northwest National Laboratory, Richland, Washington, USA.
Introduction: Spatial biology is an emerging interdisciplinary field facilitating biological discoveries through the use of spatial omics technologies. Recent advancements in spatial transcriptomics, spatial genomics (e.g.
View Article and Find Full Text PDFZhongguo Zhong Yao Za Zhi
October 2024
School of Traditional Chinese Medicine, Guangdong Pharmaceutical University Guangzhou 510006, China State Key Laboratory for Quality Ensurance and Sustainable Use of Dao-di Herbs, National Resource Center for Chinese Materia Medica, China Academy of Chinese Medical Sciences Beijing 100700, China Key Laboratory of Biology and Cultivation of Herb Medicine, Ministry of Agriculture and Rural Affairs Beijing 100700, China.
Laser capture microdissection(LCM) combined with gas chromatography-mass spectrometry(GC-MS) was employed to investigate the distribution of volatile compound in the secretory cavities of different tissues(cortex, phloem, xylem, and pith) in different rhizome sections of wild Atractylodes lancea. The same method was used to study the metabolic characteristics of the volatile compounds in the secretory cavities of different rhizome tissues of A. lancea transplanted for 1-3 years.
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