Cell sampling and analysis (SiCSA): metabolites measured at single cell resolution.
By using a fine oil-filled glass microcapillary mounted on a micromanipulator, the solutes of individual plant cells can be sampled. These samples can then be analysed using a range of physical and chemical methods. Hydrostatic pressure (cell pressure probe), osmotic pressure (picolitre osmometer), organic solutes (enzyme-linked fluorescence microscope spectrometry or capillary electrophoresis), inorganic solutes (X-ray microdroplet analysis or capillary electrophoresis), (14)C (mass spectrometry), proteins (microdroplet immunoblotting), and mRNA (rt PCR) have been measured. Collectively, the battery of techniques is called single cell sampling and analysis (SiCSA) and all of the techniques have relevance to the study of plant metabolism at the resolution of the individual cell. This review summarizes the techniques for SiCSA and presents examples of applications used in this laboratory, in particular those relating to cell metabolism.
Tomos
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Ysgol Gwyddorau Biolegol, Prifysgol Cymru Bangor, Bangor, Gwynedd LL57 2UW, Wales, UK. a.d.tomos@bangor.ac.uk
Sharrock
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D002477:Cells / Q000737:chemistry; D002623:Chemistry Techniques, Analytical / Q000295:instrumentation / Q000379:methods*; D004577:Electron Probe Microanalysis; D019075:Electrophoresis, Capillary; D007287:Inorganic Chemicals / Q000032:analysis; D008846:Micromanipulation / Q000295:instrumentation / Q000379:methods*; D009930:Organic Chemicals / Q000032:analysis; D009997:Osmotic Pressure; D059828:Plant Cells; D010940:Plant Proteins / Q000032:analysis; D010944:Plants / Q000737:chemistry / Q000378:metabolism; D012333:RNA, Messenger / Q000032:analysis; D018749:RNA, Plant / Q000032:analysis; D013050:Spectrometry, Fluorescence; D013395:Sucrose / Q000378:metabolism
D007287:Inorganic Chemicals; D009930:Organic Chemicals; D010940:Plant Proteins; D012333:RNA, Messenger; D018749:RNA, Plant; D013395:Sucrose