Ebook: Biotechnology Applications of Microinjection, Microscopic Imaging, and Fluorescence

Individual cells behave in surpnsmg ways that cannot be deduced from the averaged results of an organ as assessed by the use of conventional biochemical methods. Thus multicellular plant and animals systems are being investigated by an increasing array of histochemical and cytochemical techniques based on general chemical or specific immunological interactions to identify structural materials and to assess biological activities. In recent years there has been an increasing range of fluorescent probes, along with advanced computerised imaging and analysis techniques, which allows the behaviour of individual living cells to be followed in considerable detail. The parallel use of microinjection, microelectrodes and patch-clamping provides additional information about cells and their responses. Recombinant DNA technology has highlighted the desirability and the power of microinjecting defined materials into specific cells and so manipulating their fundamental biochemistry. New hypotheses are being tested which will form the cornerstone of future developments across the whole spectrum of biotechnology. The First European Workshop on Biotechnology Applications of Microinjection, Microscopic Imaging and Fluorescence was run at the University of East London, U.K, 21st-24th April, 1992 with the objective of bringing together a diverse group of individuals who were using these state-of-the-art applications for biotechnological exploration. A novel feature of the meeting was paiticipation by instrument manufacturers in the programme: there were hands-on workshops (where living cells could be examined), combined with the poster sessions.

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Microinjection (G. Sczakiel et al.). How to Make Glass Microtools for the Injection of Isolated Plant Sperm Cells into Embryo Sac Cells Using a Microforge (C.J. Keijzer). Application of Confocal Microscopy for the Study of Neuronal Organization in Human Cortical Areas after Microinjection of Lucifer Yellow (P.V. Belichenko et al.). Flourescent Probes (R.W. Horobin, F. RashidDoubell). Flourescent Phospholipids in Membrane and Lipoprotein Research (A. Hermetter et al.). Selection of Flourescent Golgi Complex Probes Using StructureActivity Relationship Models (F. RashidDoubell, R.W. Horobin). WholeCell PatchClamping (B. Van Duijn et al.). Flourescent Analysis of Replication and Intermediates of Chromatin Folding in Nuclei of Mammalian Cells (G. Banfalvi). Autoflourescence in Potato Tuber Phellem (T. Hendriks et al.). Quantitative Localization of HIV Proteins in Mammalian Cells (L.E.A. Amet et al.). Concepts of the Cytometric Approach (S.C. Brown). 14 additional articles. Index.

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Microinjection (G. Sczakiel et al.). How to Make Glass Microtools for the Injection of Isolated Plant Sperm Cells into Embryo Sac Cells Using a Microforge (C.J. Keijzer). Application of Confocal Microscopy for the Study of Neuronal Organization in Human Cortical Areas after Microinjection of Lucifer Yellow (P.V. Belichenko et al.). Flourescent Probes (R.W. Horobin, F. RashidDoubell). Flourescent Phospholipids in Membrane and Lipoprotein Research (A. Hermetter et al.). Selection of Flourescent Golgi Complex Probes Using StructureActivity Relationship Models (F. RashidDoubell, R.W. Horobin). WholeCell PatchClamping (B. Van Duijn et al.). Flourescent Analysis of Replication and Intermediates of Chromatin Folding in Nuclei of Mammalian Cells (G. Banfalvi). Autoflourescence in Potato Tuber Phellem (T. Hendriks et al.). Quantitative Localization of HIV Proteins in Mammalian Cells (L.E.A. Amet et al.). Concepts of the Cytometric Approach (S.C. Brown). 14 additional articles. Index.
Content:
Front Matter....Pages i-xi
Microinjection: A Technique to Study Inhibition of HIV-1 Replication Mediated by Antisense RNA and Parvovirus Genes....Pages 1-10
How to Make Glass Microtools for the Injection of Isolated Plant Sperm Cells into Embryo SAC Cells Using a Microforge....Pages 11-17
Application of the Firefly Luciferase Reporter Gene to Microinjection Experiments in Xenopus Oocytes....Pages 19-28
Application of Confocal Microscopy for the Study of Neuronal Organization in Human Cortical Areas after Microinjection of Lucifer Yellow....Pages 29-35
The Dye-Transfer Assay Permits the Evaluation of the Modulation of Junctional Communication by Chemicals: Results Obtained Employing a Highly Sensitive Video-Recording System Connected with the Microinjector....Pages 37-43
Comparison of Fluorescent Dye Transfer in Intercellular Communication Studies Using Scrape-Loading and Microinjection Techniques....Pages 45-52
Characterization of the Zea Mays Embryo Sac Using Fluorescent Probes and Microinjection of Lucifer Yellow into the Female Cells....Pages 53-60
Fluorescent Probes — Where Do they Go in Cells, and Why?....Pages 61-66
Fluorescent Phospholipids in Membrane and Lipoprotein Research....Pages 67-72
Selection of Fluorescent Golgi Complex Probes Using Structure-Activity Relationship Models....Pages 73-78
Whole-Cell Patch-Clamping: Introducing Substances into Cells During Electrical Measurements from the Cell Membrane -A Review of Potential Difficulties in Plant and Animal Cells....Pages 79-97
Fluorescent Analysis of Replication and Intermediates of Chromatin Folding in Nuclei of Mammalian Cells....Pages 99-109
Autofluorescence in Potato Tuber Phellem: Location and Time Course of Appearance During Tuber Formation....Pages 111-119
Flow Cytometric Assessment of Gentamicin Nephrotoxicity in Established Renal Cell Lines....Pages 121-127
The Use of the Fluorescent Probe Fura-2 For Intracellular Free Calcium Measurements: Some Methodological Aspects....Pages 129-132
Studies on the Effects of Peroxisome Proliferators as Novel Ca2+ Mobilizing Agents Using Indo-1-Loaded Hepatocytes....Pages 133-142
Confocal Fluorescence Ratio Imaging of pH in Plant Cells....Pages 143-151
Analyses of the Calcium and Phosphorus Distribution in Predentine Tissue of Rat Incisors....Pages 153-163
Quantitative Localisation of HIV Proteins in Mammalian Cells....Pages 165-171
Concepts of the Cytometric Approach....Pages 173-184
Laser Microperforation of Medicago Sativa Root Hair Cells....Pages 185-196
Application of pH-Sensitive Fluorescent Dyes to Fungi....Pages 197-205
Very Low Level Fluorescence Detection and Imaging Using a Long Exposure Charge Coupled Device System....Pages 207-219
Modulation of Nucleus Formation in Mouse Oocytes Fused with Early and Late G2 Blastomeres....Pages 221-230
Back Matter....Pages 231-237
....Pages 239-255