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.

Table of Contents:
Microinjection: A Technique to Study Inhibition of HIV-1 Replication Mediated by Antisense RNA and Parvovirus genes.- How to Make Glass Microtools for the Injection of Isolated Plant Sperm Cells into Embryo Sac Cells Using a Microforge.- Application of the Firefly Luciferase Reporter Gene to Microinjection Experiments in Xenopus Oocytes.- Application of Confocal Microscopy for the Study of Neuronal Organization in Human Cortical Areas After Microinjection of Lucifer Yellow.- 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.- Comparison of Fluorescent Dye Transfer in Intercellular Communication Studies Using Scrape-Loading and Microinjection Techniques.- Characterization of the Zea mays Embryo Sac Using Fluorescent Probes and Microinjection of Lucifer Yellow into the Female Cells.- Fluorescent Probes — Where Do They Go in Cells, and Why?.- Fluorescent Phospholipids in Membrane and Lipoprotein Research.- Selection of Fluorescent Golgi Complex Probes Using Structure-Activity Relationship Models.- Fluorescent Probes for the Assessment of the Subcellular Effects of Chemicals and Disease Processes In Vivo and In Vitro.- 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.- Fluorescent Analysis of Replication and Intermediates of Chromatin Folding in Nuclei of Mammalian Cells.- Autofluorescence in Potato Tuber Phellum: Location and Time Course of Appearance During Tuber Formation.- Flow Cytometric Assessment of Gentamicin Nephrotoxicity in Established Renal Cell Lines.- The Use of the FluorescentProbe Fura-2 for Intracellular Free Calcium Measurements: Some Methodological Aspects.- Studies on the Effects of Peroxisome Proliferators as Novel Ca2+ Mobilizing Agents using Indo-1-Loaded Hepatocytes.- Confocal Fluorescence Ratio Imaging of pH in Plant Cells.- Analysis of the Calcium and Phosphorus Distribution in Predentine Tissue of Rat Incisors.- Quantitative Localisation of HIV Proteins in Mammalian Cells.- Concepts of the Cytometric Approach.- Laser Microperforation of Medicago sativa Root Hair Cells.- Application of pH-sensitive Fluorescent Dyes to Fungi.- Very Low Level Fluorescence Detection and Imaging Using a Long Exposure Charge Coupled Device System.- Modulation of Nucleus Formation in Mouse Oocytes Fused with Early and Late G2 Blastomeres.