By Michael E. Himmel
Biomass conversion learn is a mixture of easy technology, utilized technology, and engineering checking out and research. Conversion technological know-how contains the preliminary therapy (called pre-treatment) of the feedstock to render it extra amenable to enzyme motion, enzymatic saccharification, and eventually product formation via microbiological or chemical approaches. In Biomass Conversion: equipment and Protocols, expert researchers within the box aspect equipment that are now usual to check biomass conversion. those equipment comprise Biomass Feedstocks and Cellulose, Plant cellphone Wall Degrading Enzymes and Microorganisms, and Lignins and Hemicelluloses. Written within the hugely winning Methods in Molecular Biology™ sequence layout, the chapters contain the type of certain description and implementation recommendation that's an important for buying educated, reproducible leads to the laboratory.
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Extra info for Biomass Conversion: Methods and Protocols
Gov/ij/). , SanJose, CA). ● Osirix software for 3D visualization and analysis (http://www. com/). edu/chimera/). 3. 1. Fixation and Embedding by Microwave Processing 1. This protocol has been modified from those developed and used by Elaine Humphrey, and Lacey Samuels at the University of British Columbia and others (7–9). See Note 1 for how to choose between chemical and cryo-fixation methods. 2. All steps are carried out in the PELCO laboratory microwave unless stated otherwise (Fig. 2a). 3. 8 mL Eppendorf® or other suitable micro tube.
2. Conductive target material should have good electrical conductivity, high secondary electron emission, considered electron dense, non-oxide forming and maintain a nongranular structure as a thin layer. Good choices of conductive coatings include gold (Au) and Pt/Pd as they inherently possess four of the criteria listed; however, at high resolution FE-SEM imaging some granularity may be detected. 3. Samples should be secured to aluminum stubs with doublesided carbon tape in a manner that maximizes contact with the holder.
2. Critical Point Drying 1. Critical point drying (CPD) is utilized to reduce the surface tension of drying to zero, preventing deformation and collapse of specimen structure. Recommended for samples that are sensitive to ice crystal formation and subsequent damage (Fig. 2c). 2. Wash fixed or non-fixed samples in water (5 min; 2×). 3. Dehydrate using a series of ethanol washes (a) 50% ETOH and 50% water (5 min). (b) 75% ETOH and 25% water (5 min). (c) 95% ETOH and 5% water (5 min). (d) 100% ETOH (5min; 3×).