Physical Analysis Group
Feedstocks engineering, optimization of deconstruction processes (i.e., chemical, enzymatic, or microbial pretreatment and saccharification approaches), and fuels pathway re-engineering benefit from a variety of biophysical techniques that allow monitoring and quantification of the results of various experimental designs.
These biophysical techniques include: thermo gravimetric analysis (TGA), differential scanning calorimetry (DSC), pyro gas chromatography mass spectrometry (GC/MS), mechanical stress analysis, and a variety of vibrational/optical spectroscopic and imaging techniques (i.e., Fourier Transform Infra Red (FTIR), Raman Scattering, Polarized Raman Scattering, Circular Dichroism, and fluorescence analysis). Such techniques allow us to obtain information about compositional, phase, and molecular weight changes of biopolymers, as well as their degree of organization in plant cell walls.
Figure 1. TEM imaging for precision measurements of plant cell walls (close-up).
Our unique expertise is in mechanical strength testing of mutants as part of a high-throughput pipeline, faithful sample preservation, data collection, 3D reconstruction and quantitative analysis using high-resolution imaging electron microscopy (including cryogenic and microwave-assisted processing), and 2D and 3D transmission and scanning electron microscopy (electron tomography and focused ion beam scanning electron microscopy [FIB/SEM]).
Figures 2a-d. SEM imaging of individual bacteria and lignocellulose-degrading microbial communities.
Our target samples include: in-depth characterization of feedstocks cell wall mutants; monitoring of cell fractionation protocols; monitoring of the efficacy of the pretreatment as well as chemical, enzymatic, and microbial deconstruction process on various biomass; and, assessment of protein abundance and subcellular localization on microbial host systems. Moreover, we have begun to study in fair detail the lignocellulosic biomass deconstruction process in model microbial communities.
Figure 3. Monitoring pretreatment and enzymatic deconstruction of biomass using SEM imaging.
We are engaged in a large number of collaborative projects within JBEI. We provide quantification of a variety of tantalizing observations, advancing the scientific community's understanding of the physical, chemical, and biological processes that are central to lignocellulosic biofuel production.
