Improved Dispersion of Bacterial Endospores for Quantitative Infrared Sampling on Gold Coated Porous Alumina Membranes

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Abstract:

An improved method for qualitative and quantitative sampling of bacterial endospores using Fourier transform infrared (FT-IR) microscopy on gold-coated porous alumina membranes is presented. Bacillus subtilis endospores were filtered onto gold-coated alumina membranes serving as substrates. Studies in the mid-infrared (MIR) region revealed the characteristic bacterial absorption spectrum at low surface concentration, while scanning electron microscopy (SEM) images of the same samples provided precise calculation of the surface concentration of the bacterial endospores. Under the conditions of study, the average concentration of endospores was determined to be 1356 ± 35 spores in a 100 × 100 μm2 area, with a relative standard deviation of 0.0260. Examination of ten random spots on multiple substrates with FT-IR microscopy apertured to the same area gave an average relative standard deviation of 0.0482 in the signal strength of the amide A band at 3278 cm−1. An extinction cross-section in reflection of σext = (7.8 ± 0.6) × 10−9 cm2/endospore was calculated for the amide A band at the frequency of its peak absorbance, 3278 cm−1. The absorption cross-section of the amide A band in reflection is estimated to be σabs ≈ (2.10 ± 0.12) × 10−9 cm2/endospore.

Keywords: ABSORPTION CROSS-SECTION; ALUMINA MEMBRANE; ANODISCTRADE; ANOPORETRADE; BACTERIAL ENDOSPORES; FOURIER TRANSFORM INFRARED MICROSPECTROSCOPY; FT-IR MICROSPECTROSCOPY; MID-INFRARED; MIR; REFLECTANCE

Document Type: Research Article

DOI: http://dx.doi.org/10.1366/0003702054615188

Affiliations: 1: Science and Technology Corp., 10 Basil Sawyer Drive, Hampton, Virginia 23666; Department of Chemistry and Biochemistry, University of South Carolina, Columbia, South Carolina 29208 2: Department of Chemistry and Biochemistry, University of South Carolina, Columbia, South Carolina 29208 3: Department of Molecular, Microbial and Structural Biology, University of Connecticut Health Center, Farmington, Connecticut 06030 4: U.S. Army Edgewood Chemical Biological Center, Aberdeen Proving Ground, Maryland 21010 5: Department of Chemistry, The Johns Hopkins University, Baltimore, Maryland 21218

Publication date: August 1, 2005

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