MALDI-TOF MS Profiling of Microbial Samples

Microbial colony smears or liquid samples were applied to a target location on a stainless steel MALDI plate. The target plate was incubated in a 70% citric acid buffer in a humidified chamber for 30 minutes at 110ºC. The MALDI plate was washed with deionized water from a squeeze bottle, allowed to air dry, then 1 μL of Norharmane matrix solution was applied (10 mg/mL in 12:6:1, v/v/v chloroform/methanol/water) to each target location (36 (link)). Following the method of Leung et al. (52 (link)), spectra were acquired from target locations in negative ion mode using a Microflex LRF MALDI-TOF MS (Bruker, Billerica MA) in reflectron mode with a limited mass range of 1,000 – 2,400 m/z. Typically, 300 laser shots were summed to acquire each mass spectrum.

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Gauthier A.E., Chandler C.E., Poli V., Gardner F.M., Tekiau A., Smith R., Bonham K.S., Cordes E.E., Shank T.M., Zanoni I., Goodlett D.R., Biller S.J., Ernst R.K., Rotjan R.D, & Kagan J.C. (2021). Deep-sea microbes as tools to refine the rules of innate immune pattern recognition. Science immunology, 6(57), eabe0531.

Publication 2021

Microflex LRF MALDI-TOF MS

Buffer Chloroform Citric acid Maldi ms Mass spectrum Methanol Norharmane Stainless steel

Corresponding Organization : Boston University

Other organizations : University of Maryland, Baltimore, Ministry of Fisheries and Marine Resource Development, Wellesley College, Temple University, Woods Hole Oceanographic Institution, University of Gdańsk

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Variable analysis

independent variables

Incubation of the MALDI plate in a 70% citric acid buffer in a humidified chamber for 30 minutes at 110ºC

dependent variables

Microbial colony smears or liquid samples applied to the MALDI plate
Mass spectra acquired from the target locations in negative ion mode using a Microflex LRF MALDI-TOF MS

control variables

Stainless steel MALDI plate
Norharmane matrix solution (10 mg/mL in 12:6:1, v/v/v chloroform/methanol/water) applied to each target location
Limited mass range of 1,000 – 2,400 m/z for mass spectra acquisition
300 laser shots summed to acquire each mass spectrum

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