Maldi biotyper compass 4

Fresh colony yeast was applied directly to a sample plate of the equipment, the samples were coated with a saturated solution of α-cyano acid 4-hydroxy cinnamic diluted in 50% ACN with 2.5% TFA. The Mass spectra were recorded using a MALDI-TOF MS Autoflex Speed (Bruker Daltonics, Bremen, Germany) equipped with an intelligent beam laser source (334 nm). The analyses were analyzed in linear mode with positive polarity, acceleration voltage of 20 kV and delay extraction of 220 ns. Each spectrum was collected as an average of 1200 laser shots with enough energy to produce good spectra without saturation in the range of m/z from 2000 to 20,000. Before analysis, the equipment was externally calibrated with the protein calibration standard I (Bruker Daltonics, Bremen, Germany, insulin, ubiquitin, cytochrome C and myoglobin) with FlexControl 1.4 software (Bruker Daltonics, Bremen, Germany). The analyses were analyzed with the MALDI Biotyper Compass 4.1 software (Bruker Daltonics, Bremen, Germany) in the range of m/z 3000–15,000 compared with a library of 1301 spectra of fungal identifications.

Samples of yeast were incubated in the dark at 37 °C for 48 h, then colony samples were applied directly to a sample plate of the equipment, following the procedures as described by Gobom et al., 2011 [34 (link)], where the sample was coated with a saturated solution of α-cyano acid 4-hydroxy cinnamic diluted in 50% ACN with 2.5% TFA. The mass spectra were used using a MALDI-TOF-MS Autoflex Speed (Bruker Daltonics, Bremen, Germany) equipped with an intelligent beam laser source (334 nm). Analysis was carried out in the linear mode with positive polarity, an acceleration voltage of 20 kV and a delay extraction of 220 ns. Each spectrum was collected as an average of 1200 laser shots with enough energy to produce good spectra without saturation in the range of m/z from 2000 to 20,000. Before analysis, the equipment was externally calibrated with the protein calibration standard I (Bruker Daltonics, Bremen, Germany; insulin, ubiquitin, cytochrome C and myoglobin) with FlexControl 1.4 software (Bruker Daltonics, Bremen, Germany). Analysis was carried out with the MALDI Biotyper Compass 4.1 software (Bruker Daltonics, Bremen, Germany) in the range of m/z 3000–15,000 compared to a library of 1301 spectra of fungal identifications.

For MALDI-TOF/TOF MS analysis, 1 µL of the dry C11 and C27 strains was applied on the equipment plate (Bruker Daltonics, Bremen, Germany) and coated with 1 µL of a saturated solution of α-cyano 4-hydroxy cinnamic acid. Mass spectra were obtained with a smart beam laser source (334 nm). The analyses were performed with positive polarity, 20 kV acceleration voltage, and extraction with a 220 ns delay. Each spectrum was collected as an average of 1200 laser shots in the range of 2000 to 20,000 m/z. The equipment was calibrated with a protein calibration standard I (insulin, ubiquitin, cytochrome C, and myoglobin). The strain analyses were performed with the MALDI Biotyper Compass 4.1 software (Bruker Daltonics, Bremen, Germany) in the range of 3000–15,000 m/z. A score of ≥2 denoted identifications to the species level, and an intermediate log score between <2 and ≥1.7 denoted identification to the genus level.