Samples on ITO-coated glass slides were sprayed with 1 mL matrix solution (40 mg/mL 2,5-dihydroxybenzoic acid), 20 mM potassium acetate, 70% methanol) (Bruker Daltonics) using a 0.2 mm nozzle caliber airbrush (Procon Boy FWA Platinum; Mr. Hobby, Tokyo, Japan). Distance between the nozzle tip and the slice surface was kept at 10 cm, and spraying was performed for 15 min for uniform matrix deposition. Positive ions of the spinal cord were detected using a MALDI-TOF/TOF-type instrument (ultraflex II TOF/TOF;Bruker Daltonics) equipped with a 355 nm Nd:YAG laser. The laser was set to the minimum spot size with 20% laser power. Mass spectra ranging from mass-to-charge ratio (m/z) 500 to 1000were collected. Laser scan pitch was set to 50 μm. The calibration of m/z values was performed for each IMS measurement using calibration standard substances: 2,5-dihydroxybenzoic acid, bradykinin (Sigma-Aldrich, St Louis, MO, USA), and angiotensin II (Sigma-Aldrich). Regions of interest (ROIs) in spinal cord were determined by comparison with the HE staining result from consecutive tissue sections. Signals were collected using flexControl software (Bruker Daltonics) and reconstruction of ion images (normalized by total ion current) was performed with flexImaging4.0 software (Bruker Daltonics).
Ultraflex 2 tof tof
Manufactured by Bruker
Sourced in Germany
The Ultraflex II TOF/TOF is a high-performance mass spectrometry instrument designed for advanced proteomics and structural biology applications. It combines a time-of-flight (TOF) analyzer with a tandem TOF/TOF configuration, providing high-resolution mass analysis and effective fragmentation capabilities for the detailed characterization of complex samples.
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Lab products found in correlation
12 protocols using ultraflex 2 tof tof
1
MALDI-TOF/TOF Imaging of Spinal Cord Tissue
2
MALDI-TOF-MS Analysis of Conjugates
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Conjugates were analysed by matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF-MS) using an Ultraflex-II TOF/TOF instrument (Bruker Daltonics, Bremen, Germany) equipped with a 200 Hz solid-state Smart beam™ laser. Sinapinic acid was used as the matrix and the samples were spotted using the dried-droplet technique (sample and saturated matrix solution 1:1). The mass spectrometer was operated in the positive linear mode. MS spectra were acquired over an m/z range of 3000–100,000 Da and data was analysed using FlexAnalysis 2.4. software (Bruker, Billerica, MA, USA) provided with the instrument.
3
MALDI-TOF/TOF Imaging and Identification
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A MALDI TOF/TOF-type instrument (Ultraflex II TOF/TOF; Bruker Daltonics) was used for single MS imaging. The instrument was equipped with a 355-nm Nd:YAG laser. The data were acquired in the positive and negative reflectron modes under an accelerating potential of 20 kV using an external calibration method. Signals between m/z of 400 and 1000 were collected, and raster scans on tissue surfaces were performed automatically using FlexControl and FlexImaging 2.0 software (Bruker Daltonics). Laser irradiation consisted of 200 shots per spot. Image reconstructions were performed using FlexImaging 2.0 software. On-tissue molecular identification by MALDI-MS/MS was performed with a MS-IT-TOF (iMScope; Shimadzu Corporation, Kyoto, Japan) system. The molecular weight range for the ion trap was 1.0 Da around the m/z of the precursor ion. Laser irradiation consisted of 200 shots per spot at a 1000 Hz repetition rate. Mass spectra were obtained from a scanning mass range of 100–900 Da.
4
Mass Spectrometry Proteomics Protocol
5
MALDI-TOF/TOF Tissue Analysis Protocol
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The tissue sections were analyzed using a matrix-assisted laser desorption/ionization-time-of-flight/time-of-flight (MALDI-TOF/TOF) type instrument, Ultraflex II TOF/TOF (Bruker Daltonics), which was equipped with a 355 nm smartbeam laser at 200 Hz repetition. Data were acquired in the positive-ion mode using an external calibration method with ions from DHB, angiotensin II, and bradykinin, which covered a mass-to-charge ratio (m/z) range of 100 to 1200. Calibration proteins were deposited on the surfaces of sample sections. Each raster scan was automatically performed in the regions of cancer and normal tissue. The interval between data points was 50 μm, and 100 laser beam shots in a random walk mode for each data point were irradiated. The mass spectrometry parameters were manually optimized to obtain the highest sensitivity, with m/z values in the range of 500–900. All mass spectra were acquired automatically using FlexImaging 2.1 software (Bruker Daltonics).
6
Mass Spectrometry Analysis of Purified Proteins
7
MALDI-TOF/TOF Analysis of Unbound MCs
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To examine the presence of unbound MCs, the tissue samples were also submitted to Ultraflex II™ TOF/TOF (Bruker, Bremen, Germany). Samples aliquots were dissolved in TFA 0.1% and mixed with a saturated matrix solution of α-cyano-4-hydroxycinnamic acid (1:3, v/v) and directly applied onto a target (AnchorChip™, Bruker Daltonics, Billerica, MA, USA). The mass spectrometry was operating in positive reflector mode for MALDI-TOF, or LIFT mode for MALDI-TOF/TOF. Calibration was performed externally with ions of angiotensin I, angiotensin II, substance P, bombesin, insulin b-chain and adrenocorticotropic hormones (clip 1–17 and clip 18–39). Each spectrum was produced by accumulating data from 200 consecutive laser shots.
8
MALDI-TOF-MS analysis of protein masses
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Protein masses were analysed by matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF-MS) using an Ultraflex-II TOF/TOF instrument (Bruker Daltonics, Bremen, Germany) equipped with a 200 Hz solid-state Smart beam laser. The mass spectrometer was operated in the positive linear mode. MS spectra were acquired over an m/z range of 3000–30,000 and data was analyzed using FlexAnalysis 2.4. software provided with the instrument.
Sinapinic acid was used as the matrix (saturated solution in acetonitrile/0.1% trifluoroacetic acid 1:2) and samples were spotted undiluted using the dried-droplet technique. If necessary, samples were diluted in TA33 (33% acetonitrile/0.1% trifluoroacetic acid in water).
Sinapinic acid was used as the matrix (saturated solution in acetonitrile/0.1% trifluoroacetic acid 1:2) and samples were spotted undiluted using the dried-droplet technique. If necessary, samples were diluted in TA33 (33% acetonitrile/0.1% trifluoroacetic acid in water).
9
MALDI-TOF-TOF Imaging of Phospholipids
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We used a MALDI-TOF-TOF-type instrument (ultraflex II TOF/TOF, Bruker Daltonics) to analyze the tissue sections. An Nd:YAG laser (355-nm wavelength) was used for the desorption and ionization of PLs with a 200-Hz repetition rate. The setting parameters for the laser energy, detector gain, and random walk function were optimized manually for each measurement to obtain the highest sensitivity for the m/z range of 460–1,000. The calibration of m/z values detected in each IMS measurement was performed by using Mono DHB ([M + H]+, m/z 155.03), bradykinin ([M + H]+, m/z 757.40), and angiotensin II ([M + H]+, m/z 1046.54) in the positive ion mode. The measurement regions in the War-T and Non-T tissues were automatically scanned by the laser using flexControl (Bruker Daltonics). The number of laser shots at each measurement spot was 200, and the intervals between each measurement point were 200 μm. All data were acquired and visualized by flexImaging 2.1 software (Bruker Daltonics).
10
MALDI-MS/MS Profiling of Brain Lipids
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The molecular characterization was performed with MS/MS and a MALDI linear quadrupole ion-trap mass-spectrometer (Thermo Fisher Scientific Inc., Waltham, MA, USA). The MS/MS analysis was performed directly on the brain sections. The acquisition was in the mid-mass-range mode (m/z 100–1,000), which is the positive-ion detection mode, with an ionization voltage of 30 V and a collision voltage of 35 V. The PC molecular species were identified from the neutral loss compositions, which were determined from the deltas of the precursor and product ions in the MS/MS spectra.
The IMS of the identified PC species was performed with a MALDI time-of-flight (TOF)/TOF-type instrument (Ultraflex II TOF/TOF; Bruker Daltonics GmbH) that was equipped with a 355-nm Nd:YAG laser. The data were acquired in the positive-ion reflector mode under an accelerating potential of 20 kV. Calibration was performed with an external calibration method. The signals of m/z = 700–1,000 were measured. Raster scans on the tissue surfaces were performed automatically with FlexControl and FlexImaging 2.0 software (Bruker Daltonics GmbH). The number of laser irradiations was 200 shots per spot. Image reconstruction was performed with FlexImaging 2.0 software.
The IMS of the identified PC species was performed with a MALDI time-of-flight (TOF)/TOF-type instrument (Ultraflex II TOF/TOF; Bruker Daltonics GmbH) that was equipped with a 355-nm Nd:YAG laser. The data were acquired in the positive-ion reflector mode under an accelerating potential of 20 kV. Calibration was performed with an external calibration method. The signals of m/z = 700–1,000 were measured. Raster scans on the tissue surfaces were performed automatically with FlexControl and FlexImaging 2.0 software (Bruker Daltonics GmbH). The number of laser irradiations was 200 shots per spot. Image reconstruction was performed with FlexImaging 2.0 software.
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