{"database":"Pride","file_versions":[{"headers":{"Content-Type":["application/json"]},"body":{"files":{"Xlsx":["ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/07/PXD059505/Identified_proteins_Fig4M.xlsx","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/07/PXD059505/Identified_proteins_Fig4L.xlsx","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/07/PXD059505/Identified_proteins_Fig4F.xlsx","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/07/PXD059505/Identified_proteins_FigS7B.xlsx"],"Other":["ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/07/PXD059505/Mouse_brain_tryptic_digest_10um_Fig4L.imzML","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/07/PXD059505/Mouse_brain_tryptic_digest_10um_Fig4M.ibd","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/07/PXD059505/Mouse_brain_tryptic_digest_10um_FigS7B.imzML","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/07/PXD059505/Mouse_brain_tryptic_digest_10um_Fig4F.ibd","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/07/PXD059505/submission3.px","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/07/PXD059505/Mouse_brain_tryptic_digest_10um_Fig4M.imzML","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/07/PXD059505/Mouse_brain_tryptic_digest_10um_Fig4F.imzML","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/07/PXD059505/Mouse_brain_tryptic_digest_10um_FigS7B.ibd","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/07/PXD059505/Mouse_brain_tryptic_digest_10um_Fig4L.ibd"]},"type":"primary"},"statusCode":"OK","statusCodeValue":200}],"scores":null,"additional":{"labhead_mail":["andreas.roempp@uni-bayreuth.de"],"submitter":["Bastian Jahreis"],"technology_type":["mass spectrometry imaging","Mass Spectrometry","Bottom-up proteomics"],"software":[""],"submitter_keywords":["Spatial proteomics","Tryptic digestion","Ms imaging"],"full_dataset_link":["https://www.ebi.ac.uk/pride/archive/projects/PXD059505"],"tissue":["Brain"],"sample_protocol":["12 µm mouse brain sections were used. Lipids and salts were removed from tissue by several washing steps using ethanol and water. On-tissue digestion was performed by spraying trypsin sequentially on tissue (10 cycles)using a custom-built sprayer. Between each spray, tissue sections were placed inside a custom-built digestion chamber at 40 °C for 10 minutes and incubated for 2 h after the last spray. DHB matrix (30 mg/mL in acetone/water 1:1 + 0.1% TFA (v/v)) was applied using a pneumatic sprayer. MALDI imaging was performed using an AP−SMALDI5 AF (TransMIT GmbH, Giessen, Germany) ion source attached to a Q Exactive HF (Thermo Scientific, Bremen, Germany)."],"repository":["Pride"],"quantification_method":["Not available"],"modification":[""],"data_protocol":["For imaging experiments, Thermo “RAW” data were exported and converted into imzML format using the ‘jimzMLConverter’.ImzML files were processed using MSiReader V.1.02. MALDI images were generated based on m/z values from identified peptides from the protein identification workflow. In addition to the protonated adducts, sodium, potassium and ammonium adducts were calculated and used for image generation. Bin width (m/z window) of the images generated from the imzML dataset was set to ± 3 ppm. Note that all ion images represent raw images which were not normalized, interpolated, or smoothed. Root mean square error (RMSE) as well as the mean ion intensities (without empty pixels) were calculated using in-house software. For LC-MS/MS, peptide extracts were analyzed using an Ultimate 3000 nano-LC system (Thermo Scientific, Bremen, Germany) equipped with an Acclaim PepMap 100 trap column (nanoViper C18, 2 cm length, 100 μm ID) and an EasySpray separation column (PepMap RSLC C18, 50 cm length, 75 μm ID). For peptide separation a flow rate of 250 nL/min and 0.1% formic acid as solvent A was used. The method was based on multi-step gradients from 3% to 25% solvent B (0.1% formic acid in acetonitrile) in 30 min and from 25% to 40% B in 5 min. Data dependent mass spectrometry was performed on a Q Exactive HF-X mass spectrometer (Thermo Scientific, Bremen, Germany) using cycles of one full scan (m/z = 350 to 1600) at FWHM = 60000 at m/z 200 resolution and up to 12 data-dependent MS/MS scans at FWHM = 60000 at m/z 200 resolution. For protein and peptide identification, full scan spectra and MS/MS spectra were searched against the Swiss Prot database using Mascot Search. The “Mus musculus” database was selected and up to one missed cleavage (MC) was allowed. As a fixed modification “Carbamidomethyl (C)” was selected. Methionine oxidation was selected as variable modification. Mass values were set to monoisotopic, and a peptide mass tolerance of ± 10 ppm combined with a fragment mass tolerance of ± 0.02 Da was allowed. Protein identifications were assigned based on a minimum of 1 confident peptide at a false discovery rate of 1% and an ion score of > 50. LC-ESI-MS/MS results from both tissue sections (10 x 1.5 µL: 486 proteins, 10 x 3.0 µL: 538 proteins) were filtered against co-identified proteins, resulting in a final candiate protein list of 365 proteins. Protein identification from MALDI imaging datasets was performed using an in-house developed protein identification toolbox called MSIProteinExplorer. As input, imzML files and FASTA files of the candidate proteins were used. Spatial distributions in the MALDI imaging datasets were identified by unsupervised hierarchical clustering. After clustering, “reasonable spatial distributions” were manually separated from “background” or “noise” resulting in four different structural groups (arbor vitae, granula layer, blood system, noise). These structures were used later for matching against potential peptide distributions. Next, candidate proteins from the LC/MS-MS workflow were digested in-silico and monoisotopic peptide masses were calculated based on possible adducts (H+, Na+, K+, NH4+) Potential peptide ion images were generated at a bin width of Δ m/z 0.01. Several image statistics were calculated based on centroid masses. After that, potential peptide masses were filtered against a maximum tolerated mass error of ± 3 ppm to reduce the number of false positive assignments. Potential peptide ion images were filtered against a minimum of 1 % percentage coverage and a minimum measure of chaos metric of 0.95 to remove empty or noisy images from the dataset56. Remaining potential peptide images for each protein were matched (pearson correlation) against the predefined structural groups which resulted in structural assignments to each protein of the candidate list. In the last step, proteins were identified based on strict identification criteria. In our workflow, possible peptide assignments are based on accurate mass data at MS1 level and spatial distribution patterns. Therefore, strict identification criteria are necessary to account for protein inference: A minimum of two peptides per protein must be detected from which one is unique to the protein database. Additionally, a protein must be assigned to only one pre-defined structural group and 50 % of all remaining peptides per protein must show a consistent spatial distribution (predominant distribution)."],"omics_type":["Proteomics"],"labhead":["Andreas Römpp"],"instrument_platform":[""],"labhead_affiliation":["Bioanalytical Sciences and Food Analysis, University of Bayreuth, Universitaetsstrasse 30, 95447, Bayreuth, Germany"],"submission_type":["PARTIAL"],"species":["Mus Musculus (mouse)"],"publication":["10.1021/ACS.ANALCHEM.6C01111"],"submitter_mail":["bastian.jahreis@uni-bayreuth.de"],"submitter_affiliation":["University of Bayreuth"],"submitter_country":["Germany"],"additional_accession":[]},"is_claimable":false,"name":"MALDI imaging of tryptic peptides in mouse brain measured at 10 µm pixel size","description":"A coronal mouse brain cerebellum section that was measured at 10 µm pixel size revealed detailed histological structures such as granula layer, molecular layer and the blood vessels. These small vessels are only several micrometers in diameter in mouse tissue. MALDI imaging at 10 µm pixel size showed that these blood vessels can be clearly resolved and separated from the surrounding brain areas by MALDI imaging of tryptic peptides. Using an optimized on-tissue digestion approach, up to 283 protein could be tentatively identified in this dataset.","dates":{"publication":"2026-07-13","submission":"2025-01-07"},"accession":"PXD059505","cross_references":{"TAXONOMY":["NEWT:330879","NEWT:377960","NEWT:1131","NEWT:35795","NEWT:2042546","NEWT:259447","NEWT:295546","NCBITaxon:2719036","NEWT:1334565","NEWT:625351","NEWT:258594","NCBITaxon:1280","NEWT:112503","NEWT:1129","NEWT:309807","NEWT:59753","NEWT:89184","NEWT:309800","NEWT:281395","NEWT:1211601","NEWT:876138","NEWT:44271","NEWT:193516","NEWT:2315688","NEWT:1073882","NEWT:6819","NEWT:112509","NEWT:475174","NEWT:111205","NEWT:428386","NEWT:1117","NEWT:498257","NEWT:10036","NEWT:32264","NEWT:12215","NEWT:1590","NEWT:10034","NEWT:33114","NEWT:1596","NEWT:661410","NEWT:638632","NEWT:224326","NEWT:376619","NCBITaxon:79857","NEWT:1096976","NEWT:3765","NEWT:3767","NEWT:1589","NEWT:135622","NEWT:4615","NEWT:672127","NEWT:35786","NEWT:1580","NEWT:399784","NEWT:96731","NEWT:656064","NEWT:1582","NEWT:3760","NEWT:1584","NEWT:36630","NEWT:1585","NEWT:383379","NEWT:418106","NCBITaxon:12721","NEWT:10029","NEWT:913645","NEWT:2492962","NEWT:641809","NEWT:1392488","NEWT:38815","NEWT:317447","NEWT:4688","NEWT:111225","NEWT:7719","NEWT:5530","NEWT:868565","NEWT:135674","NEWT:79329","NEWT:30069","NEWT:12637","NEWT:59729","NEWT:2164133","NEWT:295105","NEWT:108061","NEWT:60711","NEWT:224308","NEWT:3347","NEWT:49928","NEWT:3349","NEWT:160621","NEWT:212790","NEWT:110368","NEWT:55350","NEWT:1310161","NEWT:77133","NEWT:145481","NEWT:1310165","NEWT:29058","NCBITaxon:79824","NEWT:1912919","NEWT:410658","NEWT:5516","NEWT:44688","NEWT:44689","NEWT:498211","NEWT:345076","NEWT:347256","NEWT:5518","NEWT:398007","NEWT:1527468","NEWT:498217","NEWT:498216","NEWT:11320","NEWT:246196","NEWT:246197","NEWT:145458","NEWT:1310173","NEWT:44685","NEWT:161934","NEWT:156587","NEWT:1148","NEWT:5508","NEWT:3329","NEWT:419481","NEWT:5507","NEWT:260499","NEWT:410661","NEWT:1140","NCBITaxon:2157","NEWT:2066563","NEWT:1143","NEWT:1287689","NEWT:1094343","NEWT:1462472","NEWT:1336795","NEWT:644042","NEWT:150808","NEWT:1182590","NEWT:3712","NEWT:3711","NEWT:3714","NCBITaxon:37162","NEWT:270643","NEWT:2065263","NEWT:177437","NEWT:717966","NEWT:10891","NEWT:10418","NEWT:118698","NEWT:1616117","NEWT:118696","NEWT:34865","NEWT:52283","NEWT:28174","NEWT:284812","NEWT:8175","NEWT:43330","NEWT:1603293","NEWT:44664","NEWT:43346","NEWT:3702","NEWT:1245466","NEWT:3704","NEWT:244366","NEWT:1246791","NEWT:176597","NEWT:118694","NEWT:2850","NEWT:118691","NEWT:34871","NEWT:33548","NEWT:52299","NEWT:8165","NEWT:96794","NEWT:3708","NEWT:332648","NEWT:44670","NEWT:926571","NEWT:536231","NEWT:376219","NEWT:219813","NEWT:1510","NEWT:460519","NEWT:1515","NEWT:572307","NEWT:1432138","NEWT:1424507","NEWT:1194599","NEWT:272844","NEWT:1348799","NEWT:33995","NEWT:55784","NEWT:1303443","NEWT:9483","NEWT:485","NEWT:56636","NEWT:2709072","NEWT:2499525","NEWT:2853422","NEWT:487","NEWT:1538553","NEWT:1679718","NEWT:480","NEWT:67767","NEWT:45972","NEWT:46835","NEWT:109757","NEWT:582580","NEWT:1185650","NEWT:216778","NEWT:294607","NEWT:1502","NEWT:128017","NEWT:2836","NEWT:376686","NEWT:237610","NEWT:95486","EFO:0001352","NEWT:119562","NEWT:1883446","NEWT:1233435","NEWT:118223","NEWT:11757","NEWT:987059","NEWT:109760","NEWT:29031","NEWT:317047","NEWT:1509","NEWT:491","NEWT:460523","NEWT:45954","NEWT:235443","NEWT:1579","NEWT:1757312","NEWT:85694","NEWT:109786","NEWT:108458","NEWT:5936","NEWT:321967","NEWT:320637","NEWT:1078283","NEWT:3750","NEWT:983964","NEWT:11706","NEWT:32644","NEWT:527796","NEWT:499175","NEWT:109779","NEWT:3745","NEWT:1715989","NCBITaxon:4751","NEWT:3747","NEWT:1116234","NEWT:1255228","NEWT:410289","NEWT:191701","NEWT:373153","NEWT:1640278","NEWT:472","NEWT:940825","NEWT:29459","NEWT:529507","NEWT:1071661","NEWT:28128","NEWT:470","NEWT:5911","NEWT:748693","NCBITaxon:50557","NEWT:39251","NEWT:29491","NEWT:101841","NEWT:446","NEWT:153481","NEWT:2014887","NEWT:33952","NEWT:11723","NEWT:445","NEWT:153009","NEWT:261756","NEWT:166313","NEWT:63366","NEWT:63367","NEWT:215402","NEWT:1547","NEWT:1290454","NEWT:3726","NEWT:906914","NEWT:81231006","NEWT:28141","NEWT:9031","NEWT:27292","NEWT:1050722","NEWT:3728","NEWT:108931","NEWT:1293497","NEWT:1055524","NEWT:548","NEWT:150475","NEWT:267872","NEWT:381666","NEWT:172269","NEWT:9534","NEWT:2126348","NEWT:5180","NEWT:256737","NEWT:9541","NEWT:8694","NEWT:33936","NEWT:8692","NEWT:2903","NEWT:185579","NEWT:872227","NEWT:13076","NEWT:2587410","NEWT:207340","NEWT:1006581","NEWT:8673","NEWT:33940","NEWT:2126337","NEWT:550","NEWT:33941","NEWT:554","NEWT:451516","NEWT:552","NEWT:1325291","NEWT:42410","NEWT:36185","NEWT:1054211","NEWT:1225786","NEWT:771870","NEWT:51338","NEWT:9995","NEWT:575412","NEWT:28112","NEWT:49012","NEWT:6493","NEWT:10832","NEWT:6494","NEWT:6491","NEWT:507601","NEWT:520","NEWT:64657","NEWT:186441","NEWT:643680","NEWT:214092","NCBITaxon:6157","NEWT:13095","NEWT:162425","NEWT:104105","NEWT:216257","NEWT:9986","NEWT:8654","NEWT:8658","NEWT:1268063","NEWT:8657","NEWT:9983","NEWT:8655","NEWT:5147","NEWT:595495","NE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