{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,3]],"date-time":"2026-06-03T02:13:45Z","timestamp":1780452825524,"version":"3.54.1"},"reference-count":34,"publisher":"MDPI AG","issue":"21","license":[{"start":{"date-parts":[[2022,10,26]],"date-time":"2022-10-26T00:00:00Z","timestamp":1666742400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Advanced Research and Development Programs for Medical Innovation (AMED-CREST)","award":["AMED JP17gm0910001"],"award-info":[{"award-number":["AMED JP17gm0910001"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Cells"],"abstract":"<jats:p>Space and time coherent mapping (STCM) is a technology developed in our laboratory for improved matrix-assisted laser desorption ionization (MALDI) time of flight (TOF) imaging mass spectrometry (IMS). STCM excels in high spatial resolutions, which probe-based scanning methods cannot attain in conventional MALDI IMS. By replacing a scanning probe with a large field laser beam, focusing ion optics, and position-sensitive detectors, STCM tracks the entire flight trajectories of individual ions throughout the ionization process and visualizes the ionization site on the sample surface with a subcellular scale of precision and a substantially short acquisition time. Results obtained in thinly sectioned leech segmental ganglia and epididymis demonstrate that STCM IMS is highly suited for (1) imaging bioactive lipid messengers such as endocannabinoids and the mediators of neuronal activities in situ with spatial resolution sufficient to detail subcellular localization, (2) integrating resultant images in mass spectrometry to optically defined cell anatomy, and (3) assembling a stack of ion maps derived from mass spectra for cluster analysis. We propose that STCM IMS is the choice over a probe-based scanning mass spectrometer for high-resolution single-cell molecular imaging.<\/jats:p>","DOI":"10.3390\/cells11213382","type":"journal-article","created":{"date-parts":[[2022,10,26]],"date-time":"2022-10-26T09:59:37Z","timestamp":1666778377000},"page":"3382","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":7,"title":["Space and Time Coherent Mapping for Subcellular Resolution of Imaging Mass Spectrometry"],"prefix":"10.3390","volume":"11","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-4206-6588","authenticated-orcid":false,"given":"Jun","family":"Aoki","sequence":"first","affiliation":[{"name":"Center for Biosystems Dynamics Research, RIKEN, Saitama 351-0198, Japan"},{"name":"Graduate School of Frontier Biosciences, Osaka University, Osaka 565-0871, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0883-6112","authenticated-orcid":false,"given":"Masako","family":"Isokawa","sequence":"additional","affiliation":[{"name":"Forefront Research Center, Graduate School of Science, Osaka University, Osaka 560-0043, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5950-6039","authenticated-orcid":false,"given":"Michisato","family":"Toyoda","sequence":"additional","affiliation":[{"name":"Forefront Research Center, Graduate School of Science, Osaka University, Osaka 560-0043, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2022,10,26]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"788","DOI":"10.1038\/s41592-021-01182-8","article-title":"Metabolomic profiling of single enlarged lysosomes","volume":"18","author":"Zhu","year":"2021","journal-title":"Nat. Methods"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"1233","DOI":"10.1038\/s41592-021-01277-2","article-title":"Image-guided MALDI mass spectrometry for high-throughput single-organelle characterization","volume":"18","author":"Castro","year":"2021","journal-title":"Nat. Methods"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"5339","DOI":"10.1021\/ac049692q","article-title":"High-spatial resolution mass spectrometric imaging of peptide and protein distributions on a surface","volume":"76","author":"Luxembourg","year":"2004","journal-title":"Anal. Chem."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"57","DOI":"10.5702\/massspec.11-20","article-title":"Novel Ion Extraction Method for Imaging Mass Spectrometry","volume":"59","author":"Aoki","year":"2011","journal-title":"J. Mass Spectrom. Soc. Jpn."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"053706","DOI":"10.1063\/5.0037370","article-title":"Development of novel projection-type imaging mass spectrometer","volume":"92","author":"Aoki","year":"2021","journal-title":"Rev. Sci. Instrum."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"1229","DOI":"10.2116\/analsci.26.1229","article-title":"Functionalized manoparticles and nanostructured surfaces for surface-assisted laser desorption\/ionization mass spectrometry","volume":"26","author":"Arakawa","year":"2010","journal-title":"Anal. Sci."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"816","DOI":"10.1038\/216816a0","article-title":"Sperm maturation in rabbit epididymis","volume":"216","year":"1967","journal-title":"Nature"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"103","DOI":"10.1016\/0006-8993(84)90685-1","article-title":"GABA receptors mediate cerebral vasodilation in the unanesthetized goat","volume":"321","author":"Alborch","year":"1984","journal-title":"Brain Res."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"733","DOI":"10.1093\/molehr\/2.10.733","article-title":"Effects of gamma- aminobutyric acid on human sperm motility and hyperactivation","volume":"2","author":"Calogero","year":"1996","journal-title":"Mol. Hum. Reprod."},{"key":"ref_10","first-page":"435","article-title":"The occurrence of GABA in vas deferens, prostate, epididymis, seminal vesicle and testicle of the rat","volume":"34","author":"Erdo","year":"1983","journal-title":"Acta Biol. Hum."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"2848","DOI":"10.1523\/JNEUROSCI.06-10-02848.1986","article-title":"Evidence for GABA as a neurotransmitter in the leech","volume":"10","author":"Cline","year":"1986","journal-title":"J. Neurosci."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"967","DOI":"10.1007\/BF00970770","article-title":"Regional distribution of dopamine, 5-hydroxytryptamine, and noradrenaline in the rat vas deferens","volume":"13","author":"Celuch","year":"1988","journal-title":"Neurochem. Res."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"13","DOI":"10.1095\/biolreprod.108.068346","article-title":"Stimulating effects of dopamine on chloride transport across the rat caudal epididymal epithelium in culture","volume":"80","author":"Du","year":"2009","journal-title":"Biol. Reprod."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"2618","DOI":"10.1523\/JNEUROSCI.07-09-02618.1987","article-title":"Morphological changes in leech Retzius neurons after target contact during embryogenesis","volume":"7","author":"Jellies","year":"1987","journal-title":"J. Neurosci."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"657","DOI":"10.1111\/j.1469-7793.1999.0657n.x","article-title":"Serotonin via 5-HT1B and 5-HT2B receptors stimulates anion secretion in the rat epididymal epithelium","volume":"519","author":"Leung","year":"1999","journal-title":"J. Physiol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"136","DOI":"10.2164\/jandrol.106.000653","article-title":"Serotonin concentration, synthesis, cell origin, and targets in the rat caput epididymis during sexual maturation and variations associated with adult mating status: Morphological and biochemical studies","volume":"28","author":"Queiroz","year":"2007","journal-title":"J. Androl."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"359","DOI":"10.1016\/j.isci.2019.09.036","article-title":"Detection of a High-Turnover Serotonin Circuit in the Mouse Brain Using Mass Spectrometry Imaging","volume":"20","author":"Sugiyama","year":"2019","journal-title":"iScience"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"e4633","DOI":"10.1002\/jms.4633","article-title":"Spatial segmentation and metabolite annotation involved in sperm maturation in the rat epididymis by MALDI imaging mass spectrometry","volume":"55","author":"Lagarrigue","year":"2020","journal-title":"J. Mass Spectrom."},{"key":"ref_19","doi-asserted-by":"crossref","unstructured":"Meriaux, C., Arafah, K., Tasiemski, A., Wisztorski, M., Bruand, J., Wichlacz, C.B., Desmons, A., Debois, D., Lapre, O., and Brunelle, A. (2011). Multiple changes in peptide and lipid expression associated with regeneration in the nervous system of the medicinal leech. PLoS ONE, 6.","DOI":"10.1371\/journal.pone.0018359"},{"key":"ref_20","first-page":"2426398","article-title":"Lipidomic analysis of endocannabinoid signaling: Targeted metabolic identification and quantification","volume":"13","author":"Keereetaweep","year":"2016","journal-title":"Neural Plast."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"161","DOI":"10.1016\/j.jchromb.2011.06.025","article-title":"Simultaneous UPLC-MS quantification of the endocannabinoid 2-arachidonoylglycerol (2AG), 1-arachidonoylglycerol (1AG), and anandamide in human plasma: Minimization of matrix effects, 2AG\/1AG isomerization, and degradation by toluene solvent extraction","volume":"883","author":"Zoerner","year":"2012","journal-title":"J. Chromatogr. B"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"1001","DOI":"10.1113\/jphysiol.2005.094219","article-title":"Retrograde endocannabinoid regulation of GABAergic inhibition in the rat dentate gyrus granule cell","volume":"567","author":"Isokawa","year":"2005","journal-title":"J. Physiol."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"2766","DOI":"10.1152\/jn.00491.2010","article-title":"Endocannabinoid-dependent LTD in a nociceptive synapse requires activation of a presynaptic TRPV-like receptor","volume":"104","author":"Yuan","year":"2010","journal-title":"J. Neurophysiol."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"773","DOI":"10.1038\/42015","article-title":"A second endogenous cannabinoid that modulates long-term potentiation","volume":"388","author":"Stella","year":"1997","journal-title":"Nature"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"757","DOI":"10.1111\/j.1469-7793.2001.t01-1-00757.x","article-title":"Photolysis-induced suppression of inhibition in rat hippocampal CA1 pyramidal neurons","volume":"533","author":"Wang","year":"2001","journal-title":"J. Physiol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"67","DOI":"10.1002\/jemt.1070300106","article-title":"Epididymal epithelium: Its contribution to the formation of a luminal fluid microenvironment","volume":"30","author":"Hinton","year":"1995","journal-title":"Microsc. Res. Techn."},{"key":"ref_27","first-page":"203","article-title":"Differential distribution of GABA and GAT1 in mouse epididymis","volume":"5","author":"Qi","year":"2017","journal-title":"Annl. Clin. Labor. Res."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"451","DOI":"10.1095\/biolreprod.109.079210","article-title":"A Gradient of 2-Arachidonoylglycerol Regulates Mouse Epididymal Sperm Cell Start-Up","volume":"82","author":"Cobellis","year":"2010","journal-title":"Biol. Reprod."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"320","DOI":"10.1016\/j.ygcen.2007.02.003","article-title":"Endocannabinoid control of sperm motility: The role of epididymis","volume":"153","author":"Ricci","year":"2007","journal-title":"Gen. Comp. Endocrinol."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"12866","DOI":"10.1038\/s41598-019-49177-y","article-title":"Characterization and localization of the endocannabinoid system components in the adult human testis","volume":"9","author":"Nielsen","year":"2019","journal-title":"Sci. Rep."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"6508","DOI":"10.1523\/JNEUROSCI.0678-08.2008","article-title":"Serotonin evokes endocannabinoid release and retrogradely suppresses excitatory synapses","volume":"28","author":"Best","year":"2008","journal-title":"J. Neurosci."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"1709","DOI":"10.1016\/0306-3623(95)00065-8","article-title":"Central and peripheral 5-HT receptors in the leech (Hirudo Medicinalis) redefined","volume":"26","author":"Leake","year":"1995","journal-title":"Gen. Pharmac."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"468","DOI":"10.1126\/science.aar7042","article-title":"Multiplexed protein maps link subcellular organization to cellular states","volume":"361","author":"Gut","year":"2018","journal-title":"Science"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"236","DOI":"10.1080\/01621459.1963.10500845","article-title":"Hierarchical grouping to optimize an objective function","volume":"58","author":"Ward","year":"1963","journal-title":"J. Am. Stat. Assoc."}],"container-title":["Cells"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2073-4409\/11\/21\/3382\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T01:03:21Z","timestamp":1760144601000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2073-4409\/11\/21\/3382"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,10,26]]},"references-count":34,"journal-issue":{"issue":"21","published-online":{"date-parts":[[2022,11]]}},"alternative-id":["cells11213382"],"URL":"https:\/\/doi.org\/10.3390\/cells11213382","relation":{},"ISSN":["2073-4409"],"issn-type":[{"value":"2073-4409","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,10,26]]}}}