Molecular Histology Analysis of Cryopreserved Tissue Using Peptide/Protein MALDI-TOF Imaging Mass Spectrometry (MALDI-IMS)

2021 ◽  
pp. 177-190
Author(s):  
Ibon Iloro ◽  
Iraide Escobés ◽  
Mikel Azkargorta ◽  
Félix Elortza
Keyword(s):  
Mass Spectrometry ◽  
Imaging Mass Spectrometry ◽  
Maldi Tof ◽  
Maldi Ims

Methods for Human Olfactory Bulb Tissue Studies Using Peptide/Protein MALDI-TOF Imaging Mass Spectrometry (MALDI-IMS)

2017 ◽  
pp. 91-106 ◽  
Author(s):  
Ibon Iloro ◽  
Joaquín Fernández-Irigoyen ◽  
Iraide Escobes ◽  
Mikel Azkargorta ◽  
Enrique Santamaría ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Olfactory Bulb ◽  
Imaging Mass Spectrometry ◽  
Maldi Tof ◽  
Maldi Ims

scholarly journals High-resolution imaging mass spectrometry combined with transcriptomic analysis identified a link between fatty acid composition of phosphatidylinositols and the immune checkpoint pathway at the primary tumour site of breast cancer

2019 ◽  
Vol 122 (2) ◽  
pp. 245-257 ◽  
Author(s):  
Masahiro Kawashima ◽  
Mariko Tokiwa ◽  
Tomomi Nishimura ◽  
Yukiko Kawata ◽  
Masahiro Sugimoto ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Mass Spectrometry ◽  
Fatty Acid ◽  
Cancer Cells ◽  
Immune Checkpoint ◽  
Imaging Mass Spectrometry ◽  
Invasive Cancer ◽  
Fatty Acid Binding ◽  
Checkpoint Pathway ◽  
Maldi Ims

Abstract Background The fatty acid (FA) composition of phosphatidylinositols (PIs) is tightly regulated in mammalian tissue since its disruption impairs normal cellular functions. We previously found its significant alteration in breast cancer by using matrix-assisted laser desorption and ionisation imaging mass spectrometry (MALDI-IMS). Methods We visualised the histological distribution of PIs containing different FAs in 65 primary breast cancer tissues using MALDI-IMS and investigated its association with clinicopathological features and gene expression profiles. Results Normal ductal cells (n = 7) predominantly accumulated a PI containing polyunsaturated FA (PI-PUFA), PI(18:0/20:4). PI(18:0/20:4) was replaced by PIs containing monounsaturated FA (PIs-MUFA) in all non-invasive cancer cells (n = 12). While 54% of invasive cancer cells (n = 27) also accumulated PIs-MUFA, 46% of invasive cancer cells (n = 23) accumulated the PIs-PUFA, PI(18:0/20:3) and PI(18:0/20:4). The accumulation of PI(18:0/20:3) was associated with higher incidence of lymph node metastasis and activation of the PD-1-related immune checkpoint pathway. Fatty acid-binding protein 7 was identified as a putative molecule controlling PI composition. Conclusions MALDI-IMS identified PI composition associated with invasion and nodal metastasis of breast cancer. The accumulation of PI(18:0/20:3) could affect the PD-1-related immune checkpoint pathway, although its precise mechanism should be further validated.

scholarly journals Lipidoak aztertu eta identifikatzeko TLC kromatografiaren eta MALDI masa-espektrometriaren arteko akoplamendua

2018 ◽  
pp. 149-165
Author(s):  
Antonio Veloso
Keyword(s):  
Mass Spectrometry ◽  
Imaging Mass Spectrometry ◽  
Laser Desorption ◽  
Laser Desorption Ionization ◽  
Ionization Time ◽  
Flight Mass Spectrometry ◽  
Maldi Ims ◽  
Layer Chromatography ◽  
Matrix Assisted Laser ◽  
Matrix Assisted Laser Desorption

Geruza meheko kromatografia teknika (TLC, ingelesezko Thin Layer Chromatography) oso erabilia izan da hainbat molekularen banaketa eta karakterizaziorako. Bestalde, Matrix Assisted Laser Desorption Ionization Time of Flight Mass Spectrometry (MALDI-TOF MS) edo matrizez lagundutako laserraren bidezko desortzioionizazioa/hegaldi-denbora masa-espektrometria teknikaren erabilera gero eta gehiago hedatzen ari da molekula desberdinen analisia egiteko. Gainera, Imaging Mass Spectrometry (IMS) edo masa-espektrometriaren bidezko irudia izeneko teknikak, ahalbidetzen du ehun biologikoetan dauden lipidoen, proteinen eta farmakoen banaketa-irudiak lortzea eta teknika bikaina da molekula horien banaketa aztertzeko. Teknika honek lipido mota ezberdinen identifikazioa eta lokalizazioa ahalbidetzen du, konposatuaren edota familia kimikoaren aukera egin aurretik. Analizatu behar diren konposatuen aukera aldez aurretik egin behar ez denez, aukera dago irudiak sortzeko masa-espektrometroan detektatutako ioietatik abiatuta. Lan honetan, erakusten da lipidoen ikerkuntzarako oso lagungarria dela TLC xaflak eta MALDI IMS teknikak konbinatzea.

scholarly journals Tissue Localization of Glycosphingolipid Accumulation in a Gaucher Disease Mouse Brain by LC-ESI-MS/MS and High-Resolution MALDI Imaging Mass Spectrometry

2017 ◽  
Vol 22 (10) ◽  
pp. 1218-1228 ◽  
Author(s):  
E. Ellen Jones ◽  
Wujuan Zhang ◽  
Xueheng Zhao ◽  
Cristine Quiason ◽  
Stephanie Dale ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Gaucher Disease ◽  
Imaging Mass Spectrometry ◽  
Maldi Imaging ◽  
Differential Signal ◽  
Esi Ms ◽  
Maldi Ims ◽  
Imaging Results ◽  
Hematoxylin And Eosin ◽  
And Control

To better understand regional brain glycosphingolipid (GSL) accumulation in Gaucher disease (GD) and its relationship to neuropathology, a feasibility study using mass spectrometry and immunohistochemistry was conducted using brains derived from a GD mouse model (4L/PS/NA) homozygous for a mutant GCase (V394L [4L]) and expressing a prosaposin hypomorphic (PS-NA) transgene. Whole brains from GD and control animals were collected using one hemisphere for MALDI FTICR IMS analysis and the other for quantitation by LC-ESI-MS/MS. MALDI IMS detected several HexCers across the brains. Comparison with the brain hematoxylin and eosin (H&E) revealed differential signal distributions in the midbrain, brain stem, and CB of the GD brain versus the control. Quantitation of serial brain sections with LC-ESI-MS/MS supported the imaging results, finding the overall HexCer levels in the 4L/PS-NA brains to be four times higher than the control. LC-ESI-MS/MS also confirmed that the elevated hexosyl isomers were glucosylceramides rather than galactosylceramides. MALDI imaging also detected differential analyte distributions of lactosylceramide species and gangliosides in the 4L/PS-NA brain, which was validated by LC-ESI-MS/MS. Immunohistochemistry revealed regional inflammation, altered autophagy, and defective protein degradation correlating with regions of GSL accumulation, suggesting that specific GSLs may have distinct neuropathological effects.

scholarly journals Mass spectrometric imaging of metabolites in kidney tissues from rats treated with furosemide

2016 ◽  
Vol 310 (11) ◽  
pp. F1317-F1327 ◽  
Author(s):  
Jin Woo Jung ◽  
Mi Suk Lee ◽  
Hyo-Jung Choi ◽  
Sunhee Jung ◽  
Yu-Jung Lee ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Spatial Distribution ◽  
Rat Kidney ◽  
Imaging Mass Spectrometry ◽  
Urine Osmolality ◽  
Free Access ◽  
Sprague Dawley ◽  
Fixed Amount ◽  
Maldi Tof ◽  
Relative Quantitation

In the kidney, metabolic processes are different among the cortex (COR), outer medulla (OM), and inner medulla (IM). Using matrix-assisted laser desorption/ionization (MALDI) and imaging mass spectrometry (IMS), we examined the change of metabolites in the COR, OM, and IM of the rat kidney after furosemide treatment compared with vehicle-treated controls. Osmotic minipumps were implanted in male Sprague-Dawley rats to deliver 12 mg·day−1·rat−1 of furosemide. Vehicle-treated ( n = 14) and furosemide-treated (furosemide rats, n = 15) rats in metabolic cages received a fixed amount of rat chow (15 g·220 g body wt−1·day−1 for each rat) with free access to water intake for 6 days. At day 6, higher urine output (32 ± 4 vs. 9 ± 1 ml/day) and lower urine osmolality (546 ± 44 vs. 1,677 ± 104 mosmol/kgH2O) were observed in furosemide rats. Extracts of COR, OM, and IM were analyzed by ultraperformance liquid chromatography coupled with quadrupole time-of-flight (TOF) mass spectrometry, where multivariate analysis revealed significant differences between the two groups. Several metabolites, including acetylcarnitine, betaine, carnitine, choline, and glycerophosphorylcholine (GPC), were significantly changed. The changes of metabolites were further identified by MALDI-TOF/TOF and IMS. Their spatial distribution and relative quantitation in the kidneys were analyzed by IMS. Carnitine compounds were increased in COR and IM, whereas carnitine and acetylcarnitine were decreased in OM. Choline compounds were increased in COR and OM but decreased in IM from furosemide rats. Betaine and GPC were decreased in OM and IM. Taken together, MALDI-TOF/TOF and IMS successfully provide the spatial distribution and relative quantitation of metabolites in the kidney.

scholarly journals Defining Changes in the Spatial Distribution and Composition of Brain Lipids in the Shiverer and Cuprizone Mouse Models of Myelin Disease

2018 ◽  
Vol 67 (3) ◽  
pp. 203-219 ◽  
Author(s):  
Rajanikanth J. Maganti ◽  
Xiaoping L. Hronowski ◽  
Robert W. Dunstan ◽  
Brian T. Wipke ◽  
Xueli Zhang ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Spatial Distribution ◽  
Mouse Model ◽  
Morphological Changes ◽  
Imaging Mass Spectrometry ◽  
Ionization Mass ◽  
Brain Lipids ◽  
Esi Ms ◽  
Maldi Ims ◽  
Lipid Species

Myelin is composed primarily of lipids and diseases affecting myelin are associated with alterations in its lipid composition. However, correlation of the spatial (in situ) distribution of lipids with the disease-associated compositional and morphological changes is not well defined. Herein we applied high resolution matrix-assisted laser desorption ionization imaging mass spectrometry (MALDI-IMS), immunohistochemistry (IHC), and liquid chromatography–electrospray ionization–mass spectrometry (LC-ESI-MS) to evaluate brain lipid alterations in the dysmyelinating shiverer (Shi) mouse and cuprizone (Cz) mouse model of reversible demyelination. MALDI-IMS revealed a decrease in the spatial distribution of sulfatide (SHexCer) species, SHexCer (d42:2), and a phosphatidylcholine (PC) species, PC (36:1), in white matter regions like corpus callosum (CC) both in the Shi mouse and Cz mouse model. Changes in these lipid species were restored albeit not entirely upon spontaneous remyelination after demyelination in the Cz mouse model. Lipid distribution changes correlated with the local morphological changes as confirmed by IHC. LC-ESI-MS analyses of CC extracts confirmed the MALDI-IMS derived reductions in SHexCer and PC species. These findings highlight the role of SHexCer and PC in preserving the normal myelin architecture and our experimental approaches provide a morphological basis to define lipid abnormalities relevant to myelin diseases.

scholarly journals Early lipid changes in acute kidney injury using SWATH lipidomics coupled with MALDI tissue imaging

2016 ◽  
Vol 310 (10) ◽  
pp. F1136-F1147 ◽  
Author(s):  
Sangeetha Rao ◽  
Kelly B. Walters ◽  
Landon Wilson ◽  
Bo Chen ◽  
Subhashini Bolisetty ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Acute Kidney Injury ◽  
Ischemia Reperfusion ◽  
Imaging Mass Spectrometry ◽  
Kidney Injury ◽  
Tissue Imaging ◽  
Maldi Ims ◽  
Hospital Morbidity ◽  
Rate Limiting

Acute kidney injury (AKI) is one of the leading causes of in-hospital morbidity and mortality, particularly in critically ill patients. Although our understanding of AKI at the molecular level remains limited due to its complex pathophysiology, recent advances in both quantitative and spatial mass spectrometric approaches offer new opportunities to assess the significance of renal metabolomic changes in AKI models. In this study, we evaluated lipid changes in early ischemia-reperfusion (IR)-related AKI in mice by using sequential window acquisition of all theoretical spectra (SWATH)-mass spectrometry (MS) lipidomics. We found a significant increase in two abundant ether-linked phospholipids following IR at 6 h postinjury, a plasmanyl choline, phosphatidylcholine (PC) O-38:1 (O-18:0, 20:1), and a plasmalogen, phosphatidylethanolamine (PE) O-42:3 (O-20:1, 22:2). Both of these lipids correlated with the severity of AKI as measured by plasma creatinine. In addition to many more renal lipid changes associated with more severe AKI, PC O-38:1 elevations were maintained at 24 h post-IR, while renal PE O-42:3 levels decreased, as were all ether PEs detected by SWATH-MS at this later time point. To further assess the significance of this early increase in PC O-38:1, we used matrix-assisted laser desorption ionization imaging mass spectrometry (MALDI-IMS) to determine that it occurred in proximal tubules, a region of the kidney that is most prone to IR injury and also rich in the rate-limiting enzymes involved in ether-linked phospholipid biosynthesis. Use of SWATH-MS lipidomics in conjunction with MALDI-IMS for lipid localization will help in elucidating the role of lipids in the pathobiology of AKI.

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