Cancer cell spheroids(CCS) are a valuable three-dimensional cell model in cancer studies because they could replicate numerous characteristics of solid tumors. Increasing researches have used matrix-assisted laser des...Cancer cell spheroids(CCS) are a valuable three-dimensional cell model in cancer studies because they could replicate numerous characteristics of solid tumors. Increasing researches have used matrix-assisted laser desorption/ionization mass spectrometry imaging(MALDI-MSI) to investigate the spatial distribution of endogenous compounds(e.g., lipids) in CCS. However, only limited lipid species can be detected owing to a low ion yield by using MALDI. Besides, it is still challenging to fully characterize the structural diversity of lipids due to the existence of isomeric/isobaric species. Here, we carried out the initial application of MALDI coupled with laser-postionization(MALDI-2) and trapped ion mobility spectrometry(TIMS) imaging in HCT116 colon CCS to address these challenges. We demonstrated that MALDI-2 is capable of detecting more number and classes of lipids in HCT116 colon CCS with higher signal intensities than MALDI. TIMS could successfully separate numerous isobaric/isomeric species of lipids in CCS. Interestingly, we found that some isomeric/isobaric species have totally different spatial distributions in colon CCS. Further MS/MS imaging analysis was employed to determine the compositions of fatty acid chains for isomeric species by examining disparities in signal intensities and spatial distributions of product ions. This work stresses the robust ability of TIMS and MALDI-2 imaging in analyzing endogenous lipids in CCS, which could potentially become powerful tools for future cancer studies.展开更多
Seed germination plays a pivotal role in plant growth and undergoes many intricate biochemical changes including lipid metabolism.Nevertheless,little is known about lipid changes and distributions in different structu...Seed germination plays a pivotal role in plant growth and undergoes many intricate biochemical changes including lipid metabolism.Nevertheless,little is known about lipid changes and distributions in different structures of soybean seeds during germination.Here,we applied mass spectrometry imaging(MSI)in conjunction with MS-based lipidomics to examine the lipid alterations in the embryo and cotyledon of soybean seeds during germination.To expand the coverage of lipid detection in soybean seeds,we used the novel techniques of matrix-assisted laser desorption/ionization(MALDI)and MALDI coupled with laser-postionization(MALDI-2).The results revealed that compared to MALDI,MALDI-2 enhanced the detected numbers and intensities of lipid species in various lipid classes,except for a few classes(e.g.,sphingomyelin and phosphatidylcholine).Lipidomic data showed that compared to the embryo,the cotyledon demonstrated slower but similar lipid changes during germination.These changes included the reduced levels of glycerolipids,phospholipids,and sterols,as well as the increased levels of lysophospholipids.Data from MALDI&MALDI-2 MSI supported and complemented these lipidomic findings.Our work highlights the significance of integrating lipid profiles and distributions to enhance our understanding of the metabolic pathways involved in seed germination.展开更多
基金supported by the National Natural Science Foundation of China (Nos.22036001, 22276034 and 22106130)。
文摘Cancer cell spheroids(CCS) are a valuable three-dimensional cell model in cancer studies because they could replicate numerous characteristics of solid tumors. Increasing researches have used matrix-assisted laser desorption/ionization mass spectrometry imaging(MALDI-MSI) to investigate the spatial distribution of endogenous compounds(e.g., lipids) in CCS. However, only limited lipid species can be detected owing to a low ion yield by using MALDI. Besides, it is still challenging to fully characterize the structural diversity of lipids due to the existence of isomeric/isobaric species. Here, we carried out the initial application of MALDI coupled with laser-postionization(MALDI-2) and trapped ion mobility spectrometry(TIMS) imaging in HCT116 colon CCS to address these challenges. We demonstrated that MALDI-2 is capable of detecting more number and classes of lipids in HCT116 colon CCS with higher signal intensities than MALDI. TIMS could successfully separate numerous isobaric/isomeric species of lipids in CCS. Interestingly, we found that some isomeric/isobaric species have totally different spatial distributions in colon CCS. Further MS/MS imaging analysis was employed to determine the compositions of fatty acid chains for isomeric species by examining disparities in signal intensities and spatial distributions of product ions. This work stresses the robust ability of TIMS and MALDI-2 imaging in analyzing endogenous lipids in CCS, which could potentially become powerful tools for future cancer studies.
基金supported by National Natural Science Foundation of China(No.22036001)。
文摘Seed germination plays a pivotal role in plant growth and undergoes many intricate biochemical changes including lipid metabolism.Nevertheless,little is known about lipid changes and distributions in different structures of soybean seeds during germination.Here,we applied mass spectrometry imaging(MSI)in conjunction with MS-based lipidomics to examine the lipid alterations in the embryo and cotyledon of soybean seeds during germination.To expand the coverage of lipid detection in soybean seeds,we used the novel techniques of matrix-assisted laser desorption/ionization(MALDI)and MALDI coupled with laser-postionization(MALDI-2).The results revealed that compared to MALDI,MALDI-2 enhanced the detected numbers and intensities of lipid species in various lipid classes,except for a few classes(e.g.,sphingomyelin and phosphatidylcholine).Lipidomic data showed that compared to the embryo,the cotyledon demonstrated slower but similar lipid changes during germination.These changes included the reduced levels of glycerolipids,phospholipids,and sterols,as well as the increased levels of lysophospholipids.Data from MALDI&MALDI-2 MSI supported and complemented these lipidomic findings.Our work highlights the significance of integrating lipid profiles and distributions to enhance our understanding of the metabolic pathways involved in seed germination.
