Chapter 13 Imaging of Cells and Tissues with Mass Spectrometry

Zimmerman, Tyler A.; Monroe, Eric B.; Tucker, Kevin R.; Rubakhin, Stanislav S.; Sweedler, Jonathan V.

doi:10.1016/s0091-679x(08)00613-4

Cited by 37 publications

(27 citation statements)

References 84 publications

(112 reference statements)

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“…14 There are a number of ways to apply MALDI matrix onto the sample, either by printing, 9 spraying, 5 or sublimation. 15 Each of these techniques has a number of advantages but represent a compromise between spatial resolution and analyte extraction.…”

Section: Introductionmentioning

confidence: 99%

The modified-bead stretched sample method: Development and application to MALDI-MS imaging of protein localization in the spinal cord

et al. 2011

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Matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI-MSI) has been used to create spatial distribution maps from lipids, peptides, and proteins in a variety of biological tissues. MALDI-MSI often involves trade-offs between the extent of analyte extraction and desired spatial resolution, compromises that can adversely affect detectability. For example, increasing the extraction time can lead to unwanted analyte spatial redistribution. With the stretched sample method (SSM), the extraction period can be extended, resulting in reduced analyte redistribution while suppressing detection of cationic salt adducts. The SSM involves thaw-mounting a thin tissue section onto a substrate of small glass beads embedded in Parafilm M and then stretching the membrane to fragment the tissue into thousands of bead-sized pieces. Here, we applied the SSM method to MALDI-MSI using rat spinal cord as a model. We used surface-modified beads coated with trypsin or chymotrypsin in order to facilitate controlled digestion and detection of proteins. The enzymatic reactions were maintained by repeatedly condensing water on the stretched sample surface. As a result, new peptides formed by tryptic or chymotryptic protein digestion were detected and identified using a combination of MALDI-MSI and offline liquid chromatography tandem mass spectrometric analysis. Localization of these peptides indicated the distribution of their proteins of origin, including myelin basic protein, actin beta, and tubulin alpha chain. Additionally, we used uncoated beads to create distribution maps of many endogenous lipids and small peptides. The extension of the SSM using modified beads resulted in the creation of mosaic bead surfaces where adjacent beads were coated with different enzymes or other reactive chemicals, permitting investigation of the distributions of a wider range of analytes in biological samples within a single experiment.

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Section: Introductionmentioning

confidence: 99%

The modified-bead stretched sample method: Development and application to MALDI-MS imaging of protein localization in the spinal cord

et al. 2011

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“…This approach has been successfully applied to determine the distribution pattern of neuropeptides within nervous tissue, to discover that phosphatidylcholine species increase in the brain following ischemic insult, and to determine that triacylglycerol biosynthesis varies across the tissues of a cotton embryo. [45–47] While MALDI has been effective for imaging peptides and lipids, matrix interference in the low-mass region has complicated its application to the measurement of some metabolites. [48] To image metabolites in the low-mass region a technology has emerged known as nanostructure-imaging mass spectrometry (NIMS, also referred to as nanostructure-initiator mass spectrometry).…”

Section: In Situ Imaging Of Metabolitesmentioning

confidence: 99%

After the feature presentation: technologies bridging untargeted metabolomics and biology

Cho

Mahieu

Johnson

et al. 2014

Current Opinion in Biotechnology

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Liquid chromatography/mass spectrometry-based untargeted metabolomics is now an established experimental approach that is being broadly applied by many laboratories worldwide. Interpreting untargeted metabolomic data, however, remains a challenge and limits the translation of results into biologically relevant conclusions. Here we review emerging technologies that can be applied after untargeted profiling to extend biological interpretation of metabolomic data. These technologies include advances in bioinformatic software that enable identification of isotopes and adducts, comprehensive pathway mapping, deconvolution of MS2 data, and tracking of isotopically labeled compounds. There are also opportunities to gain additional biological insight by complementing the metabolomic analysis of homogenized samples with recently developed technologies for metabolite imaging of intact tissues. To maximize the value of these emerging technologies, a unified workflow is discussed that builds on the traditional untargeted metabolomic pipeline. Particularly when integrated together, the combination of the advances highlighted in this review helps transform lists of masses and fold changes characteristic of untargeted profiling results into structures, absolute concentrations, pathway fluxes, and localization patterns that are typically needed to understand biology.

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“…These molecular signatures are typically comprised of 5–20 different proteins that together result in robust diagnostic patterns. 21, 22 IMS-based studies have been used to elucidate molecular signatures of different tumor types and grades including brain, oral, lung, breast, gastric, pancreatic, renal, ovarian and prostate cancer. 23–26 …”

Section: Introductionmentioning

confidence: 99%

Imaging Mass Spectrometry—A New and Promising Method to Differentiate Spitz Nevi From Spitzoid Malignant Melanomas

Lazova

Seeley²,

Keenan³

et al. 2012

The American Journal of Dermatopathology

105

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Background Differentiating Spitz nevus (SN) from Spitzoid malignant melanoma (SMM) is one the most difficult problems in Dermatopathology. Specific Aim To identify differences on proteomic level between SN and SMM. Methods We performed Imaging Mass Spectrometry (IMS) analysis on formalin-fixed, paraffin embedded tissue samples (FFPE) to identify differences on proteomic level between SN and SMM. The diagnosis of SN and SMM was based on histopathologic criteria, clinical features, and follow up data, which confirmed that none of the lesions diagnosed as SN recurred or metastasized. The melanocytic component (tumor) and tumor microenvironment (dermis) from 114 cases of SN and SMM from the Yale Spitzoid Neoplasm Repository were analyzed. After obtaining mass spectra from each sample, classification models were built using a training set of biopsies from 26 SN and 25 SMM separately for tumor and for dermis. The classification algorithms developed on the training data set were validated on another set of 30 samples from SN and 33 from SMM. Results We found proteomic differences between the melanocytic components of SN and SMM and identified five peptides that were differentially expressed in the two groups. From these data, 29 out of 30 SN and 26 out of 29 SMM were recognized correctly based on tumor analysis in the validation set. This method correctly classified SN with 97% sensitivity and 90% specificity in the validation cohort. Conclusions IMS analysis can reliably differentiate SN from SMM in FFPE tissue based on proteomic differences.

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Chapter 13 Imaging of Cells and Tissues with Mass Spectrometry

Cited by 37 publications

References 84 publications

The modified-bead stretched sample method: Development and application to MALDI-MS imaging of protein localization in the spinal cord

The modified-bead stretched sample method: Development and application to MALDI-MS imaging of protein localization in the spinal cord

After the feature presentation: technologies bridging untargeted metabolomics and biology

Imaging Mass Spectrometry—A New and Promising Method to Differentiate Spitz Nevi From Spitzoid Malignant Melanomas

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