Advances in cellular, subcellular, and nanoscale imaging in vitro and in vivo

Wessels, Johannes T.; Yamauchi, Kensuke; Hoffman, Robert M.; Wouters, Fred S.

doi:10.1002/cyto.a.20931

Cited by 56 publications

(41 citation statements)

References 56 publications

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“…While this approach has been valuable for compound identification and quantification, this methodology is limited in its ability to distinguish where these molecules were localized in situ. On the other hand, optical imaging techniques, such as confocal or electron microscopy using selected fluorescent dyes, antibodies, and chemical modifications, can hint at cellular localization (Eggeling et al, 2009;Wessels et al, 2010) but currently cannot derive comprehensive compositional information within these images alone.…”

Section: Introductionmentioning

confidence: 99%

Spatial Mapping of Lipids at Cellular Resolution in Embryos of Cotton

et al. 2012

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Advances in mass spectrometry (MS) have made comprehensive lipidomics analysis of complex tissues relatively commonplace. These compositional analyses, although able to resolve hundreds of molecular species of lipids in single extracts, lose the original cellular context from which these lipids are derived. Recently, high-resolution MS of individual lipid droplets from seed tissues indicated organelle-to-organelle variation in lipid composition, suggesting that heterogeneity of lipid distributions at the cellular level may be prevalent. Here, we employed matrix-assisted laser desorption/ ionization-MS imaging (MALDI-MSI) approaches to visualize lipid species directly in seed tissues of upland cotton (Gossypium hirsutum). MS imaging of cryosections of mature cotton embryos revealed a distinct, heterogeneous distribution of molecular species of triacylglycerols and phosphatidylcholines, the major storage and membrane lipid classes in cotton embryos. Other lipids were imaged, including phosphatidylethanolamines, phosphatidic acids, sterols, and gossypol, indicating the broad range of metabolites and applications for this chemical visualization approach. We conclude that comprehensive lipidomics images generated by MALDI-MSI report accurate, relative amounts of lipid species in plant tissues and reveal previously unseen differences in spatial distributions providing for a new level of understanding in cellular biochemistry.

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Section: Introductionmentioning

confidence: 99%

Spatial Mapping of Lipids at Cellular Resolution in Embryos of Cotton

et al. 2012

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“…Fluorescent proteins with various emission wavelengths, including the green fluorescent protein (GFP) and red-emitting fluorescent proteins (RFP), have been used for in vivo imaging [33]. The high flexibility GFP provides a permanent and heritable label in living cells.…”

Section: Mirna Imaging Analysismentioning

confidence: 99%

Intracellular and Organic miRNA In Situ Detection

Zhang

Dong

Tian

2015

SpringerBriefs in Molecular Science

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The detection methods of miRNA in intracellular or organisms fall into two broad categories: indirect detection and direct analysis. The indirect measurement of the expression levels of miRNAs in cells and tissues involves cells lysis and detection by qRT-PCR, northern blotting, or microarray hybridization. The direct analysis methods are a noninvasive manner for repetitively monitoring and obtaining real-time imaging of the intracellular miRNA by using imaging analysis or in situ hybridization (ISH). Technologies for direct detection of the temporal and spatial expression sequence of specific miRNA in cells or tissues are extremely important for elucidating miRNA biology. The progress of optical imaging techniques with multimodal reporter systems holds great promise for noninvasive and real-time imaging of molecular agent expression in living cell. Recent progress in nanotechnology and imaging detection techniques leads to multifunctional nanoprobe with specific-transfection, tracing, and regulation function in intracellular miRNA detection. ISH holds great promise for visualization of the spatial localization of RNA at the tissue, cellular, and even subcellular level.Keywords Intracellular miRNA · Organic miRNA · In situ detection · Cell imaging analysis · Functional nanoprobeThe detection of intracellular miRNA is significant in the development of gene therapy and gene medicines. The deficiency of small size and degradation of the mature miRNAs make it difficult to directly transfer the specific miRNA into the cell. A noninvasive manner for repetitively monitoring and obtaining real-time imaging of the intracellular miRNA is required for the analysis of miRNAs in practical clinic application. Efficient gene vectors, including viral [1] and nonviral categories, [2] were usually required to translocate miRNA probe through membrane barrier into the cell. Retrovirus and adenovirus vectors can be effectively transferred via the gene probes (DNA, RNA) into most cell lines. However, the

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“…[1][2][3][4][5] Organic dyes, [6][7][8][9][10] engineered fluorescent proteins [11][12][13][14] and quantum dots [15][16][17][18] are extensively used for imaging and sensing of concerned biological targets. However, intrinsic limitations of these fluorescent probes restrict their applications, such as fast photobleaching of organic dyes and fluorescent proteins, [19][20][21] heavy-metal core-related cytotoxicity and difficulties in surface modification of quantum dots.…”

Section: Introductionmentioning

confidence: 99%