An examination of specific cellular organelle-targeting nanotags using combined 3D Raman and SERS imaging
Poster Apr 08, 2015
Katherine Lau, Sarah McAughtrie, Karen Faulds, Duncan Graham
Metal nanoparticles (NPs), such as silver/gold NPs, are ideal for enhancing cell imaging sensitivity. Through their surface plasmon and electric field effects, NPs can greatly enhance the Raman signals of the adsorbed molecules. This effect is termed surface enhanced Raman scattering (SERS). When internalised by cells, NPs can enhance the cells’ intrinsic Raman signals . By tagging NPs with reporter molecules, nanotags can be delivered into cells for cellular imaging, and be distinguished through their reporters’ SERS signatures . This capability benefits research which involves the intracellular delivery of NPs, e.g. thermal therapy of cancer and transporting drug molecules into cells.
We have previously demonstrated the use of combined 3D SERS and Raman imaging for the verification of intracellular nanotag delivery. By using a Renishaw inVia Raman microscope coupled to 532 nm and 633 nm laser sources, both the cells’ intrinsic Raman signals and the NPs’ SERS signals were recorded. By overlaying the datasets, we successfully revealed the locations of the nanotags within the cells in a three dimensional manner .
Specific targeting of cellular organelles by NPs can be achieved by functionalization with peptide targets. In this work, we investigated the specific targeting of endoplasmic reticulum and trans-Golgi network in Chinese hamster ovarian cells using functionalised nanotags. The targeting was further examined using the combined 3D SERS and Raman imaging method.
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PfABCk2 gene is ligated into pET21a+ vector with His-tag at C-terminus and transformed into BL-21 (DE3) competent cells that are verified through Miniprep and DNA sequencing. Furthermore, this gene construct is utilized to heterologous express this protein with IPTG and afterwards purified the recombinant protein kinase using nickel affinity chromatography as shown on 10% SDS-PAGE with the expected 36 kDa protein band.READ MORE
Novel Role of the Innate Immune DNA Sensor IFI16 (Interferon Gamma Inducible Protein 16) as a Major Epigenetic Modulator During KSHV Infection and Lytic ReactivationPoster
Studies have shown that IFI16 acts as an antiviral restriction factor against a number of DNA viruses, by inhibiting viral replication or transcription through epigenetic modifications. However, till date, no specific epigenetic function of IFI16 has been identified. Here, we have discovered that IFI16 recruits two histone methyltransferases on the KSHV episome leading to altered Histone H3K9 methylation, thus regulating its lifecycle.READ MORE
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9th International Conference on Mass Spectrometry and Chromatography
Sep 21 - Sep 22, 2018