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.
Plasmodium falciparum is evolving resistance to Artemisinin Combination Therapy. The gene with the strongest association with resistance is K13. K13 is an ortholog of the well characterized transcriptional regulator Keap1. In this work we transcriptionally characterized a mutant with a transposon inserted in the K13 promoter region which results in dysregulation of K13 at 2 points of the intraerythrocytic cycle of the life-cycle to identify the processes regulated by K13.READ MORE
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