MICROSCOPY & IMAGING
be treated with cardiac resynchronisation therapy (CRT). Tis therapy can resynchronise the ventricular mechanical and electrical activity, reducing mortality in patients. SML microscopy has been used to investigate the sarcomeric organisation pre- and post-CRT treatment using α-actinin as a marker to reveal the sarcomeric structures.
FIG. 2. The synaptic labelling of C. elegans. Three markers present in this image: yellow (Skylan-s Active zone marker), magenta (HaloTag::JF646 Calcium Channel 1) and cyan (SNAPf::JF549 Calcium Channel 2). Image courtesy of Sean Merrill and Dr Erik Jorgensen, University of Utah
SML IN NEUROSCIENCE Neuroimaging tools, such as SML microscopy, can be used to answer complex questions regarding neurological and psychiatric disorders. Answers to these questions can lead to the development of innovative strategies to prevent the clinical manifestation of devastating conditions. Te high resolution and magnification
used by SML microscopy can produce images of extremely small neurological structures; clear images of these structures would be impossible with any other microscope. One such example is the use of SML microscopy to label the synapses in Caenorhabditis elegans. Tis has allowed scientists to study and understand the relationship between the membrane-bound calcium sensor and endosomes in endocytosis. Only SML can distinguish these domains, since the synaptic structures themselves are often smaller than the diffraction limit of light. SML has also been used to determine the spatial distribution of the vesicular monoamine transporter 2 (VMAT2) pre- and post-drug treatment in the rat brain tissue sections.
SML IN ONCOLOGY Cancer research and treatment has advanced greatly over the past few decades. However, it is an extremely complex and heterogenous disease, and there are many different types of tumours – many we know little about. Preclinical and clinical research on potential drugs is
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needed to determine potential toxicities, confirm the efficacy and establish any off-target effects. Imaging technologies can be employed to study the effects of treatments in preclinical models and uncover key tumour signalling and progression mechanisms. For example, SML has been used to
investigate HuH7 – a hepatocellular carcinoma cell line – using markers for the endoplasmic reticulum via an RNA binding protein that labels stress granules. Furthermore, genomic imaging is a powerful tool to study cancer biology, as structural changes within the genome are associated with the heterogeneity of cancers.
SML IN CARDIOLOGY In the USA, approximately 647,000 people die from cardiovascular disease per year. Imaging technologies are used to provide detailed insights into the cardiovascular system, driving the development of diagnostic and treatment strategies. Te high resolution and low magnification offered by SML microscopy has helped solve many complex questions within cardiology research. 40% of patients suffering from heart
failure develop delays in ventricular electrical activation, which can result in ventricular dyssynchrony – the difference in timing or synchronisation of ventricle contraction. Te downstream effect of this may be cardiac death. However, if detected, dyssynchronous heart failure can
SML IN DEVELOPMENTAL AND CELL BIOLOGY Disease modelling and the development of future treatment resources relies on the fundamental knowledge gained from exploring the inner working of cells, along with understanding the intricacies of animal and plant development. Biological imaging produced by SML microscopy can be used to track individual biomolecules or observe biological processes. Samples are labelled with organic dyes, proteins or quantum dots, which enables the SML microscope to image single fluorophores at high speed and in three dimensions. Tis process is highly advantageous for any scientist wanting to capture live biological motion, which is difficult with many other microscopes.
Tis is just a short overview of some of the many emerging applications where super resolution microscopy in general, and single-molecule localisation in particular, are helping researchers to advance their understanding of complex structures.
FIG. 3. A routine three colour experiment using the Vutara 352 SML microscope to image U2OS cells. Image is courtesy of Leremy Colf and Dr Wes Sundquist University of Utah
Lauren Gagnon, Ph.D. is an application scientist at Bruker.
www.bruker.com
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