The use of highly sensitive cameras turns camera-based confocal systems into the ideal solution to capture all signals from expanded samples, even the dimmest ones. Researchers need to use a microscope that can facilitate optimal delivery of light to the samples and extremely efficient capture of all the photons from the sample, i.e. To overcome the loss of signal in expanded samples, a highly sensitive acquisition system is needed. Technology Solutions for Imaging Expanded Samples These requirements are problematic and difficult to achieve with conventional microscopes. Other challenges consist of dealing with the expansion itself including imaging a very large field of view (Figure 1), and imaging deep into the sample. Therefore, imaging expanded samples requires an instrument that is sensitive enough to detect low light signals. During the actual expansion, a 4-fold expansion means a 64-fold volumetric expansion hence the fluorophore density and brightness of the fluorescence signal is lowered by 64x. However, the fluorescence signal can decrease up to 60% during homogenisation. Next, the homogenisation step aims to avoid sample distortion and ensure that the specimen organisation is kept intact. During gelation, the inefficient crosslinking results in fragmented signal preservation in the hydrogel. During the expansion procedure monomers of a polymer are delivered into the cells, and then, once inside, the polymerisation is triggered (gelation). The complexity of the fixation procedure applied to the organelles causes problems in antibody labelling, leading to samples with a low signal.
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One of the main challenges is the loss of fluorescence induced by the different steps of the protocol. Nevertheless, there are challenges to overcome to successfully image expanded samples.
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For a full summary of the protocols of expansion microscopy, please read our technical article – What is Expansion Microscopy? This technique has been gaining a lot of attention not only because it is an alternative approach to super-resolution, but also because it can be combined with current super-resolution techniques and increase even further the limits of what the microscopes “can see”. Instead of increasing the optical resolution of the microscope, Expansion Microscopy “expands” the sample isotopically.
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An alternative approach to super-resolution methods was developed by Edward Boyden’s laboratory at MIT in 2015. Police detained several people Monday during scuffles between Saakashvili’s supporters and officers.The resolution in light microscopy is limited by the diffraction limit of light, which means that structures that are closer than 200 nm apart cannot be distinguished unless researchers use Super-resolution techniques.
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Saakashvili is also facing charges related to the raiding of the Imedi TV company and the seizure of the property of its owner, the late businessman Badri Patarkatsishvili. “The whole world is amazed that I am addressing you through this glass wall instead of walking freely in the country I have built,” the former president said during the court hearing. Speaking in court, Saakashvili said he doesn’t recognize the authority of the prosecutor’s office and used the hearings to denounce the Georgian authorities. Monday’s hearings at the Tbilisi City Court on a new case against Saakashvili for the violent dispersal of a 2007 protest marked Saakashvili’s first public appearance since his arrest. His lawyers and doctors said it caused him neurological damage. 19 after authorities agreed to transfer him from a prison clinic to a military hospital. Saakashvili went on a seven-week hunger strike following his arrest, ending it on Nov. 1 after returning to Georgia to try to bolster opposition forces before nationwide municipal elections.
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Mikheil Saakashvili, who served as president in 2008-13, left Georgia for Ukraine after the end of his second term and was later convicted in absentia of abuse of power and sentenced to six years in prison. TBILISI, Georgia (AP) - The imprisoned former president of ex-Soviet republic Georgia appeared in court Monday on charges of abuse of office, proceedings that triggered scuffles between his supporters and police.