In January 2017, Argentine and German scientists unveiled advanced microscopy technology with unprecedented resolution in Science.Called Minflux, it allowed to obtain with a fluorescence microscope (using only light and lenses) images with a resolution of one nanometer or more;that is, details 10 million times smaller than a centimeter.It was considered one of the ten most important advances of the year.However, from that moment until now, few teams have managed to use it.“In fact, as far as I know, there are only three laboratories in the world that dominate it: that of Stefan Hell [Nobel Prize in Chemistry 2014 for having managed, together with Eric Betzig and William Moerner, to circumvent the difficulties of seeing objects less than 200 or 300 nanometers (billionths of a meter) using fluorescent dyes], ours and another from Munich, who put together such a microscope with our help, says Fernando Stefani, director of the Bionanosciences Research Center (Cibion) of the Conicet, located in the Scientific Pole of the former Bodegas Giol, co-author of that work and leader of the Argentine group–.This is because it is a fairly complex technique and, although the work had many citations, it was hardly applied”.Another obstacle, not negligible, is that it requires a device of more than a million, or a million and a half dollars.But now, honoring their proverbial ingenuity, Argentine scientists have just published in Light.Science & Applications, a high-impact journal from the Nature group, a study in which they show that the same resolution can be achieved by applying similar concepts and making some modifications to a confocal microscope, such as those found in many laboratories in the country."They are 'scanning' microscopes, in the sense that a photo is not taken with a camera, but a laser focused on the sample is scanned, it is scanned point by point and the image is reconstructed," explains Stefani. .They are among the most common, the ones used by all biologists.”These devices are mainly used in biomedicine because cells, tissues, organs are formed from the self-assembly of proteins.Proteins are the building blocks of biological systems;They are arranged in different ways, one next to the other.Because they are only a few nanometers in size, just this level of resolution is needed to study the structure of biological systems."With an electron microscope you can obtain these images, but you have to prepare the sample in a way that is not compatible with life," explains the scientist.then one is always left with the doubt of whether that photo really shows what happens in a natural environment.Fluorescence microscopy allows imaging under conditions that are compatible with life.So the information you get is much more likely to be accurate.”Confocal microscopes allow three-dimensional images to be made, always with resolutions of 300 nanometers or more, because they are limited by diffraction."But what we show is that, with a couple of modifications, they can be turned into a microscope of the highest resolution," he notes.Until the 1990s, most microscopists thought that, due to limitations of physical laws, it would be impossible to see objects smaller than 200 or 300 nanometers using light.Hell, Betzig and Moerner paved the way to overcome that impediment.But although in theory his method should allow reaching a precision of one nanometer, in practice it was not achieved.It only reached 20 or 30 nanometers.With Minflux technology, it was shown that an optical microscope can achieve the highest possible resolution;that is, the very size of the molecule that emits light.Fluorophores [fluorescent substances] absorb light of one color and emit light of another color.But since they have to absorb and emit energy, sooner or later they degrade.Therefore, the fluorescence photons they can give off are limited.This method maximizes the information of each photon and allows reaching a resolution of the size of the molecules themselves.The modifications that they now propose in the new work are simple."In general, in confocal microscopes, what is swept over the sample to obtain an image is a focused laser," Stefani points out.A dot of maximum intensity is generated that is directed towards different places in the sample.What you have to do is modify the lighting beam, so that instead of focusing with a maximum intensity in the center, it has a minimum intensity in the center and maximums around it.There are many ways to do it.And the other thing (which can actually be useful for any microscope) is to have a stabilization system for the sample, so that it remains very still in the microscope.Because these measurements take several minutes and any conventional device has a miscalibration of around 10, 20 or 30 nanometers per minute.Even if you put the sample in and leave it still, it moves because of mechanical or thermal fluctuations.It may move 30 nanometers in a minute, which for any application is negligible, but in this case it becomes important because it has to be completely fixed for the entire time it is imaged.You have to move less than a nanometer to be able to take the photo you are looking for.”To test their ideas, the scientists collaborated with a Swiss group led by fellow Argentine Guillermo Acuña, who provided them with "DNA origami," consisting of DNA folded by default."It is widely used in nanotechnology and for 'super resolution' methodologies, because it allows molecules to be placed in predetermined positions at very small distances," the specialist explains.It is a technology that we handle in the laboratory, but we are not very experts.We have been working with them for years and they specially prepared one of those arrangements for us, given that to demonstrate that the technique really fulfilled what we wanted, a reference sample was needed”.Stefani emphasizes that with this system it is possible to locate in a sample where each molecule is with a precision equal to its size.“It is similar to what happens with GPS –he illustrates–.You can indicate where you are with 100 meters of error, with 10 or with one.But if the error is less than a meter, it says exactly where your head and arm are, because it's the same size as you.We can tell where each molecule is with a level of precision that is comparable to or equal to the size of the molecule itself.It doesn't make sense anymore."Although he clarifies that it is not his area of ​​specialty, the physicist and nanotechnologist Carlos Balseiro, who did not participate in this development, considers it very positive that these achievements can be obtained from the country."This is exciting news! –he is enthusiastic–. As the authors say, it is not about allowing things to be done that were impossible until now, but about being able to do them without having to depend on a very few groups in the world, something like like democratizing single molecule detection capabilities, and when techniques become widespread is when they really hit their full potential."This advance makes them more accessible, but protocols for sample preparation and data analysis will have to be developed.“The other job was a huge success, but then we were left wondering: is this really the best way to do it?Stefani sums it up.In these years, we have been investigating other options.First we advance in the theoretical-conceptual aspect, we develop a framework and mathematical framework to describe any technique that uses minimums or maximums.This allowed us to test different configurations in simulations.When we saw that this was very good and that it was also easily implementable in devices that exist in many laboratories, we started the experiments using one of the confocal microscopes that we had here”.And she concludes: “All the modifications that are needed, as well as the analysis software, are open source;we hope that many laboratories will start testing it to develop it further.”THE UNCOVERING FAIR ►#AltaData is over: this is your farewell (greetings from Indio Solari)The worst of the USA: they confess that they carry out coupsTour of claims in the patriotic weekendNelson Castro went crazy and attacked his Kirchnerist guestsViolence and delirium: the worst of the #9J marchCristina, Alberto and Massa: the meeting that anticipates what is to come.Vidal promises more adjustmentNew anti-Bullrich law on the Malvinas IslandsOwner: Talar Producciones SA CUIT: 33-71448833-9National Directorate of Copyright - IN PROCESS Edition Nº - 2815 - 07/21/2022Journalistic Director of El Destape Roberto Navarro - redaccion@eldestapeweb.comCommercial contact - comercial@eldestapeweb.com