Up to now, if scientists wanted to study blood cells, algae, or bacteria under the microscope, they had to mount these cells on a substrate such as a glass slide. Physicists at Bielefeld and Frankfurt Universities have developed a method that traps biological cells with a laser beam enabling them to study them at very high resolutions. In science fiction books and films, the principle is known as the ‘tractor beam’. Using this procedure, the physicists have obtained superresolution images of the DNA in single bacteria.
One of the problems facing researchers who want to examine biological cells microscopically is that any preparatory treatment will change the cells. Many bacteria prefer to be able to swim freely in solution. Blood cells are similar: They are continuously in rapid flow, and do not remain on surfaces. Indeed, if they adhere to a surface, this changes their structure and they die.
‘Our new method enables us to take cells that cannot be anchored on surfaces and then use an optical trap to study them at a very high resolution. The cells are held in place by a kind of optical tractor beam. The principle underlying this laser beam is similar to the concept to be found in the television series “Star Trek”,’ says Professor Dr. Thomas Huser. He is the head of the Biomolecular Photonics Research Group in the Faculty of Physics. ‘What’s special is that the samples are not only immobilized without a substrate but can also be turned and rotated. The laser beam functions as an extended hand for making microscopically small adjustments.’
The Bielefeld physicists have further developed the procedure for use in superresolution fluorescence microscopy. This is considered to be a key technology in biology and biomedicine because it delivers the first way to study biological processes in living cells at a high scale – something that was previously only possible with electron microscopy. To obtain images with such microscopes, researchers add fluorescent probes to the cells they wish to study, and these will then light up when a laser beam is directed towards them. A sensor can then be used to record this fluorescent radiation so that researchers can even gain three-dimensional images of the cells.
In their new method, the Bielefeld researchers use a second laser beam as an optical trap so that the cells float under the microscope and can be moved at will. ‘The laser beam is very intensive but invisible to the naked eye because it uses infrared light,’ says Robin Diekmann, a member of the Biomolecular Photonics Research Group. ‘When this laser beam is directed towards a cell, forces develop within the cell that hold it within the focus of the beam,’ says Diekmann. Using their new method, the Bielefeld physicists have succeeded in holding and rotating bacterial cells in such a way that they can obtain images of the cells from several sides. Thanks to the rotation, the researchers can study the three-dimensional structure of the DNA at a resolution of circa 0.0001 millimetres.
Professor Huser and his team want to further modify the method so that it will enable them to observe the interplay between living cells. They would then be able to study, for example, how germs penetrate cells.
Learn more: Optical tractor beam traps bacteria
A naturally occurring predatory bacterium can work with the immune system to clear multi-drug resistant Shigella infections in zebrafish.
It is the first time the predatory bacterium Bdellovibrio bacteriovorus has been successfully used as an injected anti-bacterial therapy and represents an important step in the fight against drug-resistant infections, or ‘superbugs’.
Shigella infection is responsible for over 160 million illnesses and over 1 million deaths every year – and is a common cause of ‘travellers’ diarrhoea.’ Cases of drug-resistant Shigella are also on the rise as, although the diarrhoea usually clears up without treatment, antibiotics are often used even in mild cases to stop the diarrhoea faster. Resistance to antibiotics has prompted a team of researchers from Imperial College London and Nottingham University to look to the natural environment for creative solutions to this problem.
‘PREDATORY ACTION’ – ABOUT THE STUDY
To investigate Bdellovibrio’s ability to control drug resistant Gram-negative infections, researchers injected zebrafish larvae with a lethal dose of Shigella flexneri strain M90T which is resistant to both streptomycin and carbenicillin antibiotics. Bdellovibrio was then injected into the larvae’s infection site, and a decrease in the number of Shigella was seen. In the absence of Bdellovibrio, zebrafish were unable to control the replication of Shigella and levels of the bacteria rose.
Co-lead author Dr Serge Mostowy from the Department of Medicine at from Imperial College London said: “This study really shows what a unique and interesting bacterium Bdellovibrio is as it presents this amazing natural synergy with the immune system and persists just long enough to kill prey bacteria before being naturally cleared. It’s an important milestone in research into the use of a living antibiotic that could be used in animals and humans.”
Bdellovibrio can invade and kill a range of Gram-negative bacteria, such as E. coli and Salmonella, in the natural environment. Previous research has shown that it can reduce pathogen numbers in the stomach of chickens when taken as an oral therapy, but there is growing need to develop therapies to target infections in wounds and organs. Successful use of Bdellovibrio highlights its potential uses in tackling a range of drug-resistant Gram-negative bacterial infections that can develop in hospital patients.
The urgent requirement for new antimicrobials calls for increasingly creative solutions
– Dr Alex Willis
Co-lead author from Imperial’s Department of Medicine
Dr Mostowy and co-lead author Dr Alex Willis from the Department of Medicine at Imperial said: “The urgent requirement for new antimicrobials calls for increasingly creative solutions. The zebrafish has been a fantastic model for us to generate rapid understanding of how a living antibiotic can work in an animal. Our findings here provide the basis for testing Bdellovibrio in higher vertebrates and ultimately, humans.
“The translucent zebrafish facilitates powerful microscopy that enables striking understanding more difficult to achieve using other animal models. Being able to visualise the infection as well as the predatory bacteria and host immune cells has been invaluable in helping us understand how Bdellovibrio can function in an animal.”
Professor Liz Sockett, co-lead author from The University of Nottingham said: “This has been a truly ground-breaking collaboration that shows therapeutic Bdellovibrio in action inside the translucent living zebrafish. The predatory action of the Bdellovibrio breaks the Shigella-pathogen cells and this stimulates the white blood cells; redoubling their ‘efforts’ against the pathogen and leading to increased survival of the zebrafish ‘patients’.”
Remarkably, Bdellovibrio is also able to reduce pathogen load in immunocompromised zebrafish larvae that have been depleted of white blood cells. However, survival is significantly greater in immune-competent zebrafish, showing that Bdellovibrio’s maximum therapeutic benefit comes from its ability to work cooperatively with the host’s own immune system.
Dr Michael Chew, Science Portfolio Advisor at Wellcome said: “It may be unusual to use a bacterium to get rid of another, but in the light of the looming threat from drug resistant infections the potential of beneficial bacteria-animal interactions should not be overlooked. We are increasingly relying on last line antibiotics, and this innovative study demonstrates how predatory bacteria could be an important additional tool to drugs in the fight against resistance.”
Severe gum disease known as periodontitis can lead to tooth loss, and treating it remains a challenge. But new approaches involving silicon nitride, a ceramic material used in spinal implants, could be on the way.
The surface of silicon nitride has a lethal effect on the bacteria that commonly cause periodontitis. Now scientists, reporting in ACS’ journal Langmuir, are examining why this happens. Their findings could help inform future efforts to treat the disease.
About half of American adults have some form of gum disease. It’s caused by bacteria that infect the tissue around teeth, resulting in gum inflammation. If the condition progresses, the bacteria can damage the bone that supports the teeth. In addition to tooth loss, periodontitis can increase a person’s risk of heart attack or stroke. Options for treatment include scaling and root planing, topical antibiotics and surgery. Giuseppe Pezzotti and colleagues wanted to find a new alternative by studying the reactions of bacteria to antimicrobial silicon nitride.
Individual bacterial cells have short memories. But groups of bacteria can develop a collective memory that can increase their tolerance to stress. This has been demonstrated experimentally for the first time in a study by Eawag and ETH Zurich scientists published in PNAS.
Bacteria exposed to a moderate concentration of salt survive subsequent exposure to a higher concentration better than if there is no warning event. In populations exposed to a warning event, survival rates upon a second exposure two hours after the warning are higher than in populations not previously exposed. Firstly, salt stress causes a delay in cell division, leading to synchronization of cell cycles; secondly, survival probability depends on the individual bacterial cell’s position in the cell cycle at the time of the second exposure.