BioFilm
Characterization of microorganisms, such as bacteria, fungi, and algae, can be challenging for a variety of reasons. They move constantly, their life cycles are relatively short, and they can be adversely affected by common laboratory handling and measurement preparation protocols. Advances in microscopy have made the measurement of these organisms easier and reduced some of the barriers to successful measurements, but roadblocks persist.
Biofilms are everywhere around us. They are composed of colonies of microorganisms that literally “stick together” and then adhere to a large and diverse number of surfaces—sometimes having a beneficial effect, but most often not. In medical research and development, biofilm presence and growth contribute to chronic wounds that will not heal to a costly spectrum of dental issues, prosthetic implant infection and much more.
And it is not just life science that meets the remediation and/or elimination of these stubborn colonies head on. Environmental and wastewater treatment, industrial bioprocessing, the food industry, pharmaceuticals, agriculture and research need to understand how the biofilms grow as well as how they develop the ability to resist antibiotics, chemicals, and other stressors.
Measurement Options for Biofilms
Other optical profilometry techniques have been used extensively in biofilm characterization. This technology is effective in managing fluorescence to produce sharp, well-focused, 3D images of scattering media that may be particularly thick. It is, however, a slow process and requires multiple images to be stitched together to get a clear visualization of the entire biofilm colony.
Scanning Electron Microscopes (SEM) have also been used to measure biofilms, but its main drawback is it cannot be used on living samples. Atomic Force Microscopes (AFM) can also be used, but the instruments tend to be outside of laboratory budgets and require extensive operator training.
3D Optical Profilers with SWLI
More recently, scanning white light interferometry has shown great promise in measuring biofilms in real time as detailed in the research 'Vertical growth dynamics of biofilms' (PNAS, 2022), looking at surface topology, growth factors, media/colony interfaces, and optimal media compositions. It was also possible to observe a time-lapse of bacteria during the initial growth and division phases. These measurements provided insight into bacterial behavior and growth processes.
All of this was done with the Zygo ZeGageTM Pro 3D optical profiler, which is a high resolution, benchtop powerhouse. It measured the biofilm quickly, in some cases in less than 5 seconds, is non-contact, and does not require sample preparation. Unlike other optical profilometry techniques, the ZeGage was able to measure the entire sample at the same time. Mx software offers a comprehensive suite of graphics tools to visualize data, presenting results as detailed 2D and 3D surface maps and profiles.
Research labs in universities, government, biotechnology, food, agriculture, and industrial bioprocessing now have an alternative to characterizing biofilms.
Case Study: Leading University Research Lab uses ZygoTM Optical Profilers for Faster, More Detailed Biofilm Insights
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