infections and billions of dollars in excess health care costs are related to healthcare-associated infections across the U.S. every year¹

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Finally, dark field microscopy requires no particular set-up or expensive equipment and a light microscope can be converted into a dark field microscope with minimal additional costs.

This standard and elements of performance address the necessary structures and activities for implementing an effective antibiotic stewardship program. Requirements focus on appointing a program leader, convening a multidisciplinary oversight committee, providing staff education and training, monitoring antibiotic use and resistance patterns, and implementing evidence-based strategies to optimize prescribing practices and improve antibiotic stewardship activities.

is the estimated annual national cost to treat infections caused by six antimicrobial-resistant germs frequently found in health care¹

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Dark field microscopy differs from conventional microscopy, producing different styles of images that lend themselves to different applications. Not all applications of conventional microscopy apply to dark field microscopy, and not all applications of dark field microscopy work as well with conventional techniques.

This standard and elements of performance focus on the hospital’s governing body being accountable for the infection prevention and control program, ensuring its implementation, performance, and sustainability.

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The images usually appear as bright white against a dark background, regardless of the color of the sample. Pigments in specimens are viewed as false colors, in that the color they are depicted as in the image does not accurately reflect the actual color of the object.

Joint Commission accreditation standards provide a framework that challenges healthcare organizations to strengthen their infection prevention and control practices. They guide healthcare professionals on the most effective approaches to patient care, infection prevention, and safety.

C Robert Bagnall. (2012) Dark Field Microscopy [Online]. UCI Department of Chemistry. Available at: https://www.chem.uci.edu/~dmitryf/manuals/Fundamentals/Dark%20Field%20microscopy.pdf (Last accessed October 2022).

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Next, the samples must be exposed to intense illumination, which can cause damage to the sample depending on what is being imaged.

Moore, Sarah. "The Benefits and Limitations of Dark Field Microscopy". AZoOptics. https://www.azooptics.com/Article.aspx?ArticleID=2335. (accessed December 19, 2024).

As a result, dark field microscopes have become an established tool in microbiology laboratories for use in various applications, including the visualization of clinical samples of spirochetes (e.g., Treponema palladium [syphilis], Borrelia burgdorferi [lyme borreliosis], and Leptospira interrogans [leptospirosis]), the observation of microbial motility (e.g., visualizing the tufts of bacterial flagella), and viewing the internal structure of eukaryotic microorganisms (e.g., algae and yeast).

Moore, Sarah. "The Benefits and Limitations of Dark Field Microscopy". AZoOptics. 19 December 2024. .

Dark field microscopy is a technique invented in 1830 for the observation of living, unstained cells and microorganisms. This type of microscopy requires intense illumination of the sample within a dark surrounding, making it appropriate for studying particular samples. While its limitations have seen it fall behind more modern microscopy methods, in recent years, it has gained popularity as scientists have begun to use it alongside other methods.

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Oliviero, F., Punzi, L. (2022) Basics of Polarized Light Microscopy. In: Mandell, B.F. (eds) Synovial Fluid Analysis and The Evaluation of Patients With Arthritis. Springer, Cham. https://doi.org/10.1007/978-3-030-99612-3_9

Finally, specimens must be thin. Dense samples can negatively impact the contrast and accuracy of images produced. For this reason, not all samples are appropriate for dark field imaging.

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Dark field microscopy can often image samples unsuitable for conventional bright microscopy. Compared with bright field microscopy, dark field obtains greater resolutions, thus imaging samples in greater detail.

Healthcare-associated infections (HAIs) significantly threaten patient safety, causing longer hospital stays, higher medical costs, and increased mortality. Effective prevention, including strict hand hygiene, proper equipment disinfection and sterilization, and PPE use, is essential. Effective stewardship programs help preserve the efficacy of existing antibiotics, improve patient outcomes, and decrease the spread of resistant bacteria. Ensuring safety requires collaboration between healthcare workers and patients to follow these measures.

The intensity of light required for dark field microscopy can also lead to glare and distortion. These issues also make the technique unreliable in obtaining specimen measurements.

Sekine, R., Moore, K., Matzke, M., Vallotton, P., Jiang, H., Hughes, G., Kirby, J., Donner, E., Grovenor, C., Svendsen, C. and Lombi, E. (2017) Complementary Imaging of Silver Nanoparticle Interactions with Green Algae: Dark-Field Microscopy, Electron Microscopy, and Nanoscale Secondary Ion Mass Spectrometry. ACS Nano, 11(11), pp.10894-10902. https://pubs.acs.org/doi/full/10.1021/acsnano.7b04556

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This standard and elements of performance focus on hospital-wide infection prevention and control program for the surveillance, prevention, and control of healthcare-associated infections (HAIs) and other infectious diseases. The elements of performance address the program’s leader(s) and responsibilities, and requirements for the program’s policies and procedures and the scope.

Dark field microscopy also generates an improved image contrast vs bright field microscopy without the need to stain samples, thus, not killing cells.

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While there are numerous essential benefits of dark field microscopy, it is not without its limitations. When visualizing wet specimens of live organisms, it is essential to do this rapidly to ensure that movement does not interfere with the quality of the image.

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As a result, viewing aquatic organisms with refractive indices similar to the surrounding water has become a standard sample studied with dark field microscopy. Other biological samples make ideal candidates for this method, such as bacteria, tissue cultures, yeast, and others. Non-biological samples suitable for study with dark force microscopy include chemical and mineral crystals and thin segments of polymers.

Another significant benefit of dark field microscopy is that no sample preparation is required (e.g., no staining), which reduces the time demands of the process.

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Moore, Sarah. 2022. The Benefits and Limitations of Dark Field Microscopy. AZoOptics, viewed 19 December 2024, https://www.azooptics.com/Article.aspx?ArticleID=2335.

Further limitations of dark field microscopy include its sensitivity to contaminants. Specimen slides must be meticulously cleaned to ensure that dust and dirt are eliminated from the area and such contamination can distort the images produced.

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This standard and elements of performance focus on the hospital’s implementation of its infection prevention and control program, including a proactive risk assessment to help organizations determine the scope of surveillance and implement the appropriate level of infection prevention and control activities.

This standard and elements of performance focus on protocols to enhance hospitals’ and critical access hospitals’ preparedness for high-consequence infectious diseases or special pathogens.

In dark field microscopy, an opaque disk is placed beneath the condenser lens, ensuring that only light scattered from the specimen is allowed to reach the eye.

A colony of green planktonic algae Volvox under the dark field microscope. Image Credit: D. Kucharski K. Kucharska/Shutterstock.com

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Dark field microscopy is most useful when researchers need to visualize unstained, transparent specimens. As a general rule, specimens appropriate for viewing with dark field microscopy should have a reflective index similar to the surroundings. Such a refractive index makes a sample unsuitable for viewing with conventional bright field microscopy.

While dark field microscopy has been around for over a century, its limitations have hindered its ability to keep up with the vast advances in alternative microscopy techniques, such as phase contrast and differential interference contrast (DIC) microscopy. For this reason, the technique is not commonly used in modern imaging. However, it has been increasingly combined with modern methods such as fluorescence microscopy in recent years.

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Instead of passing through the specimen, as with other forms of microscopy, in dark field, the light is reflected by particles on the slide. Light is directed so that it passes through the condenser’s outer edge at a wide angle and hits the specimen at an oblique angle.

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Moore, Sarah. (2022, October 19). The Benefits and Limitations of Dark Field Microscopy. AZoOptics. Retrieved on December 19, 2024 from https://www.azooptics.com/Article.aspx?ArticleID=2335.