Microscopy is a technical field that allows us to view and examine objects that can’t be seen by the naked eye.
Using a microscope to do so, this aids scientists massively when it comes to scientific breakthroughs, identifying diseases, and when observing biological processes and cell formation and structure.
However, for researchers and scientists to be successful in this technical field, they have to use the right techniques.
A variety of microscopy imaging techniques are utilized by researchers and scientists to enhance their ability to view the microscopic world.
If you’re a keen scientist or researcher, you may already know a lot about the techniques employed. If not, you’ve come to the right place.
To improve your understanding of the microscopy imaging techniques used, we’ve outlined all the ones you need to know about.
We’ll take you through each one and explain the benefits and disadvantages.
Let’s get started!
Brightfield Microscopy
The most elementary microscopy imaging technique is the brightfield Microscopy technique.
Easily allowing researchers and scientists to view various specimens, this technique is normally used when working with a compound microscope.
The technique takes dark specimens and contrasts them by applying a bright viewing field.
The light that surrounds the specimen is very beneficial as it makes the user’s view of each specimen much clearer.
This technique is so commonly used with compound microscopes that the compound microscope is often referred to simply as the brightfield microscope.
The only disadvantage to this technique is that it can introduce irrelevant details that shouldn’t be present in the specimen.
Oil Immersion Microscopy
Next up, we have the oil immersion microscopy technique. This technique is essential when it comes to examining and observing specimens with compound microscopes.
As a result of significant disadvantages, great care must be taken when using the technique in order to minimize any issues. The main disadvantage to avoid is cement drying on the lens.
A suitable mix of synthetic oils helps create the perfect viscosity. This viscosity then helps researchers achieve high-resolution, high-quality images. Bacteria, striated tissue, and inanimate objects can then be easily studied.
Similar refractive indexes also offer large bright images.
Darkfield Microscope
Researchers typically use the darkfield microscope technique when trying to make objects appear brighter against a darker background. In fact, this technique is the best to use in this scenario.
If the researcher wasn’t to use this technique in these circumstances, the refractive values would be too similar, making it difficult to view each specimen.
Therefore, this technique’s biggest advantage is being able to see the specimen clearly.
The main disadvantage of this technique is its sensitivity to contaminants and the need for intense amounts of light.
Darkfield microscopy is achieved by making modifications to the microscope.
Kohler Illumination
The next technique on our list was originally developed in 1893. It was through optimization of the microscope’s optical train that homogeneously bright light was created without creating glare and artifacts.
This technique’s main advantage is a uniformly illuminated specimen and high levels of contrast. This again assists researchers and scientists hugely when it comes to clearly examining specimens.
The main downside to this technique is its difficulty to achieve an evenly illuminated specimen at low power.
Differential Interference Contrast (DIC) Microscopy
The differential interference contrast microscopy technique transforms the smallest of differences in refraction indexes of the light that passes through your unstained specimens.
It also transforms the optical path differences in regard to the specimen’s shape. It will transform the specimen’s shape into a monochromatic shadow-cast, enabling clearer, easier observation.
While it can be used to observe living specimens, the technique better suits thicker objects. One of the main uses of the DIC technique is to observe the crystal growth process.
It is worth noting that this technique is quite expensive as a large number of prisms are needed.
Phase Contrast Microscopy
The next microscopy technique is phase contrast microscopy. This technique involves using diffraction and interference to add light and dark contrast to transparent specimens.
One of the best things about the technique is that no staining is required. Therefore, live specimens can be viewed.
The phase contrast microscopy technique is primarily used to image blood, living cells, and cultured cells.
Although the technique provides well-contrasted, highly detailed images, it isn’t uncommon for halo distortions and shade-off to occur.
The ability to observe live specimens in their natural state outweighs these disadvantages though.
Fluorescence Microscopy
This technique uses high-powered light waves that offer researchers unique viewing points. The viewing points on offer aren’t available when using conventional light microscopes.
To provide an even better view, this microscopy technique uses stains. This allows researchers to view more details at once, including the inner structure of most cells.
Compared to other techniques, the fluorescence microscopy technique provides 3D images of specimens and superior image quality.
The main downside to the technique is that prolonged exposure to the fluorescent light can result in bleaching. A loss of intensity can also occur.
Confocal Microscope
Confocal microscope techniques require state-of-the-art technology in order to work. The technique involves separating the light waves with lasers.
By doing so, scientists and researchers are left with images that offer higher resolution and no blurry edges.
Using this technique also allows researchers to take more images in a shorter space of time, allowing for smaller sections of the specimen to be viewed at one time.
Of course, the only downside to this microscopy technique is how much it costs. The expense of the state-of-the-art equipment needed to perform the technique isn’t cheap.
Polarizing Microscope
Finally, we have the polarizing microscope technique. This technique is the best option if you’re working with birefringent materials.
The technique uses polarization to enhance the overall color and contrast of images, providing researchers with detailed information regarding the absorption, composition, and structure of specimens.
This technique mostly benefits the study of minerals and rocks in the petrography and geology fields. Biology and medicine also benefit from this imaging technique.
The main issue with this technique is that it only uses a small amount of each specimen. Therefore, false results or sample bias can occur if the researcher isn’t careful.
Final Thoughts
That concludes this post, looking at the microscopy imaging techniques you need to know about.
Microscopy imaging techniques make it much easier and possible for researchers and scientists to learn more about various things.
Whether it be examining a new disease, or the studying of crystals, using the right microscopy techniques can make specimens clearer to see, easier to take images of, and much sharper.
Now that you’ve made your way through each technique, you should have a much better understanding of how microscopy techniques work and how they can be used to enhance a range of microscopy experiments.
If you practice microscopy, why don’t you take some of these techniques and see how you can put them to good use!
Ever since I was a little girl, I have been fascinated by science. Every holiday I would ask for the newest science kit, and spend weeks learning with it.
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