Medical and biological sciences research are now focused on the real time, real life observation of living organisms to analyze its components and understand life better than we already do. Specifically designed for live specimens, this type of microscope enables observation of virtually invisible occurrences within a cell.
Obviously more advanced than the typical light microscope, this instrument uses the various refractive abilities of different objects to heighten the contrast between colorless and transparent structures. Under this powerful type of microscopy, specimens remain alive as they do not need to be stained - colored or dyed - to distinguish characteristics.
Light Waves and Visible Contrasts
Light waves peak and valley in regular intervals. The moment they line up, they're said to be in phase; otherwise they're said to be out of phase. This basic understanding of light waves is the key to how a phase contrast microscope works – it influences the optical path of light through transparent or colorless objects under observation.
Light waves passing through transparent portions of the specimen is slower then the uninfluenced light. To cause a difference in brightness, a transparent phase-plate mounted on the microscope increases the phase to half a wavelength, making the transparent object outshine its surroundings. This unique feature of phase contrast microscope makes it an indispensable tool in medical and biological observations of living cells.
How It Works
A phase contrast microscope makes use of two light sources – one under your specimen and another that is reflected off your specimen. Now, light passes through any transparent object but gets reflected off the surface of a solid, colorless object. When these light waves condense on a lens placed just above your specimen, it is easy to see if the light waves are in or out of their phase. This phase condenser lens on this type of microscope is analogous to the fluorescent filters in fluorescence microscopes; both make light differentiation possible.
Advances in Biomedical Microscopy Technologies
The latest in biomedical microscopy is the introduction of infinity-corrected phase contrast microscopes. Over the last decades, the largest manufacturers of research and/or professional grade microscopes have introduced this new type of optical system into various microscopy modules. This technology places another lens between the objective and the eyepiece and sets the object image to infinity. This correction makes it possible to introduce auxiliary components in the microscope.
This integrated capability makes imaging a an exciting part of the research. Now, you can capture, store, display – virtually do anything – with images of your specimen. With the flexibility of integrated digital technology, you can capture full color photographs or videos of your specimen and display, store and retrieve them from your computer. These capabilities produce virtually the same images on eyepieces and photo ports, allow you to hook up various types of cameras on to the microscope, which would definitely make training and demonstration more real life.
Inverted Phase Contrast Microscope
This article, however, will only focus on the lenses system - the whole group responsible in forming the image.
The Eyepiece
Without this part, you cannot see the specimen being examined. This is by far, among the most important parts of the microscope. This is a cylindrical tube which contains a lens. Stereo microscopes have two eyepieces, while a compound microscope only has one.
The Objective Lenses
The primary role of the objective lenses is to gather light from the object or specimen being observed. There are several objective lenses in a microscope. Each lens varies according to magnification to provide you with more options for better specimen examination.
Sometimes, special objectives are used for certain examination techniques. The phase contrast - developed by Frits Zernike - is by far the most sophisticated. The phase contrast technique is basically about enhancing contrast. As a general rule, you can easily differentiate two specimens or even objects if they have high contrast - white is better seen when set against a black background.
Books and even Internet articles that explain the phase contrast method seems too technical for this kind of article - one that's focused in explaining the lenses system of the microscope. Still, to give you an idea on the concept of the phase contrast microscopy, this simple explanation would do:
It's a bit hard to have a microscopic view of living tissues (such as blood) because most of them are transparent. These kinds of specimen are usually called phase objects because they do not absorb light - instead, they diffract (to cause the bending of the waves) them. As a result of the bending, the light becomes less visible (by 1/4 wavelength). Let's not discuss what 1/4 wavelength really means but simply put, it suggests that such a reduced property of light cannot be seen by your naked eye, not even by a camera.
It is in this context that the phase contrast technique becomes very useful. Somehow, this technique - using special objectives and a condenser - is able to solve such a problem. It is able to yield an image that is darker than its light background.
Other special objectives are the infinity-corrected optical systems - where the image distance is set at infinity. These lenses are great for biomedical and industrial microscopes.
The Nosepiece
As mentioned, most microscopes have more than one objective lenses. As such, these objectives need something to hold them together - and this is where the nosepiece comes in handy.
The Tube
This is the very thing that holds the eyepiece and the objective lenses - but at an appropriate distance. This is responsible for keeping away stray light.
While the lenses are already powerful enough, some still prefer to use accessories such as a fluorescent filter. This is done in order to get optimum lighting and for special viewing techniques.
Once again, these parts only refer to the lenses system of a microscope. These are the ones responsible in forming the image of the specimen. You still need to acquaint yourself with the other parts - such as the illumination system and specimen control to better understand how the microscope works.
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