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Understanding Phase Contrast Microscopes

In 1953, the world recognized the scientific genius that is the phase contrast microscope. Forget high useful - it wasn't just useful, it was crucial for actual live observations of intra-cellular processes like cell division.

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.

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An objective lens of a light microscope is marked as 45. What is the total magnification when the objective le?

Question 2. Describe the functions of following parts of microscope: Objective lens, Coarse and Fine adjustment Knobs, Illuminator, Iris diaphragm, and Condenser.
Question 3. What changes do you expect to observe (image-size, light, field of view, depth, resolution, etc.) if you change the magnification of a microscope from lower to higher?
Question 4. Compare the advantages and disadvantages of observing a specimen under the scanning lens of 40X and 400X magnification.
Question 5. How does dialysis differs from osmosis?

1. Assuming you are using a 10X eyepiece, the total magnification is the power of the eyepiece times the power of the objective lens or 450X.

2. Objectives: magnify and form the image of the object being observed
Coarse and fine focus knobs: raises or lowers the stage to bring the object being observed into focus. The coarse is for initial focus. The fine is to observe structures by adjusting the height of the stage.
Illuminator: the source of light to examine the structures of the specimen
Iris diaphragm: controls the amount of light striking the object being observed. It increases or decreases the contrast of the specimen.
Condenser: focuses the light from the illuminator to a fine point to illuminate the object being observed.

3. As you increase the magnification, the image of the object gets larger, the field of view, depth of focus decreases, the resolution increases, the light intensity decreases.

4. Lower magnification allows one to see large areas of the specimen in a single field of view. The higher magnification allows one to observe fine or small details of the specimen as well as position in relationship to other structures within the specimen.

5. In dialysis of a substance, the membrane is permeable to specific molecules. In osmosis, only water is permeable.

Olympus Launches BX3 Microscope Series
Olympus has unveiled its BX3 line of upright clinical and research microscopes with exclusive ergonomic and imaging features that provide unprecedented comfort, ease of use and accuracy. The all-new BX3 microscope lineup will deliver outstanding optical performance, value, reliability and eco-friendly operation.

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