Basics And Advantages Of Inverted Microscopy

In the scientific world, inverted microscopes are of great importance. It helps in the magnification of living cells through their culture vessels.  In 1850 the inverted microscope was invented by a faculty member of Tulane University, named J. Lawrence.  He constructed the inverted microscope from a normal and traditional trinocular compound microscope. The only difference was that all components placed in the inverted phase microscope were in an inverted fashion.  The pointing stage remains still or in a stationary position, while the condenser lens and objective lens are used for adjusting the focus of the images obtained. 

To put the construction of an inverted microscope,  it is the reverse construction of normal traditional microscopes. The objective lens is found above the pointing stage in the inverted microscopy technique, while the condenser lens and the light source are found below the stage.  This arrangement is in reverse order in the phase-contrast microscopy and traditional compound microscopes.  As far as the image formation is concerned, it is seen that the images formed in the inverted microscopes are better magnified than the other culture microscope techniques. Generally, an inverted phase microscope can give images of magnification of 10x to 40x.  All images are observed by the researchers and scientists in a downward fashion.  This means,  all cells are observed first from the bottom and then from the upper region.

With phase-contrast microscopy also, the inverted microscope components can be combined to make the inverted phase-contrast microscope. These microscopes are all good at giving out the best, and high contrast images of all biological samples. In this process, it recruits the process of reflection and diffraction from the phase objects to generate images.  Though the interferences caused by the light rays are not that subtle to be noticed from the naked eyes. Still, it makes the right image formation. 

Basic Component Of Inverted Microscope

Like a trinocular compound microscope,  the inverted microscope also has the same components, but in reverse order.  It also shares its components with the composite microscopes, all in reverse order. As the name suggests,  the inverted microscope has all basic components in an inverted fashion than the upright and inverted microscope. Because of its inverted position of all basic components, it is called an inverted microscope. When it is used for examining or observing cells of tissue culture, they are called cell culture microscope, tissue culture microscope, and culture microscope. 

Following are the basic components of an inverted microscope that are used for  generating magnified images :

1. Pointing Stage

This is the landing part of the inverted microscope. All biological samples and specimens are placed over them for magnification and observations. In culture microscopes, the culture vessels are placed on the stage areas for observations.

2. Pointing Stage Clips

The pointing stage clips are used for holding the biological specimens on the stage for magnification. Sticking to the stage in a proper position is important for the magnification process. These clips hold the specimens tight to the right position.  

3. Microscope’s Arm

This part of the trinocular inverted microscope is important for holding the mechanical and optical parts. Like light source and condenser and objective lens. These are supported by the inverted microscope’s arm.

4. Objective Lens of Inverted Microscope

The objective lens of the inverted microscope produces and magnifies the images of the biological specimen. It moves along the vertical axis of the microscope to produce the images.

5. Dual Concentric Knobs

The inverted microscope contains fine and coarse adjustment knobs for obtaining good images. It helps in fine-tuning and focusing the objectives of the specimen. With the help of concentric knobs, the focus can be adjusted by the researcher and scientist in a single go. By this adjustment, a fine image can be obtained through the inverted microscope.

6. Nosepiece

Nosepiece holds the objective lens of an inverted microscope.  Without the use of a nosepiece, the objective lens cannot be held firmly.

7. Condenser lens

A condenser lens is known for mediating the light rays to the biological specimens.  The condenser lens concentrates the light on the specimen for better observations.

8. Digital Camera

Many advanced upright and inverted microscopes come with a digital camera. The use of the digital camera is to record or capture the image of the specimens or samples.  In the market, a camera for a trinocular microscope comes at reasonable prices. The best trinocular microscope has a digital camera for recording the specimens.

Advantages Of Inverted Microscopy Techniques

Following are the advantages of inverted microscopy techniques:

• The Living cells can be observed by using an inverted microscope with a camera.
• Kohler Illumination can be used with Inverted Microscopy techniques for better magnification.
• Differential Interference Contrast of phase contrast microscopy can be used with an Inverted Microscope. This gives out the high contrasted images of biological samples.
• Phase-contrast optics can be used with the principle of an inverted microscope, to increase the image quality.
• Large and heavy biological specimens like metallurgical samples can be observed in the Inverted Microscopes. They are capable of observing the samples in minute detail.
• The microscopy technique allows researchers and scientists to attach a variety of digital recording equipment. Like a digital camera, it records the specimen‘s images with details.
• Inverted microscope is famous for its maintenance of sterility of all biological samples. It maintains the sterility of all biological samples, it can be in its pure state after observations also.
• The inverted microscope is useful in the process of parasitology diagnosis. 

Some Types Of Inverted Microscopes

There are some different types of inverted microscopes that are used in the scientific world.  They are as follows :

1. Biological Inverted Microscopes

The biological inverted phase microscopes are used for observing  living cells and organisms at the bottom of the culture vessel or culture flask. It is seen that the live cell analysis requires larger containers filled with liquid. This provides natural conditions for all types of biological specimens.  These samples are viewed under the upright and inverted microscope with a camera. The fluorescence sources can also be used for the observations of live cells.

2. Metallurgical Inverted Microscopes

The inverted microscopes are not only limited to biological observations, but they work in metallurgical areas as well.  With the help of a metallurgical microscope,  one can view the metal and solid particles in the metallurgical factory.   It also helps in the micromanipulation process.

Limitations Of Inverted Microscope

It is no doubt that inverted microscopes are good for magnifying the images of biological samples.  But, it also comes with certain limitations that should not be neglected.  Following are the list of limitations:

1. Generally, the inverted microscopes are very expensive to acquire for the lab. You can get the trinocular microscope price from the offline and online mediums as well.
2. Inverted microscopes are manufactured by very few companies because they are expensive to manufacture. So, there is a shortage of these microscopes than the other forms of microscopes.
3. Inverted phase contrast microscopes are rarely found in the market for purchase and usage. So, the researchers and scientists may find trouble in finding one.
4. In an inverted microscope, it is difficult viewing the specimens through thick glass vessels such as Petri plate dishes.  Hence, they require very high optical quality and a good digital camera.

Conclusion

The inverted microscopy techniques are useful in magnifying images of biological samples.  The instrument has basic components same as the normal microscopes but in reverse order.  Due to its good feature of magnification, it has multiple applications in the biological world.