Inverted microscope Definition
Inverted microscopes are almost similar to a simple microscope, but all the components are placed in the inverted condition.
The Inverted microscope comes with its own light source and a condenser lens. Which are located at the top portion of the microscope and is pointing down. Whereas the objectives and the turret are located below the stage, pointing upwards.
In this microscope, we observe the specimen from down, (upwards) instead of from the up.
Inverted microscopes come with three to six objective lenses. The magnification power of these lenses are ranging from 4x to 40x.
In 1850, a faculty member of Tulane University J. Lawrence Smith, first invented the Inverted microscope.
Principle of Inverted Microscope
An inverted microscope follows the same working principle, of a simple microscope or upright light microscope. But in an inverted microscope the light source and condenser lens located at the top of the stage and pointing down to the stage.
The condenser lens at the top of the stage helps to focus the light rays on the specimen. Where the objective lens located are located below the stage pointing up. In this microscope, the cell cultures are observed through the bottom of the cell culture vessel.
Parts of Inverted Microscope
All components of an inverted microscope are similar to a composite microscope. But the main difference is, all the components remain in the inverted condition. An inverted microscope contain these following components;
- Stage: Specimen placed over it.
- Stage Clips: It holds the specimen in position.
- Arm: It holds the optical and mechanical parts of the microscope.
- Objective Lens: It produces and magnify the images of the specimen. It moves along the vertical axis.
- The Dual concentric knobs: It contains fine and coarse adjustment knobs which helps in fine-tuning and focusing the objectives to the specimen.
- Nosepiece: It holds the objective lens.
- Condenser lens: It concentrates the light on the specimen.
- Digital Camera: It used to record or capture the image of the specimen.
Operating Procedure of Inverted Microscope
- Placed the inverted microscope on a table in stable condition.
- Then placed the specimen containing slide or glass container on the specimen stage.
- The specimen stage does not move at all.
- Now adjust the focus by moving the objective and condenser lens (use the knobs to control the movement of Condenser and Objective Lens).
- The image of the specimen can be observed by the ocular lens or on a digital screen.
- The inverted Microscopes can be used with confocal scanners and fluorescent illuminators.
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The capability of Inverted Microscopes
Inverted Microscopes are useful for metallurgical samples, and for the observation of living specimens or tissues.
Cell division process can be observed by using this microscope which is not possible by using a conventional compound microscope. Living cells can be observed for a long time by using this microscope, which makes it more useful.
The viewing of valuable life processes can be researched longer. This is its major advantage over a compound light microscope.
Large specimens required to keep in a large petri dish for viewing under inverted microscopes instead of on a slide. The container should be covered for less evaporation and more gas exchange to maintain life.
Application of Inverted Microscopes
- Used for the detection of Phytophthora spp in culture.
- It is used to observe the living cells and organisms at the bottom of a large container (e.g., a tissue culture flask).
- It is also used to examine the mycobacterium tuberculosis.
- This microscope also used in micromanipulation.
Advantages of Inverted Microscopy
- Living cells can be observed by using inverted microscopes.
- Kohler Illumination can be used with Inverted Microscopes.
- Differential Interference Contrast can be used with an Inverted Microscopes.
- Phase-contrast optics can be used with it, to increase the image quality.
- Large and heavy specimens like metallurgical samples can be observed in the Inverted Microscopes.
- It allows us to attach a variety of digital recording equipment.
- It maintains the sterility of the specimens.
Disadvantages of Inverted Microscopy
- It is very expensive.
- Lack of Manufacturing companies.
- Specimens can not be observed through the thick glass.