Inverted Microscope: Introduction, Principle, Working and Applications

Inverted microscope introduction and its application

Introduction of Inverted Microscope

The inverted microscope was Invented in 1850 by a faculty member of the Medical College of Louisiana, named J. Lawrence. This is a reverse of the normal construction of a microscope, in which the objective lenses are found above the stage while the condenser and the light source are below the stage. Therefore the word, ‘inverted’was used. In this inverted microscope, we view the image from down, upwards whereas in a normal microscope viewing the image from up, downwards.

Principle of Inverted Microscope

The working principle of the inverted microscope is basically the same as that of a light microscope. In an inverted microscope, the source for transmitted light and the condenser are placed on the top of the stage as shown above picture, pointing down toward the stage. The objectives are located below the stage pointing up. The specimens or cells are observed through the bottom of the cell culture vessel. To meet the criteria for successfully inverted microscopy, the bottom of the culture vessel must have the highest optical features, which are given for the ibidi Polymer Coverslip and the ibidi Glass Coverslip.

Parts of the Inverted Microscope

However the parts of inverted the microscope are the same as those of other microscopes, the difference is the arrangement of these components, which are placed in inverted positions as shown above image.

It has the following components-

  • Stage: It has a large fixed stage able to hold large vessels e.g.  Petri plates.
  • Objective lens: It has movable objective lenses that are 4- 6 in the number of different magnification powers, which move on a vertical axis for viewing of the specimens.
  • Dual concentric knobs: They are fine and coarse adjustment knobs for fine-tuning and focusing the objectives to the specimen.
  • Nose piece: It is a rotating turret that holds the objectives.
  • condenser lens: It is used to concentrate light on the specimen.
  • Removable camera, fluorescent illuminators, scanners: They can also be temporarily attached

Working 

  1. Place the specimen in a slide or glass container on the stage.
  2. The stage does not move at all.
  3. Set the focus by moving the condenser and the objective lenses using the adjusting knob.
  4. The specimen can be observed through the ocular lens, or on the screen by attaching a video camera to it.
  5. The inverted microscope can be linked to, confocal scanners and fluorescent illuminators for different sets of experiments.

Application of Inverted Microscope

  1. It can be used to observe the living cells and tissues present at the bottom of any glass container such as a Petri plate, flask, etc. without preparing the slide, in its natural state.
  2. This inverted microscope is also used in certain diagnostic assays such as microscopic observation drug sensitivity (MODS) assay
  3. It is also useful in diagnostics fungal cultures e.g. detection of Phytophthora species in cultures.
  4. It is also helpful in parasitology for the diagnosis of nematology extraction specimens to observe nematodes such as Vermiform nematodes.

Bibliography

  1. https://www.wikilectures.eu/w/Inverse_microscope
  2. https://en.wikipedia.org/wiki/Inverted_microscope
  3. https://www.olympus-lifescience.com/en/microscopes/inverted/
  4. https://wiki.bugwood.org/Diagnostic_Uses_for_the_Inverted_Microscope
  5. https://www.quora.com/How-does-an-inverted-microscope-work
  6. https://ibidi.com/content/212-inverted-and-upright-microscopy#:~:text=Principle%3A,of%20the%20cell%20culture%20vessel.
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