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Inverted Microscope – Principle, Parts, Definition, Uses

What is an Inverted Microscope? An inverted microscope is a type of microscope that is designed with the objective lenses facing downward and the stage ...

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What is an Inverted Microscope?

An inverted microscope is a type of microscope that is designed with the objective lenses facing downward and the stage (where the specimen is placed) facing upward. In this configuration, the viewer looks down into the microscope to observe the specimen, rather than looking through the eyepieces as with a traditional compound microscope. Inverted microscopes are used in a variety of applications, including cell culture, tissue culture, and live-cell imaging.

One advantage of an inverted microscope is that it allows the user to observe large, flat specimens or cell cultures without having to move the specimen or culture dish. This is particularly useful for studying living cells, as it allows the cells to remain in their natural environment while being observed. Inverted microscopes are also useful for observing samples that are too large or too heavy to be mounted on a traditional microscope slide.

Inverted microscopes are often equipped with a number of specialized features to facilitate their use, such as temperature-controlled stages, special illumination systems, and motorized stages for moving the specimen. These features can add to the cost of the microscope, but they can also be essential for certain applications.

The inverted microscope was first developed in the late 19th century as a way to study large, flat specimens such as slides and petri dishes. Prior to the development of the inverted microscope, scientists were limited to studying small specimens that could be mounted on traditional microscope slides.

The first inverted microscope was developed by Carl Zeiss in 1872, and it was known as the “stand microscope.” This instrument was designed with the objective lenses facing downward and the stage facing upward, similar to modern inverted microscopes. In the following years, other manufacturers developed their own versions of the inverted microscope, and the design became widely used in scientific research and industrial applications.

Over time, the inverted microscope has undergone numerous improvements and refinements. Today, inverted microscopes are widely used in a variety of applications, including cell culture, tissue culture, and live-cell imaging. They are also used in industrial and manufacturing settings to inspect parts and components.

Features of Inverted Microscope

Inverted microscopes typically have a number of specialized features that are designed to facilitate their use:

  1. Motorized stage: An inverted microscope may have a motorized stage, which allows the user to move the specimen or culture dish in a precise and controlled manner. This can be particularly useful when studying large or complex specimens.
  2. Specialized illumination systems: Inverted microscopes may have specialized illumination systems, such as LED lighting or fluorescent lighting, to provide a bright, even illumination of the specimen. These systems may also have adjustable intensity and color temperature to allow for different lighting conditions.
  3. Temperature-controlled stage: Some inverted microscopes have a temperature-controlled stage, which allows the user to maintain a stable temperature for the specimen or culture dish. This can be particularly useful when studying living cells, as it allows the cells to remain in their optimal temperature range.
  4. Phase contrast or DIC optics: Inverted microscopes may also have phase contrast or differential interference contrast (DIC) optics, which allow the user to view fine details and structures within the specimen that might not be visible with traditional brightfield illumination.
  5. Fluorescence imaging capabilities: Some inverted microscopes are equipped with fluorescence imaging capabilities, which allow the user to study samples that have been labeled with fluorescent dyes or proteins. This can be particularly useful for studying the distribution and movement of molecules within cells or tissues.
  6. High-resolution cameras: Many inverted microscopes are equipped with high-resolution cameras, which allow the user to capture images or video of the specimen for analysis or documentation. Some cameras may also have specialized features, such as time-lapse capability or the ability to capture multiple wavelengths of light.

Types of Inverted microscope

There are several different types of inverted microscopes, each designed for specific applications:

  1. Basic inverted microscopes: These are the most basic type of inverted microscopes and are typically used for routine observations of cultures and slides. They may have simple illumination systems and fixed stages, and may not have advanced features such as motorized stages or fluorescence imaging capabilities.
  2. Advanced inverted microscopes: These microscopes are designed for more specialized applications and may have a wider range of features and capabilities. They may have motorized stages, advanced illumination systems, and fluorescence imaging capabilities, as well as other specialized features such as phase contrast or DIC optics.
  3. Live-cell imaging microscopes: These microscopes are designed specifically for studying living cells and tissues and are equipped with features such as temperature-controlled stages, specialized illumination systems, and fluorescence imaging capabilities. They may also have specialized features such as oxygen control or CO2 gas control to maintain a stable environment for the cells.
  4. Industrial inverted microscopes: These microscopes are designed for use in industrial and manufacturing settings and are used to inspect parts and components. They may have specialized features such as high-resolution cameras, motorized stages, and specialized illumination systems to facilitate the inspection process.

Principle of Inverted Microscope

Both the upright light microscope and the inverted microscope operate on the same fundamental basis. The objective lenses create a focused image of the specimen using light rays. However, the light source and condenser in an inverted microscope are located above the stage and directed downward. Above the specimen stage is a condenser lens, whose primary purpose is to focus light directly onto the specimen below. The specimen is displayed on a stage that is spacious enough to accommodate it. Light from the condenser is collected by the objectives positioned below the stage and directed upwards, where the picture is magnified before being transmitted to the ocular lens. The ocular lens acts like a mirror, reflecting light into the eye. Through the use of an ibidi Polymer Coverslip and an ibidi Glass coverslip, the cells can be examined and observed through the base of the cell culture apparatus, where the maximum number of optical points is attained.

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;

inverted microscope labeled
inverted microscope labeled

The main parts of an inverted microscope include:

  1. Objective lenses: These lenses are located at the bottom of the microscope and are used to collect light from the specimen. Inverted microscopes typically have several objective lenses of different magnifications, allowing the user to choose the appropriate magnification for the specimen being studied.
  2. Stage: The stage is the flat surface where the specimen or culture dish is placed. It is located at the top of the microscope and is usually adjustable to allow the user to position the specimen in the optimal location for viewing.
  3. Stage Clips: It holds the specimen in position.
  4. Arm: It holds the optical and mechanical parts of the microscope.
  5. Illumination system: The illumination system provides light to the specimen and may consist of a light source, such as an LED or fluorescent lamp, and a series of lenses and filters to direct the light onto the specimen.
  6. Condenser lens: The condenser lens is located below the stage and is used to focus the light onto the specimen. It may have a diaphragm to control the amount of light that reaches the specimen.
  7. Nosepiece: It holds the objective lens.
  8. Focus knobs: Inverted microscopes typically have two focus knobs, one for coarse focus and one for fine focus, which allow the user to adjust the focus of the microscope.
  9. Eyepiece: The eyepiece is located at the top of the microscope and is used to view the specimen. Inverted microscopes may have one or more eyepieces, depending on the model.
  10. Control panel: Many inverted microscopes have a control panel, which allows the user to adjust the various settings and features of the microscope. This may include controls for the stage, illumination system, and other features such as fluorescence imaging or time-lapse capabilities.
  11. The Dual concentric knobs: It contains fine and coarse adjustment knobs which helps in fine-tuning and focusing the objectives to the specimen.
  12. Digital Camera: It used to record or capture the image of the specimen.

Operating Procedure of Inverted Microscope

  1. Placed the inverted microscope on a table in stable condition.
  2. Then placed the specimen containing slide or glass container on the specimen stage.
  3. The specimen stage does not move at all.
  4. Now adjust the focus by moving the objective and condenser lens (use the knobs to control the movement of Condenser and Objective Lens).
  5. The image of the specimen can be observed by the ocular lens or on a digital screen.
  6. 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.

Inverted microscope uses

Inverted microscopes are used in a variety of applications, including:

  1. Cell culture: Inverted microscopes are commonly used in cell culture applications to study living cells in their natural environment. They allow the user to observe cells as they grow and divide, and can be used to study the effects of various treatments or conditions on cell behavior. 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.
  2. Tissue culture: Inverted microscopes are also commonly used in tissue culture applications to study the behavior of cells in tissue samples. They allow the user to observe the interactions between cells and the surrounding environment, and can be used to study the effects of various treatments or conditions on tissue behavior.
  3. Live-cell imaging: Inverted microscopes are often equipped with specialized features such as temperature-controlled stages and fluorescence imaging capabilities, which make them ideal for studying living cells and tissues in real-time.
  4. Industrial inspection: Inverted microscopes are also used in industrial and manufacturing settings to inspect parts and components for defects or abnormalities. They may have specialized features such as high-resolution cameras and motorized stages to facilitate the inspection process.
  5. Scientific research: Inverted microscopes are widely used in scientific research to study a variety of biological and chemical phenomena. They may be used to study the behavior of cells, tissues, and other small structures, and can be used to study the effects of various treatments or conditions on these structures.

Advantages of Inverted Microscopy

There are several advantages to using an inverted microscope:

  1. Allows observation of large, flat specimens: One of the main advantages of an inverted microscope is that it allows the user to observe large, flat specimens or cell cultures without having to move the specimen or culture dish. This is particularly useful for studying living cells, as it allows the cells to remain in their natural environment while being observed.
  2. Allows for easy manipulation of the specimen: Because the stage of an inverted microscope is located at the top of the microscope, the user can easily manipulate the specimen or culture dish without having to reach into the microscope. This can be particularly useful when working with large or delicate specimens.
  3. Allows for the use of specialized illumination systems: Inverted microscopes may have specialized illumination systems, such as LED or fluorescent lighting, which can provide a bright, even illumination of the specimen. This can be particularly useful for studying living cells or other transparent specimens.
  4. Allows for the use of specialized optics: Inverted microscopes may also have specialized optics, such as phase contrast or differential interference contrast (DIC) optics, which allow the user to view fine details and structures within the specimen that might not be visible with traditional brightfield illumination.
  5. Allows for the use of specialized cameras: Many inverted microscopes are equipped with high-resolution cameras, which allow the user to capture images or video of the specimen for analysis or documentation. This can be particularly useful for studying the behavior of cells or tissues over time.
  6. Living cells: Due to the inverted microscope’s large viewing area, it is frequently employed for studying living cells by peering at them from below in a Petri dish or glass tube.
  7. Culture Media: Additionally, it can be utilised to see and analyse cells in a substantial quantity of media.
  8. Field of vision: In comparison to the traditional upright microscope, which can only examine specimens on tiny microscopic slides, this one allows you to observe cell tissues still in their original vessel, which greatly expands your field of vision.
  9. Enormous volumes of the media: In contrast to viewing cells as microscopic specimens on a glass slide under a coverslip, it is possible to observe cells in enormous volumes of the media.
  10. Observation of the cells in petri Plate: Because most cells will sink to the bottom of the holding equipment, such as a Petri plate, observation of the cells is possible from both the top and bottom of the plate.
  11. Cleanliness: Maintaining the sample’s cleanliness is made possible since the objective lens never comes into contact with the specimen.

Disadvantages of Inverted Microscopy

There are a few disadvantages to using an inverted microscope:

  1. Limited magnification: Inverted microscopes typically have lower magnifications compared to upright microscopes, which can make it more difficult to study small structures or details.
  2. Limited depth of field: Inverted microscopes also tend to have a limited depth of field, meaning that only objects at a specific distance from the lens are in focus at any given time. This can make it more difficult to study specimens with a range of depths, such as thick tissue samples.
  3. More expensive: Inverted microscopes are often more expensive than upright microscopes, particularly when equipped with specialized features such as temperature-controlled stages or fluorescence imaging capabilities.
  4. Requires specialized training: Using an inverted microscope may require more specialized training compared to an upright microscope, as the user must be familiar with the specific features and capabilities of the instrument.
  5. Limited to studying flat specimens: Inverted microscopes are limited to studying flat specimens or cell cultures, and are not suitable for studying small specimens that are mounted on traditional microscope slides.

What is the difference between an inverted versus an upright microscope?

The main difference between an inverted microscope and an upright microscope is the orientation of the objective lenses and the stage. In an inverted microscope, the objective lenses are located at the bottom of the microscope and the stage is located at the top. This allows the user to observe large, flat specimens or cell cultures without having to move the specimen or culture dish. In contrast, an upright microscope has the objective lenses located at the top of the microscope and the stage located at the bottom. This configuration is more suited to studying small specimens that are mounted on traditional microscope slides.

Other differences between inverted and upright microscopes include the type of illumination used and the type of image that is produced. Inverted microscopes typically use transmitted light, which passes through the specimen and is then focused by the lenses. This can be particularly useful for studying living cells, as it allows the cells to remain in their natural environment while being observed. In contrast, upright microscopes may use either transmitted or reflected light, depending on the type of specimen being studied.

In terms of the image produced, inverted microscopes typically produce a stereo image, which allows the user to view the specimen in three dimensions. In contrast, upright microscopes produce a flat image that is viewed through the eyepieces.

Inverted epifluorescence microscope

An inverted epifluorescence microscope is a type of inverted microscope that is equipped with fluorescence imaging capabilities. Fluorescence microscopy is a technique in which samples are labeled with fluorescent dyes or proteins, which emit light when excited by a specific wavelength of light. This allows the user to study the distribution and movement of molecules within cells or tissues.

An inverted epifluorescence microscope typically has two light sources: a transmitted light source and an epifluorescence light source. The transmitted light source is used to illuminate the specimen in the traditional manner, while the epifluorescence light source is used to excite the fluorescent dyes or proteins within the specimen. The emitted fluorescence is then collected by the objective lenses and directed to the eyepieces, where it is visible to the user.

Inverted epifluorescence microscopes may also have specialized filters and dichroic mirrors to separate the transmitted light and the emitted fluorescence, and to allow the user to visualize specific wavelengths of light. These features can be particularly useful for studying multiple fluorophores within the same sample.

What is inverted metallurgical microscope?

An inverted metallurgical microscope is a type of inverted microscope that is specifically designed for studying metallic materials. It is equipped with specialized features such as polarizing filters, phase contrast optics, and specialized illumination systems that allow the user to visualize the microstructure of metallic materials in detail.

Inverted metallurgical microscopes are commonly used in a variety of industries, including aerospace, automotive, and materials science, to study the properties and behavior of metallic materials. They can be used to identify defects or abnormalities in materials, to study the effects of various treatments or conditions on materials, and to analyze the microstructure of materials at the microscopic scale.

Inverted metallurgical microscopes are typically more expensive than traditional inverted microscopes, due to the specialized features and capabilities that they offer. They may also require more specialized training to use effectively.

Examples of Inverted Microscopy

Leica inverted microscope

Leica is a leading manufacturer of scientific instruments, including inverted microscopes. Leica inverted microscopes are designed for a variety of applications, including cell culture, tissue culture, live-cell imaging, and scientific research. They are known for their high-quality optics and advanced features, such as motorized stages, temperature-controlled stages, and fluorescence imaging capabilities.

Leica inverted microscopes are available in a range of models to suit different applications and budgets. Basic models may be suitable for routine observations of cultures and slides, while more advanced models may have a wider range of features and capabilities for specialized applications.

Leica inverted microscopes are widely used in scientific research and medical settings, as well as in industrial and manufacturing settings. They are known for their reliability and durability, and are backed by Leica’s reputation for quality and innovation.

Olympus inverted microscope

Olympus is a leading manufacturer of scientific instruments, including inverted microscopes. Olympus inverted microscopes are designed for a variety of applications, including cell culture, tissue culture, live-cell imaging, and scientific research. They are known for their high-quality optics and advanced features, such as motorized stages, temperature-controlled stages, and fluorescence imaging capabilities.

Olympus inverted microscopes are available in a range of models to suit different applications and budgets. Basic models may be suitable for routine observations of cultures and slides, while more advanced models may have a wider range of features and capabilities for specialized applications.

Olympus inverted microscopes are widely used in scientific research and medical settings, as well as in industrial and manufacturing settings. They are known for their reliability and durability, and are backed by Olympus’s reputation for quality and innovation.

Nikon inverted microscope

Nikon is a leading manufacturer of scientific instruments, including inverted microscopes. Nikon inverted microscopes are designed for a variety of applications, including cell culture, tissue culture, live-cell imaging, and scientific research. They are known for their high-quality optics and advanced features, such as motorized stages, temperature-controlled stages, and fluorescence imaging capabilities.

Nikon inverted microscopes are available in a range of models to suit different applications and budgets. Basic models may be suitable for routine observations of cultures and slides, while more advanced models may have a wider range of features and capabilities for specialized applications.

Nikon inverted microscopes are widely used in scientific research and medical settings, as well as in industrial and manufacturing settings. They are known for their reliability and durability, and are backed by Nikon’s reputation for quality and innovation.

Inverted microscope price

The price of an inverted microscope can vary widely depending on the features and capabilities of the instrument. Basic inverted microscopes may start at around $1,000, while more advanced models with specialized features such as motorized stages, temperature-controlled stages, and fluorescence imaging capabilities can cost upwards of $10,000 or more.

Factors that can influence the price of an inverted microscope include the manufacturer, the quality of the optics, the number and type of objective lenses, the type of illumination system, and the presence of specialized features such as phase contrast optics or fluorescence imaging capabilities.

It is worth noting that inverted microscopes are typically more expensive than traditional upright microscopes, due to the specialized features and capabilities that they offer. However, for certain applications, such as studying living cells or tissues, the benefits of an inverted microscope may justify the higher cost.

Inverted Microscope Free Worksheet

Inverted Microscope Free Worksheet
Inverted Microscope Free Worksheet

Inverted microscope Images

Inverted microscope Images
Inverted microscope Images – Cell Culture

Reference

  • https://en.wikipedia.org/wiki/Inverted_microscope
  • https://microbenotes.com/inverted-microscope/
  • https://www.microscopemaster.com/inverted-microscope.html
  • https://ibidi.com/content/212-inverted-and-upright-microscopy
  • https://www.biocompare.com/25969-Inverted-Microscopes/
  • https://www.microscopeworld.com/t-inverted_microscopes.aspx

Citation

APA

MN Editors. (July 17, 2020).Inverted Microscope – Principle, Parts, Definition, Uses. Retrieved from https://microbiologynote.com/inverted-microscope-principle-parts-definition-uses/

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MN Editors. "Inverted Microscope – Principle, Parts, Definition, Uses." Microbiology Note, Microbiologynote.com, July 17, 2020.

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