Name: how microscopes reach higher resolution
Uploaded: 2023-06-23T15:38:43.000Z
Duration: 1 min 22 s
Channel: JoVE

K12 Standards

Virginia  Standards of Learning

HIgh School

Life Science

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LS.1.b

Overview of Microscopy Techniques

Microscopy is a technique that uses microscopes to magnify tiny cellular details and study tissue characteristics.
Based on the principle of generating images, microscopes can be of different types.
Optical microscopy uses light to visualize samples and magnify objects up to 1000 times. The compound microscope is the most commonly used optical microscope.
Electron microscopy uses an electron beam instead of light, allowing 2 to 50 million times magnification.
Depending on how the electron beam interacts with the sample, there are two major types of electron microscopy.
Transmission electron microscopy can generate 2D images providing information about a sample&#39;s internal structure and composition. While scanning electron microscopy provides information about the 3D topography and surface composition of samples like bone and hair.
In contrast, atomic force microscopy or AFM uses a scanning probe that follows the contours of the sample surface and can capture a 3D image of the sample surface. AFM can achieve up to 1 million times magnification.

overview of microscopy techniques

The early pioneers of microscopy opened a window into the invisible world of microorganisms. In 1830, Joseph Jackson Lister created an essentially modern light microscope. The 20th century saw the development of microscopes that leveraged nonvisible light, such as fluorescence microscopy that uses an ultraviolet light source and electron microscopy that uses short-wavelength electron beams. These advances significantly improved magnification, image resolution, and contrast. By comparison, the relatively rudimentary microscopes of van Leeuwenhoek and his contemporaries were far less powerful than even the most basic microscopes in use today.
Light Microscopy
Many types of microscopes fall under the category of light microscopes, which use light to visualize images. Examples of light microscopes include brightfield microscopes, darkfield microscopes, phase-contrast microscopes, differential interference contrast microscopes, fluorescence microscopes, confocal scanning laser microscopes, and two-photon microscopes. These various types of light microscopes can complement each other in diagnostics and research.
<h3>Electron Microscopy</h3>
The wavelengths of visible light limit the maximum theoretical resolution of images created by light microscopes. Most light microscopes can only magnify 1000x, and a few can magnify up to 1500x. However, this does not begin to approach the magnifying power of an electron microscope (EM), which uses short-wavelength electron beams rather than light to increase magnification and resolution.
Electrons, like electromagnetic radiation, can behave as waves, but with wavelengths of 0.005 nm, they can produce much better resolution than visible light. An EM can produce a sharp image that is magnified up to 100,000x. EMs can resolve subcellular structures and some molecular structures (e.g., single strands of DNA); however, electron microscopy cannot be used on living material because of the methods needed to prepare the specimens.
There are two basic types of EM: the transmission electron microscope (TEM) and the scanning electron microscope (SEM). The TEM is somewhat analogous to the brightfield light microscope in terms of the way it functions. However, it uses an electron beam from above the specimen that is focused using a magnetic lens (rather than a glass lens) and projected through the specimen onto a detector. Electrons pass through the specimen, and then the detector captures the image.
Scanning Probe Microscopy
A scanning probe microscope does not use light or electrons but rather very sharp probes that are passed over the surface of the specimen and interact with it directly. This produces information that can be assembled into images with magnifications up to 100,000,000x. Such large magnifications can be used to observe individual atoms on surfaces. To date, these techniques have been used primarily for research rather than for diagnostics. There are two types of scanning probe microscopes: the scanning tunneling microscope (STM) and the atomic force microscope (AFM).
This text is adapted from <a href="https://openstax.org/books/microbiology/pages/2-3-instruments-of-microscopy">Openstax, Microbiology, Section 2.3: Instruments of Microscopy</a>

The early pioneers of microscopy opened a window into the invisible world of microorganisms. In 1830, Joseph Jackson Lister created an essentially modern ...

Education

JoVE Core

Anatomy and Physiology

Unit 1

Diagnostic Imaging Techniques

KEY TERMS AND CONCEPTS

how microscopes reach higher resolution

Microscopes have become more powerful by improving magnification, resolution, and contrast. Early pioneers such as van Leeuwenhoek used far simpler instruments than the microscopes used today. In 1830, Joseph Jackson Lister created an essentially modern light microscope. Later advances in the 20th century added nonvisible light, including fluorescence microscopy and electron microscopy.

Many microscopes used in schools and labs are light microscopes. These instruments use light to make images and include brightfield, darkfield, phase-contrast, differential interference contrast, fluorescence, confocal scanning laser, and two-photon microscopes. These types can work together in diagnostics and research. Each one helps highlight different details in a specimen.

Visible light limits how much a light microscope can resolve. Most light microscopes magnify up to 1000x, and a few reach 1500x. Electron microscopes use short-wavelength electron beams instead of light, so they can achieve much higher resolution. Because electrons have wavelengths of about 0.005 nm, an electron microscope can magnify up to 100,000x and reveal subcellular structures and some molecular structures, including single strands of DNA. Electron microscopy cannot be used on living material because of the specimen preparation methods.

There are two basic types of electron microscopes: the transmission electron microscope, or TEM, and the scanning electron microscope, or SEM. The TEM works somewhat like a brightfield light microscope, but it uses an electron beam from above the specimen and magnetic lenses instead of glass lenses. The beam passes through the specimen, and a detector captures the image. Scanning probe microscopy uses a different approach. It does not use light or electrons. Instead, very sharp probes move over the specimen surface and interact with it directly.

Scanning probe microscopes can build images at magnifications up to 100,000,000x. At that scale, individual atoms on surfaces can be observed. The two main types are the scanning tunneling microscope, or STM, and the atomic force microscope, or AFM. These methods are used mainly for research rather than diagnostics.

How Microscopes Reach Higher Resolution

x-ray imaging

X-Ray Use in Medical Diagnosis

X-rays are a standard tool in medical diagnosis. They help doctors see bones, teeth, and some soft tissues without surgery. The first X-ray image showed Wilhelm Röntgen’s wife’s hand, and it opened a new way to look inside the body. Wilhelm Röntgen, a German physicist, discovered X-rays in 1895 while studying electrical current. He found that an invisible ray could pass through flesh but leave the bones outlined on a screen coated with a metal compound. His work led to the first durable record...

computed tomography

CT Scans for Body Slices

Computed tomography, or CT, creates detailed body slices using X-rays and computers. It is a non-invasive imaging method that helps reveal fine details inside the body. The word tomography means imaging by sections. CT was developed in the 1970s. It works by measuring how X-rays pass through the body and are absorbed or reflected at different levels. During a scan, the patient lies on a motorized platform while a computerized axial tomography, or CAT, scanner rotates 360 degrees around the...

magnetic resonance imaging

MRI Scans: How Body Signals Are Formed

Magnetic resonance imaging, or MRI, is a noninvasive way to create detailed medical images. It uses strong magnetic fields and radio waves. When the body is exposed to them, it can emit radio signals that are used to build the image. MRI became especially important after researchers saw that cancerous tissue gave off signals different from normal tissue. Raymond Damadian identified this difference in 1970 and later applied for a patent for the first MRI scanner used in clinical care. Early...

positron emission tomography

PET Scans and How They Work

PET scans use radiopharmaceuticals to show how the body is working. PET, or positron emission tomography, is a medical imaging method that depends on short-lived radiation from these compounds. It gives doctors a way to see tissue activity instead of only body structure. A PET scan starts with a positron-emitting radioisotope. This isotope is made in a cyclotron and then attached to a substance used by the body part being studied. The tagged compound is called a radiotracer. It can be given by...

ultrasonography

Ultrasonography in Medical Imaging

Ultrasonography in medical imaging uses high-frequency sound waves to create images of internal body structures. It is a safe, non-invasive procedure. It does not use ionizing radiation, which is one reason it is widely used in medicine. During an ultrasound exam, a handheld transducer is placed on the skin. A transducer is the device that sends sound waves into the body. When the waves hit different tissues, they bounce back as echoes. The transducer catches those echoes and sends them to a...

Microscopes have become more powerful by improving magnification, resolution, and contrast. Early pioneers such as van Leeuwenhoek used far simpler instruments than the microscopes used today. In 1830, Joseph Jackson Lister created an essentially modern light microscope. Later advances in the 20th century added nonvisible light, including fluorescence microscopy and electron microscopy. Many microscopes used in schools and labs are light microscopes. These instruments use light to make images...

key terms and concepts

Learn how light microscopes, electron microscopes, and scanning probe microscopes differ in magnification, resolution, and what they can reveal.

How Microscopes Reach Higher Resolution | JoVE K12

anatomy and physiology

JoVE Core Anatomy and Physiology showcases the anatomical and physiological details of the human body in comprehensive, clear, and engaging video animations. This collection of lessons focuses on the structures, components, functions, and requirements of the human body. It is complemented by scientist-in-action videos that demonstrate the application of this knowledge in real-world medical and research settings.

Next Generation Science Standards

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Physical Science

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Waves and Their Applications in Technologies for Information Transfer

HS-PS4-5

Florida Department of Education

SC.912.L.14

SC.912.L.14.4

new-york-state-science-learning-standards

New York State Science Learning Standards

HS. Waves and Electromagnetic Radiation

HS-PS4-5.

Scientific Method

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Scientific Laws and Theories

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• ask questions and develop hypotheses to determine relationships between independent and dependent variables
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LS.1.a

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Introduction to Light Microscopy

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The student will demonstrate an understanding of scientific and engineering practices by planning and carrying out investigations
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Scatter Plot

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Bar Graph

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Reasoning ...

The student will demonstrate an understanding of scientific and engineering practices by constructing and critiquing conclusions and explanations
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The student will demonstrate an understanding of scientific and engineering practices by obtaining, evaluating, and communicating information
• read scientific texts, including those adapted for classroom use, to obtain scientific and/or technical information
• gather, read, and synthesize information from multiple appropriate sources and assess the credibility, accuracy, and possible bias of each publication
• construct, use, and/or present an argument supported by empirical evidence and scientific reasoning

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How Microscopes Reach Higher Resolution

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