Introducation to Microscopy
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In this presentation, I have described the Principle and Application of Light, Phase contrast, Fluorescence.
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Phase contrast microscopy
The document discusses different light microscopy techniques including dark field microscopy, phase contrast microscopy, and fluorescence microscopy. Dark field microscopy uses angled illumination to visualize objects that scatter light against a dark background. Phase contrast microscopy converts small phase changes in light passing through a specimen into visible intensity changes, allowing fine specimen details to be seen. Fluorescence microscopy takes advantage of specimens that are inherently fluorescent or can be labeled with fluorescent dyes. These techniques allow better visualization of transparent or unstained specimens compared to traditional bright field microscopy.
phase contrast microscope
The document discusses phase contrast microscopy, which was developed by Fritz Zernike in the 1930s. It allows living or unstained cells and intracellular components to be visible under a microscope. The phase contrast microscope works by converting small phase changes caused by differences in refractive index of cell structures into visible brightness and darkness differences. This makes organelles and other structures visible without using staining. The phase contrast is achieved using an annular diaphragm and phase rings or filters to shift the phase of light passing through or around the specimen.
Microscopy
Microscopy uses lenses to magnify objects too small to see with the naked eye. A microscope contains objective lenses close to the sample that collect light and project an enlarged virtual image, and eyepiece lenses that further magnify the image for viewing. Key components include the condenser, which directs light up through the sample, and various types of objectives and eyepieces optimized for tasks like brightfield, darkfield, phase contrast, or fluorescence microscopy. Objectives are classified by their magnification and degree of correction for chromatic and spherical aberration, from simple achromats to highly-corrected apochromats. Different microscopy techniques employ specific lighting and optics to reveal features of transparent and unstained samples.
Simple microscope
The document discusses the simple microscope. It defines a simple microscope as using a single lens for magnification rather than multiple lenses like a compound microscope. A simple microscope works by using a convex lens to produce a virtual, erect and magnified image of an object placed within the lens' focal point. The maximum magnification of a simple microscope is around 10x. Key parts of a simple microscope include a metal stand, stage to hold samples, plano-convex mirror to illuminate samples, and a biconvex lens for magnification. Simple microscopes are used to examine small biological specimens, watch parts, jewelry, book text, soil particles, and skin.
Phase contrast microscope
Phase-contrast microscopy is a technique that converts phase shifts in light passing through a transparent specimen to brightness changes in the image, allowing living cells that are otherwise invisible to be seen. It works by separating light rays that pass through a specimen unchanged from those that are diffracted, using an annular diaphragm and phase plate in the light path. Phase-contrast microscopy is widely used in biological research for observing living cells, microorganisms, and other transparent specimens without staining or fixing.
Light microscopy
this presentation deals with the introduction of some of the commonly used optical microscopes in forensic labs; compound microscope, stereoscopic microscope, comparison microscope, fluorescence microscope and polarized microscope.
PHASE MICROSCOPY
This document discusses phase contrast microscopy. It begins by defining a microscope and microscopy. It then describes the main types of microscopes, including optical, electron, and scanning probe microscopes. It focuses on the light microscope, explaining that it uses visible light and is commonly used in biology to view structures like cells. It defines magnification and resolution. It describes the different modes of light microscopy - bright field, dark field, and phase contrast - and explains that phase contrast microscopy allows observation of living cells by enhancing contrast between structures with small refractive index differences. It notes that phase contrast microscopy was invented by Frits Zernike, for which he won the Nobel Prize in Physics in 1953.
Phase contrast microscope
Phase contrast microscopy is a technique that was invented in 1934 by Dutch physicist Frits Zernike, for which he received the Nobel Prize in Physics in 1953. It allows for high-contrast imaging of transparent specimens like living cells without staining. In phase contrast microscopy, variations in the refractive index within a specimen cause some light rays to be retarded, producing an image with areas of different intensities. This converts subtle differences in a sample's density and refractive index into detectable variations in light intensity. Phase contrast microscopy is useful for observing living cells and intracellular structures in their natural state without needing to kill, fix, or stain the specimen. While it provides high resolution living images, it has limitations such as inability to view thick specimens clearly and
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Phase contrast microscopy
The document discusses different light microscopy techniques including dark field microscopy, phase contrast microscopy, and fluorescence microscopy. Dark field microscopy uses angled illumination to visualize objects that scatter light against a dark background. Phase contrast microscopy converts small phase changes in light passing through a specimen into visible intensity changes, allowing fine specimen details to be seen. Fluorescence microscopy takes advantage of specimens that are inherently fluorescent or can be labeled with fluorescent dyes. These techniques allow better visualization of transparent or unstained specimens compared to traditional bright field microscopy.
phase contrast microscope
The document discusses phase contrast microscopy, which was developed by Fritz Zernike in the 1930s. It allows living or unstained cells and intracellular components to be visible under a microscope. The phase contrast microscope works by converting small phase changes caused by differences in refractive index of cell structures into visible brightness and darkness differences. This makes organelles and other structures visible without using staining. The phase contrast is achieved using an annular diaphragm and phase rings or filters to shift the phase of light passing through or around the specimen.
Microscopy
Microscopy uses lenses to magnify objects too small to see with the naked eye. A microscope contains objective lenses close to the sample that collect light and project an enlarged virtual image, and eyepiece lenses that further magnify the image for viewing. Key components include the condenser, which directs light up through the sample, and various types of objectives and eyepieces optimized for tasks like brightfield, darkfield, phase contrast, or fluorescence microscopy. Objectives are classified by their magnification and degree of correction for chromatic and spherical aberration, from simple achromats to highly-corrected apochromats. Different microscopy techniques employ specific lighting and optics to reveal features of transparent and unstained samples.
Simple microscope
The document discusses the simple microscope. It defines a simple microscope as using a single lens for magnification rather than multiple lenses like a compound microscope. A simple microscope works by using a convex lens to produce a virtual, erect and magnified image of an object placed within the lens' focal point. The maximum magnification of a simple microscope is around 10x. Key parts of a simple microscope include a metal stand, stage to hold samples, plano-convex mirror to illuminate samples, and a biconvex lens for magnification. Simple microscopes are used to examine small biological specimens, watch parts, jewelry, book text, soil particles, and skin.
Phase contrast microscope
Phase-contrast microscopy is a technique that converts phase shifts in light passing through a transparent specimen to brightness changes in the image, allowing living cells that are otherwise invisible to be seen. It works by separating light rays that pass through a specimen unchanged from those that are diffracted, using an annular diaphragm and phase plate in the light path. Phase-contrast microscopy is widely used in biological research for observing living cells, microorganisms, and other transparent specimens without staining or fixing.
Light microscopy
this presentation deals with the introduction of some of the commonly used optical microscopes in forensic labs; compound microscope, stereoscopic microscope, comparison microscope, fluorescence microscope and polarized microscope.
PHASE MICROSCOPY
This document discusses phase contrast microscopy. It begins by defining a microscope and microscopy. It then describes the main types of microscopes, including optical, electron, and scanning probe microscopes. It focuses on the light microscope, explaining that it uses visible light and is commonly used in biology to view structures like cells. It defines magnification and resolution. It describes the different modes of light microscopy - bright field, dark field, and phase contrast - and explains that phase contrast microscopy allows observation of living cells by enhancing contrast between structures with small refractive index differences. It notes that phase contrast microscopy was invented by Frits Zernike, for which he won the Nobel Prize in Physics in 1953.
Phase contrast microscope
Phase contrast microscopy is a technique that was invented in 1934 by Dutch physicist Frits Zernike, for which he received the Nobel Prize in Physics in 1953. It allows for high-contrast imaging of transparent specimens like living cells without staining. In phase contrast microscopy, variations in the refractive index within a specimen cause some light rays to be retarded, producing an image with areas of different intensities. This converts subtle differences in a sample's density and refractive index into detectable variations in light intensity. Phase contrast microscopy is useful for observing living cells and intracellular structures in their natural state without needing to kill, fix, or stain the specimen. While it provides high resolution living images, it has limitations such as inability to view thick specimens clearly and
Microscopy
The microscope has evolved a lot from the time of Leeuwenhoek. This presentation gives a brief overview about the types of microscope their principle of function and application.
Bright field microscope and its working principle
The document discusses the bright field microscope. It notes that the bright field microscope is a simple microscope to use, with few adjustments needed. It can view some specimens without staining and does not alter specimen color. However, it has limitations such as distortion with high contrast and inability to view living bacteria. The document describes how bright field microscopy works by using a light source to illuminate the specimen from below, appearing dark against a bright background, and provides examples of specimens that can be observed with it.
Electron microscope
The document discusses the history and development of electron microscopes. It describes how J.J. Thomson discovered electrons in 1897 and how subsequent scientists like de Broglie, Ruska, and Knoll contributed to the development of the first electron microscope in the 1930s. It then explains the basic workings and components of transmission electron microscopes and scanning electron microscopes, how they produce images, and some examples of their applications in biology and materials science.
Light Microscopy
The document provides information about using a compound light microscope, including:
1) The main parts of a compound light microscope are the ocular lenses, body tube, objective lenses, stage, condenser, iris diaphragm, coarse and fine adjustment knobs, and light source.
2) To examine a specimen, the user selects the appropriate objective lens, focuses using the coarse and fine adjustment knobs, and adjusts the condenser and iris diaphragm to control illumination.
3) For oil immersion objectives, oil is added to the slide and specimen to allow for higher magnification imaging.
Microscope ppt, by jitendra kumar pandey,medical micro,2nd yr, mgm medical co...
The document summarizes different types of microscopes used to study microorganisms. It discusses light microscopes like brightfield, darkfield and phase contrast microscopes. It also describes electron microscopes like transmission electron microscopes (TEM) and scanning electron microscopes (SEM) that use electron beams instead of light. TEM images internal structures by transmitting electrons through thin samples while SEM scans sample surfaces using secondary electrons. Sample preparation methods for both light and electron microscopy are also outlined.
Microscope and its types
This document provides information about different types of microscopes. It discusses simple microscopes, which use a single lens for magnification, and their basic parts and working principle. Compound microscopes are then introduced as using multiple lenses to achieve higher magnification than is possible with a simple lens alone. The key parts of a compound microscope are described, including the eyepiece, objective lenses, body tube, arm, and focus adjustments. It explains that light from the illuminator passes through these parts to produce a magnified image for viewing.
Different types of microscopes
This document provides information on different types of microscopy techniques including bright field, dark field, phase contrast, and polarized light microscopy. It begins with explaining the basics of light and microscopy. It then describes each technique in more detail, including their principles, applications, advantages, and how they are set up optically. Bright field microscopy uses illumination and forms a dark image on a bright background. Dark field uses oblique illumination to see small particles as bright objects on a dark background. Phase contrast converts phase differences into contrast changes to see transparent specimens. Polarized light microscopy uses polarized filters to reveal structural details not otherwise seen.
Electron microscope (TEM & SEM)
TEM and SEM are two different Electron microscope used to view objects which otherwise are impossible to see under normal light/ compound microscope.
Dark field microscope
Dark field microscopy produces bright images of unstained samples against a dark background. It works by using a condenser with an opaque disk to block light entering the objective lens directly, allowing only light reflected off the sample to pass through. This causes specimens to appear bright on a dark background. It is useful for viewing transparent or unstained samples like bacteria, cells, and minerals due to the contrast it provides.
Light phase contrast and fluorescence microscopy
Phase contrast microscopy is a type of light microscopy technique that produces high contrast images of transparent specimens without staining. It works by interfering surrounding light waves and sample-diffracted waves to create an image with the inner regions of the sample appearing darker than the outer regions. Fluorescence microscopy uses fluorescent dyes excited by high-energy light to emit lower-energy light, allowing specific structures within samples to be imaged. It is used to study living cells and view genetic material.
1. MICROSCOPY - introduction + principle (Basics)
Basics only
Microscopy is the technical field that uses microscopes to observe samples which are
not in the resolution range of the normal-unaided eye.
Microscope is a scientific-instrument consisting of magnifying lens that enables an
observer to view the minute features distinctly.
In greek, micro = small
skopein = to view.
Recent advancement in microscopic techniques cryoem and super
Cryo-electron microscopy is an emerging technique that uses electron microscopes to image biomolecules such as proteins that have been rapidly frozen to preserve their structure. It overcomes limitations of other techniques by not requiring crystallization and allowing study of molecules in their natural state. In cryo-EM, a sample is frozen at liquid nitrogen temperatures to vitrify it before being imaged with an electron microscope. This technique has helped determine structures of important proteins and revealed new details about their function. Recent advances in direct electron detectors, image processing, and specimen preparation have led to remarkable improvements in resolution with cryo-EM.
DARK FIELD MICROSCOPY
DARK FIELD MICROSCOPY by SIVASANGARI SHANMUGAM
Dark-field microscopy is ideally used to illuminate unstained samples causing them to appear brightly lit against a dark background.
This type of microscope contains a special condenser that scatters light and causes it to reflect off the specimen at an angle
Microscopy
The document discusses different types of microscopes, including compound microscopes and stereomicroscopes. It describes the key parts and principles of operation. Compound microscopes use multiple lenses to magnify specimens and provide a two-dimensional image. Stereomicroscopes use two optical paths to provide a three-dimensional view of surface details. Examples of uses include biology studies, forensics, manufacturing quality control, and more. The document also discusses who may have invented the compound microscope and provides references for further reading.
Polarising microscope
The document describes the parts and working of a polarizing microscope. It has optical components like polarizers, analyzers and lenses, and mechanical components like the rotating stage. Light from the specimen is polarized and its interaction with the optical components is used to identify properties of minerals and rocks. The polarized microscope allows examination of anisotropic materials and determination of their optical characteristics, which has applications in geology and mineral exploration.
Fluorescence Microscopy
Fluorescence microscopy uses fluorescence to visualize specimens. It works by exciting fluorescent molecules in the sample with high intensity light, causing them to emit light of a longer wavelength. This emitted light is then filtered and used to produce a magnified image of the sample. Modern fluorescence microscopes allow multiple fluorescence filters to be used, and fluorescent markers like dyes, proteins, and antibodies can be introduced to tag specific structures in cells or proteins of interest. This technique is widely used in medical and biological research to study structures and track molecules within living cells.
Introduction to microscopy
This document provides an introduction to microscopy. It discusses different types of microscopes including compound microscopes, which have three main systems - the support system, illumination system, and magnification system. Binocular microscopes have two eyepieces. Important microscope terms are also defined, such as magnification, working distance, resolving power, and numerical aperture. Phase-contrast microscopy is described as a technique that converts phase shifts in light to brightness changes, allowing otherwise transparent specimens to be visible.
Light microscope
LIGHT MICROSCOPY by SIVASANGARI SHANMUGAM
The optical microscope, The functions of a light microscope is based on its ability to focus a beam of light through, which is very small and transparent, to produce an image.
Dark field microscopy
Dark field microscopy is a technique that uses a dark field condenser containing an opaque disk to block light entering the objective lens directly. Only light reflected off the specimen enters the lens, causing specimens to appear bright against a dark background. This technique is useful for viewing unstained, transparent specimens like bacteria, algae, and fibers. It has advantages like viewing details on surfaces but disadvantages like image degradation from non-uniform specimens or particles on the optics. Dark field microscopy finds applications in diagnosing syphilis and viewing various microorganisms, minerals, and cells.
Compound microscope
- The document discusses the compound microscope, its history, parts, functions, and use.
- A compound microscope has more than one lens and was invented in the 1590s in the Netherlands. It allows higher magnification than simple microscopes.
- The main parts are the mechanical stage, optical system with objectives and eyepieces, illumination system, and adjustment controls. Objectives magnify the specimen while eyepieces provide a final view.
- Focusing involves using coarse and fine adjustments to bring the specimen into view at different magnifications including with immersion oil.
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invented in XVII century, and has been improved over the years, resulting in the powerful
modern light microscopes. As individual cellular structures are too small to be seen by the
human eye, microscopy techniques have played a key role in the development of
histological techniques.
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Microscopy
The microscope has evolved a lot from the time of Leeuwenhoek. This presentation gives a brief overview about the types of microscope their principle of function and application.
Bright field microscope and its working principle
The document discusses the bright field microscope. It notes that the bright field microscope is a simple microscope to use, with few adjustments needed. It can view some specimens without staining and does not alter specimen color. However, it has limitations such as distortion with high contrast and inability to view living bacteria. The document describes how bright field microscopy works by using a light source to illuminate the specimen from below, appearing dark against a bright background, and provides examples of specimens that can be observed with it.
Electron microscope
The document discusses the history and development of electron microscopes. It describes how J.J. Thomson discovered electrons in 1897 and how subsequent scientists like de Broglie, Ruska, and Knoll contributed to the development of the first electron microscope in the 1930s. It then explains the basic workings and components of transmission electron microscopes and scanning electron microscopes, how they produce images, and some examples of their applications in biology and materials science.
Light Microscopy
The document provides information about using a compound light microscope, including:
1) The main parts of a compound light microscope are the ocular lenses, body tube, objective lenses, stage, condenser, iris diaphragm, coarse and fine adjustment knobs, and light source.
2) To examine a specimen, the user selects the appropriate objective lens, focuses using the coarse and fine adjustment knobs, and adjusts the condenser and iris diaphragm to control illumination.
3) For oil immersion objectives, oil is added to the slide and specimen to allow for higher magnification imaging.
Microscope ppt, by jitendra kumar pandey,medical micro,2nd yr, mgm medical co...
The document summarizes different types of microscopes used to study microorganisms. It discusses light microscopes like brightfield, darkfield and phase contrast microscopes. It also describes electron microscopes like transmission electron microscopes (TEM) and scanning electron microscopes (SEM) that use electron beams instead of light. TEM images internal structures by transmitting electrons through thin samples while SEM scans sample surfaces using secondary electrons. Sample preparation methods for both light and electron microscopy are also outlined.
Microscope and its types
This document provides information about different types of microscopes. It discusses simple microscopes, which use a single lens for magnification, and their basic parts and working principle. Compound microscopes are then introduced as using multiple lenses to achieve higher magnification than is possible with a simple lens alone. The key parts of a compound microscope are described, including the eyepiece, objective lenses, body tube, arm, and focus adjustments. It explains that light from the illuminator passes through these parts to produce a magnified image for viewing.
Different types of microscopes
This document provides information on different types of microscopy techniques including bright field, dark field, phase contrast, and polarized light microscopy. It begins with explaining the basics of light and microscopy. It then describes each technique in more detail, including their principles, applications, advantages, and how they are set up optically. Bright field microscopy uses illumination and forms a dark image on a bright background. Dark field uses oblique illumination to see small particles as bright objects on a dark background. Phase contrast converts phase differences into contrast changes to see transparent specimens. Polarized light microscopy uses polarized filters to reveal structural details not otherwise seen.
Electron microscope (TEM & SEM)
TEM and SEM are two different Electron microscope used to view objects which otherwise are impossible to see under normal light/ compound microscope.
Dark field microscope
Dark field microscopy produces bright images of unstained samples against a dark background. It works by using a condenser with an opaque disk to block light entering the objective lens directly, allowing only light reflected off the sample to pass through. This causes specimens to appear bright on a dark background. It is useful for viewing transparent or unstained samples like bacteria, cells, and minerals due to the contrast it provides.
Light phase contrast and fluorescence microscopy
Phase contrast microscopy is a type of light microscopy technique that produces high contrast images of transparent specimens without staining. It works by interfering surrounding light waves and sample-diffracted waves to create an image with the inner regions of the sample appearing darker than the outer regions. Fluorescence microscopy uses fluorescent dyes excited by high-energy light to emit lower-energy light, allowing specific structures within samples to be imaged. It is used to study living cells and view genetic material.
1. MICROSCOPY - introduction + principle (Basics)
Basics only
Microscopy is the technical field that uses microscopes to observe samples which are
not in the resolution range of the normal-unaided eye.
Microscope is a scientific-instrument consisting of magnifying lens that enables an
observer to view the minute features distinctly.
In greek, micro = small
skopein = to view.
Recent advancement in microscopic techniques cryoem and super
Cryo-electron microscopy is an emerging technique that uses electron microscopes to image biomolecules such as proteins that have been rapidly frozen to preserve their structure. It overcomes limitations of other techniques by not requiring crystallization and allowing study of molecules in their natural state. In cryo-EM, a sample is frozen at liquid nitrogen temperatures to vitrify it before being imaged with an electron microscope. This technique has helped determine structures of important proteins and revealed new details about their function. Recent advances in direct electron detectors, image processing, and specimen preparation have led to remarkable improvements in resolution with cryo-EM.
DARK FIELD MICROSCOPY
DARK FIELD MICROSCOPY by SIVASANGARI SHANMUGAM
Dark-field microscopy is ideally used to illuminate unstained samples causing them to appear brightly lit against a dark background.
This type of microscope contains a special condenser that scatters light and causes it to reflect off the specimen at an angle
Microscopy
The document discusses different types of microscopes, including compound microscopes and stereomicroscopes. It describes the key parts and principles of operation. Compound microscopes use multiple lenses to magnify specimens and provide a two-dimensional image. Stereomicroscopes use two optical paths to provide a three-dimensional view of surface details. Examples of uses include biology studies, forensics, manufacturing quality control, and more. The document also discusses who may have invented the compound microscope and provides references for further reading.
Polarising microscope
The document describes the parts and working of a polarizing microscope. It has optical components like polarizers, analyzers and lenses, and mechanical components like the rotating stage. Light from the specimen is polarized and its interaction with the optical components is used to identify properties of minerals and rocks. The polarized microscope allows examination of anisotropic materials and determination of their optical characteristics, which has applications in geology and mineral exploration.
Fluorescence Microscopy
Fluorescence microscopy uses fluorescence to visualize specimens. It works by exciting fluorescent molecules in the sample with high intensity light, causing them to emit light of a longer wavelength. This emitted light is then filtered and used to produce a magnified image of the sample. Modern fluorescence microscopes allow multiple fluorescence filters to be used, and fluorescent markers like dyes, proteins, and antibodies can be introduced to tag specific structures in cells or proteins of interest. This technique is widely used in medical and biological research to study structures and track molecules within living cells.
Introduction to microscopy
This document provides an introduction to microscopy. It discusses different types of microscopes including compound microscopes, which have three main systems - the support system, illumination system, and magnification system. Binocular microscopes have two eyepieces. Important microscope terms are also defined, such as magnification, working distance, resolving power, and numerical aperture. Phase-contrast microscopy is described as a technique that converts phase shifts in light to brightness changes, allowing otherwise transparent specimens to be visible.
Light microscope
LIGHT MICROSCOPY by SIVASANGARI SHANMUGAM
The optical microscope, The functions of a light microscope is based on its ability to focus a beam of light through, which is very small and transparent, to produce an image.
Dark field microscopy
Dark field microscopy is a technique that uses a dark field condenser containing an opaque disk to block light entering the objective lens directly. Only light reflected off the specimen enters the lens, causing specimens to appear bright against a dark background. This technique is useful for viewing unstained, transparent specimens like bacteria, algae, and fibers. It has advantages like viewing details on surfaces but disadvantages like image degradation from non-uniform specimens or particles on the optics. Dark field microscopy finds applications in diagnosing syphilis and viewing various microorganisms, minerals, and cells.
Compound microscope
- The document discusses the compound microscope, its history, parts, functions, and use.
- A compound microscope has more than one lens and was invented in the 1590s in the Netherlands. It allows higher magnification than simple microscopes.
- The main parts are the mechanical stage, optical system with objectives and eyepieces, illumination system, and adjustment controls. Objectives magnify the specimen while eyepieces provide a final view.
- Focusing involves using coarse and fine adjustments to bring the specimen into view at different magnifications including with immersion oil.
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invented in XVII century, and has been improved over the years, resulting in the powerful
modern light microscopes. As individual cellular structures are too small to be seen by the
human eye, microscopy techniques have played a key role in the development of
histological techniques.
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A simple microscope uses a single lens to magnify objects and forms a virtual image. It provides low magnification and is used to examine things like skin, algae, and soil samples.
A compound microscope has two lens systems that provide higher magnification by compounding the magnification of the objective and eyepiece lenses. It allows detailed examination of stained slides and is commonly used in biology labs and medical diagnostics.
A stereo microscope uses two separate optical paths and lens systems to provide a three-dimensional view of surfaces. It has lower magnification than compound microscopes and is used for examining things like insects and circuit boards.
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Microscope
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Microscopy - Magnification, Resolving power, Principles, Types and Applications
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Histological tools
This document discusses various histological tools used to study tissues at the microscopic level. It describes light microscopes, which use visible light and magnification to examine thin tissue slices stained with histological dyes. Electron microscopes are also covered, using electron beams instead of light for higher resolution imaging of cell structures. Specific techniques covered include fluorescence microscopy using fluorescent dyes, polarizing microscopy examining birefringence, and transmission electron microscopy producing 2D images of cell organelles. The history and development of microscopy from early simple microscopes to modern compound and electron microscopes is summarized.
Dark field microscope
Darkfield microscopy uses oblique illumination to make specimens appear bright against a dark background. Light is directed around the specimen so that it is scattered and refracted off of it. This allows thin structures like bacteria to be seen more easily compared to brightfield microscopy. Some applications of darkfield microscopy include viewing unstained live samples, motile organisms, fibers, and external surface details of cells. It has advantages like simplicity, quality images, and lack of artifacts, though light levels are lower.
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This document lists the members of Group One for a microscopy course. It includes 9 students' names and registration numbers. The document then provides an overview of different types of microscopes, including light microscopes, electron microscopes, fluorescence microscopes, and dark field microscopes. It describes the basic setup, working principles, uses and comparisons of these microscopic techniques.
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Microscopes and microscopy are introduced. There are two main types of microscopes - light microscopes, which use optical lenses and light, and electron microscopes, which use a beam of electrons. Light microscopes can use different techniques like brightfield, darkfield, fluorescence, and phase contrast. Electron microscopes have higher resolving power and include transmission electron microscopes and scanning electron microscopes. Sample preparation and staining are important for microscopy as they allow small and transparent specimens to be visualized.
Microscopy
Eye microscopy and electron microscopy include differentiation and reflection. Retraction of magnetic fields/electron beams that interact with the image. As well as the scattering of scattered rays or other signals to create the image.
This procedure can be done by inserting a wide-field light sample or by scanning a fine beam over the sample. A microscopy scan probe involves. The interaction of the scanning probe with the surface of the object of interest.
Advances in microscopy transformed living things and exposed the field of histology. And so remain an important strategy for health and natural science.
X-ray microscopy is three-dimensional and unobtrusive. Allowing for repeated photographing of the same sample in situ or 4D subjects. And provides the ability to "see". The sample is readable before devoting it to advanced correction techniques.
The 3D X-ray microscope uses a computed tomography technique, rotating the sample. By 360 degrees and reconstructing images. CT is usually done with a flat panel display. The 3D X-ray microscope uses a series of objectives, e.g., from 4X to 40X, and can include a flat panel.
History of Microscopy
The field of the microscope dates back to at least the 17th century. Early mirrors, single-lens magnifying glasses with limited size. Back to the widespread use of eyeglasses in the 13th century. But the most advanced microscopes first appeared in Europe around 1620 Early.
Microscope doctors included Galileo Galilei, who was discovered in 1610. That he could turn off his telescope to see small objects nearby. And Cornelis Drebbel. Who may have invented the compact microscope in about 1620?
Antonie van Leeuwenhoek developed a simple magnifying microscope. In the 1670s and is often regarded as the first acclaimed microscopist and microbiologist.
Microscope Uses
to view bacteria, parasites, and a variety of human/animal cells
cellular process, cell division
DNA replication
tissue analysis
examining forensic evidence
studying the role of a protein within a cell
studying atomic structures
And in what way are bacteria able to infect human cells, then we use a microscope to study them all. Those studies are done at the micro-level.
We use a microscope to perform the kind of study that we cannot see with the naked eye.
Microscope component
Light
Lence
Optical/Light Microscopy
Bright Field Microscopy
Dark Field Microscopy
Confocal Microscopy
Phase Contrast Microscopy
Fluorescence Microscopy
Electron microscopy
Transmission Electron Microscopy
Scanning Electron Microscopy
Scanning Probe Microscopy
The resolving power of a microscope means
Microscopy - Study
Microscopy is the technical field of using microscopes to view objects that cannot be seen with the naked eye. There are three main types of microscopy - light microscopy, which uses visible light; electron microscopy, which uses electrons; and scanning probe microscopy, which uses a physical probe. Light microscopes like brightfield, darkfield, phase contrast, and fluorescence microscopes are commonly used to view living and stained specimens. Electron microscopes have much higher resolving power than light microscopes and are able to view much smaller structures. Transmission electron microscopes form images using electrons transmitted through thin specimens while scanning electron microscopes form images from electrons emitted from surfaces.
Microscopy - Study
Microscopy is the technical field of using microscopes to view objects that cannot be seen with the naked eye. There are three main types of microscopy - light microscopy, which uses visible light; electron microscopy, which uses electrons; and scanning probe microscopy, which uses a physical probe. Light microscopes like brightfield, darkfield, phase contrast, and fluorescence microscopes are commonly used to view living and stained specimens. Electron microscopes have much higher resolving power than light microscopes and are able to view much smaller structures. Transmission electron microscopes form images using electrons transmitted through thin specimens while scanning electron microscopes form images from electrons emitted from surfaces.
Phase Contrast Microscopy - Microbiology 1st
Phase contrast microscopy uses differences in phase shifts of light waves passing through a specimen to visualize unstained living cells. It allows biologists to study living cells and cell division. Dark field microscopy produces a dark background and bright specimen image using oblique illumination. It is used to view unstained or little absorbed objects like bacteria, algae, and diatoms. Electron microscopy uses a beam of accelerated electrons instead of light for higher resolution imaging of nano-scale structures. Types include analytical electron microscopy, scanning transmission electron microscopy, scanning electron microscopy, and transmission electron microscopy.
2. Microscopy (Biochemistry)
This document provides an overview of microscopy including:
1. It outlines the historical development of the microscope from the 1500s to present.
2. It describes key microscope components and variables like magnification, resolution, numerical aperture, aberration, and contrast.
3. It explains different microscope types like compound light, darkfield, phase contrast, fluorescence, electron, confocal, and scanning probe microscopes as well as their principles and uses.
4. It provides guidance on microscope care and proper storage, handling of lenses, and care of oil emersion objectives and lamps.
Confocal microscopy by prabhat
Confocal microscopy was invented by Marvin Minsky in 1957 and aims to improve resolution over traditional microscopy. It uses point illumination and a pinhole to exclude out-of-focus light and produce thin optical sections and high-contrast images. The key components are a laser light source, dichromatic mirror, pinholes, and photodetector. Confocal microscopy finds applications in cell biology and materials science by allowing optical sectioning and 3D reconstruction. It provides advantages like non-invasiveness, live cell imaging, and depth analysis, but has disadvantages such as photobleaching and loss of intensity.
MICROSCOPY.pptx
The document defines various types of microscopes and microscopy terms. It describes light microscopes like brightfield, phase contrast, and fluorescence microscopes which use lenses and visible light to magnify small specimens. Electron microscopes like transmission electron microscopes and scanning electron microscopes are also covered, which use electron beams instead of light to achieve higher magnifications. Key microscopy terms defined include magnification, resolution, numerical aperture, refractive index, and aberration. Specific uses and working principles of each microscope type are provided.
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Introduction to objective, scope and outcome the subject
Chapter 2
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Introducation to Microscopy
- 1. MICROSCOPY :- PRINCIPLE AND APPLICATION OF LIGHT, PHASE CONTRAST, FLUORESCENCE. -By Narottam V. Priyadarshi
- 2. MICROSCOPY Microscopy is the technical field of using microscopes to view objects and areas of objects that cannot be seen with the naked eye. “Micro” refers to “tiny” and “Scope” refers to “view or look at”. Microscopes are instruments designed to produce magnified visual or photographic images of objects too small to be seen with the naked eye and so they can be studied.
- 3. CLASSIFICATION OF MICROSCOPE Depending on number of lenses i. Single microscope ii. Compound microscope Depending on number of eyepiece i. Monocular microscope ii. Binocular microscope Depending on source i. Light or optical microscope ii. Electron microscope
- 4. TYPES OF MICROSCOPE DEPENDING ON SOURCE Light microscope 1. Bright field Microscope 2. Dark field Microscope 3. Phase contrast Microscope 4. Fluorescence Microscope Electron microscope 1. Scanning tunneling microscope 2. Transmission electron microscope
- 5. LIGHT MICROSCOPY Optical or light microscopy involves passing visible light transmitted through or reflected from the sample through a single or multiple lenses to allow a magnified view of the sample. The resulting image can be detected directly by the eye and/or imaged on a photographic plate or captured digitally.
- 7. BRIGHT-FIELD Bright-field microscopy is the simplest microscope of all the types of optical microscopes. Sample is illuminated using white light from bottom side as shown in Figure and the image is observed from above. The contrast in the specimen is caused by absorbance of some of the transmitted light in dense areas of the sample. The typical appearance of a bright-field microscopy image is a dark sample on a bright background and hence the name.
- 8. APPLICATION OF BRIGHT-FIELD Bright field microscopy is very simple to use with fewer adjustments needed to be made to view specimens. Some specimens can be viewed without staining and the optics used in the bright field technique don’t alter the color of the specimen. It is adaptable with new technology and optional pieces of equipment can be implemented with bright field illumination to give versatility in the tasks it can perform.
- 9. DARK FIELD Microscope image under dark field imaging condition
- 10. DARK FIELD Figure shows the light path in a dark-field microscope. This mode is best for observing the pale objects. Only light rays scattered by specimen enter objective lens and the specimen appears light against dark background. The contrast increases, which enables observation of more details as shown in the Figure , which are not seen in the bright background. This mode is used to observe living, unstained preparations.
- 11. APPLICATION OF DARK-FIELD A dark field microscope is ideal for viewing objects that are unstained, transparent and absorb little or no light. You can also use dark field in the research of live bacterium, as well as mounted cells and tissues. It is more useful in examining external details, such as outlines, edges, grain boundaries and surface defects than internal structure.
- 12. PHASE CONTRAST (a) bright-field and (b) phase contrast microscopes.
- 13. PHASE CONTRAST Phase contrast microscopy is an optical microscopy technique that converts phase shifts in light passing through a transparent specimen to brightness changes in the image. Phase shifts are invisible, but become visible when shown as brightness variations. The basic principle to make phase changes visible in phase contrast microscopy is to separate the illuminating background light from the specimen scattered light (see Figure). This makes up the foreground details, and to manipulate these differently.
- 14. PHASE CONTRAST Light rays in phase produce brighter images, while light rays out of phase produce darker images. Contrast is created because light waves are out of phase. Enhances the contrast between intracellular structures having slight differences in the refractive index. Used to examine living organisms or specimens that would be damaged/altered by attaching them to slides or staining. Excellent way to observe living cells.
- 15. APPLICATION OF PHASE CONTRAST Small unstained specimens such as a living cell can be seen. It makes Highly Transparent objects more visible. Examining Intracellular components of living cells at relatively high resolution. example:- The dynamic motility of Mitochondria, mitotic chromosomes & vacuoles. It made it possible for Biologists to study living cells and how they proliferate through cell division.
- 17. FLUORESCENCE MICROSCOPE Specimen is illuminated with light of a specific wavelength. Light is absorbed by the fluorophores causing an emission of longer wavelengths. The illumination light is separated from weaker emitted fluorescence through the use of a spectral emission filter.
- 18. APPLICATION OF FLUORESCENCE MICROSCOPE Imaging structural components of small specimens such as cells Conducting viability studies on cell populations (are they alive or dead) Imaging the genetic material within a cell (DNA and RNA). To differentiate different types of cells
- 19. THANK YOU…