The Principles and Limitations of Transmission and Scanning Electron Microscopes

Concept 6.1 Biologists use microscopes and the tools of biochemistry to study cells 1. The study of cells has been limited by their small size, and so they were not seen and described until 1665, when Robert Hooke first looked at dead cells from an oak tree. His contemporary, Anton van Leeuwenhoek, crafted lenses and with the improvements in optical aids, a new world was opened. Magnification and resolving power limit what can be seen. Explain the difference. Magnification is the ratio of an object’s image size to its real size. Resolution is a measure of the clarity of the image; it is the minimum distance two points can be separated and still be distinguished

2) Electron Microscope (EM) uses a beam of electrons to resolve electrons, better resolving powers than light microscope M:100,000x RP 0,2 nanometer

Looking through a light microscope at a cell undergoing division, you see that the condensed

The illuminating parts of a microscope enable us to see the detail of the subject placed under the microscope. The three main parts that enable us to do this are: the condenser which illuminates the object that is placed under the microscope, the objectives which forms the magnified image, and the eyepiece which enables us to see the magnified

To make the specimen compatible with both forms of advanced microscopy, they sufficiently prepared samples by coupling the specimen with a fluorescence that was also conductive. This technique was accomplished with the FlouroNanogold label, which contains gold nanoparticles covalently bonded to a fluorescence label. That way, the LM worked as well as the EM for the same set of kinetochores that were being studied. The Hec1 protein was stained in this case because this protein naturally delineates the structures to be studied.

A light microscope was set up with the light on a low setting; one large Daphnia was selected and placed in the centre of a cavity slide by using a pipette.

In electron microscopy, on the other hand, a beam of electrons with a negative charge, instead of light is sent through a very thin slice of the specimen.  Because the electron beam has a far smaller wavelength than light used in light microscopy, it achieves far better resolution, “the current resolution of limit of the best electron microscope is approximately 0.05 nm atomic resolution, and 4000X better magnification than that of a conventional light microscope”(BSP, S., 2010).  This means that with an electron microscope you can potentially achieve enough magnification to observe the shape of the protein machinery that carries out the work inside of cells. The major limitation of electron microscopy is that specimen

By using their microscopes, they found that every living plant and animal they examined was made of cells. As microscopes were improved, scientists were able to see smaller and smaller organisms. They found that no matter how large or small the organism was, it was made of cells, leading to cell theory. For example, a German biologist, Theodor Schwann discovered that all plant and animal cells were divided into cells by looking through his microscope. He also discovered that the cell is the basic unit of organization in organisms. Cells can be grouped together to form tissues, which can in turn be grouped together to make an organ. Organs can be grouped together to form a system, which is part of an organism. He was able to use microscopes to see the ways that cells work and help to determine which kind of microorganisms (bacteria) is causing the disease and making people ill. This is particularly valuable in the study of the components of organisms, where physicians are able to overcome a treatment of method to kill disease cells and restore people¡¦s health. The microscope revealed not only the cellular structure of human tissues, but also the organisms that cause diseases. The discovery of cells led scientists to study cells and discover more information about cells; this, allowed scientists to find ways to prevent or cure diseases. The use of microscopes has made many

A small square of a red onion skin (membrane) was observed under a microscope at high power (X40) magnification. The observation showed a large number of onion cells. The structure of one onion cell had a general rectangular shape with a developed cell wall, which gives the rectangular shape to the cell and a cell membrane just beneath it.

“The short (about 1 millimeter) focal lengths of the lenses would have necessitated placing the eye almost in contact with the lens” ("Anton van Leeuwenhoek"). Leeuwenhoek obtained the clear image by carefully moving the angle of lighting left and right. Leeuwenhoek's techniques of lighting samples under the microscope are still not well known today. This was the only secret that he took to his grave. Even though the simple microscope was difficult to use, scholars visited Leeuwenhoek to be educated on his design. Leeuwenhoek went all over the world giving demonstrations about his microscope for high-ranking people. Without Leeuwenhoek’s simple microscope, microbiology today would not be as advanced.

Resolution is related to the numerical aperture of the objective lens – the higher the numerical aperture, the better the resolution; and the wavelength of light passing through the lens – the shorter the wavelength, the better the resolution.

But first, let us talk about the discovery of cells and the cell theory. Robert Hooke, an english scientist, was the man who first discovered the cell in 1665, proof being a book he released at that time called Micrographia. In this book, Hooke gave 60 observations of random objects under a compound microscope with a magnification of 30x. Because of this, he was not able to see the internal structures in the cell, like nuclei and vacuoles, and what he proclaimed to be cells were just empty cell walls of plant tissues. He shared his observations with The English Royal Society, until they started receiving letters from a scientist named Anton van Leeuwenhoek. The letters stated that Anton made use of a microscope containing improved lenses that magnified objects up to 275x, enough to identify the living parts of a cell. He kept on sending

NOTE: Answer Question A only if you used a compound light microscope for this experiment.

Most microscopes, including those in schools and laboratories today, are optical microscopes. They use glass lenses to enlarge, or magnify, an image. An optical microscope cannot produce an image of an object smaller than the length of the light wave in use. To see anything smaller than 2,000 angstroms (about 1/250,000 of an inch) a wave of shorter length would

“The prefix ‘nano’ stems from the ancient Greek word for ‘dwarf’. In science, it means one billionth (10 to the minus 9) of something, thus a nanometer (nm) is one billionth of a meter, or 0.000000001 meters. A nanometer is about three to five atoms wide, or some 40,000 times smaller than the thickness of human hair. A virus is typically 100 nm in size.” (Paddock) “The ability to manipulate structures and properties at the nanoscale in medicine is like having a sub-microscopic lab bench on which you can handle cell components, viruses or pieces of DNA, using a range of tiny tools, robots and tubes.” (Paddock) There is one type of microscope in the world that has the ability to see things at the nano scale. That microscope is a scanning tunneling microscope. It has the ability to zoom in on an object by 1,000,000 times as the average high school and college microscope only reaches 100(Nano.gov).