Introduction:
In order to study the cell and its component, it has to be visualised and displayed in details. In this practical class, we will be looking at different microscopic techniques that visualise the cell structures and identify its features. As most cells are very small, they cannot be seen with naked eyes and therefore need to be magnified. Light microscopy was first used to magnify the image of the cells using stains. However, some tissue and subcellular structures are too small to be seen even under the light microscope. Therefore another technique was found to visualise the cell in more details. To study the smaller features of the cell, electron microscopy are used. Electron microscopy use electron beam to visualise the specimen. Electron microscopy can only magnify thin structures, therefore fluorescent microscopy are used to visualise the thicker structures. Fluorescent microscopy visualise the structures that emit light by allowing the light to get through the specimen.
Methods
Question 1:
Briefly describe how immune-staining with an antibody is done.
Immune-staining with an antibody is done by binding the antibody to a specific antigen. This is carried out by adding a staining fluorescent dye to a thin section of a tissue or cell. A secondary antibody is coupled to a fluorescent marker (1) that can attached to the primary antibody and increase the light signal. Hence, increase visibility.
Question 2: Fluorescence/ Immunofluorescence Complete figure 1 below (ie draw in the light paths).
Figure 1. Diagram showing the light path for both A. Transmitted Light Illumination and B. Fluorescent Illumination (Epi-illumination) in an inverted microscope. Lines in red indicate the path for light reaching t...
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...its functions.
References:
(1) Dr. Potonik S, Lecture 2 Notes, BIOL2319 “Visualising cells”, 4 March 2014
(2) http://orthomolecular.org/nutrients/glycogen.html
(3) http://www.plosone.org/article/info%3Adoi%2F10.1371%2Fjournal.pone.0077166
(4) http://www.princeton.edu/~achaney/tmve/wiki100k/docs/Barr_body.html
(5) http://cellmontage2.cira.kyoto-u.ac.jp/cgi-bin/cellinf.cgi?cellname=basophil
(6) http://www.google.com.au/url?sa=t&rct=j&q=&esrc=s&frm=1&source=web&cd=3&ved=0CD8QFjAC&url=http%3A%2F%2Fwww.ocr.org.uk%2FImages%2F140893-unit-r074-calculating-magnification-lesson-element-teacher-instructions.pdf&ei=wwUpU63QGMqQlQXn8oGgBg&usg=AFQjCNGRnI5IFykp3Qso-eihUA0_x7gl_w
(7) https://www.jic.ac.uk/microscopy/intro_LM.html
(8) http://www.ammrf.org.au/myscope/sem/background/
(9) http://en.wikipedia.org/wiki/High-resolution_transmission_electron_microscopy
In the pages of “A Conspiracy of Cells” that were available to us, there were several differences in both writing style and content, compared to Skloot’s, “The Immortal Life of Henrietta Lacks.” One of the biggest differences is in Michael Gold’s, “A Conspiracy of Cells,” he focuses more on the Hela cells and the research, while in Skloot’s “The Immortal Life of Henrietta Lacks” Skloot focuses more on the Lacks family.
Cells are the basic units of life, and they can be found everywhere that you look and go. Most cells cannot be viewed without the aid of a microscope. Plant and animal cells are very different not only in their structure shape but in their functions as well. The diagrams found in the book on pages 65-66 are described as generalized cells that are used for study purposes (Mader & Windelspecht, 2016).
As it became my hobby to study quite a few microscopic and gross preparations for hours every day. Working under a fine supervision of my pathology professor Dr. Bekhtereva, made me aware of my ability to identify and follow a specific pattern in a slide. My mentor emphasized how important it is to be able to combine this innate visual ability with rigorous scientific
Immunohistochemistry is a technique that involves the use of antibody-antigen interactions in order to identify cellular and tissue constituents. One way this can be done by labelling known antibodies with enzymes which produce a coloured product after reacting and then monitoring the sample to see whether a reaction has taken place. In immunohistochemistry, the preservation of the antigenic determinants, also known as epitopes, and binding sites is vital in order to ensure that an accurate result is obtained. However, tissue samples need to go through various processes in a pathology laboratory before they are ready for testing and some of these processes can alter the structure of the epitopes so it’s important to consider this when choosing reagents in order to reduce the risk of this happening.
... produces can be measured. (Chesney and Folkman, 1999) A positive result is identified by a polymerase chain reaction and the presence of the specific antibody.
One can almost feel the searing penetration of Lewis Thomas’ analytical eye as it descends the narrow barrel of the microscope and explodes onto a scene of vigorous, animated, interactive little cells—cells inescapably engrossed in relaying messages to one another with every bump and bounce; with every brush of the elbow, lick of the stamp, and click of the mouse…
"Method of the Year 2010: Optogenetics - by Nature Video." YouTube. Macmillan Publishers Ltd, 17 Dec. 2010. Web. 12 Apr. 2014. .
Immunosensors make use of antigen-antibody interactions [1, 2] to detect a wide range of analytes which are of great interest in medical diagnostics, environmental analysis, and forensic medicine [3, 4], including pathogens [5], drugs [6], bacteria [7], toxins [8], and biomarkers [9]. Overall, immunosensors employ the same chemical approach of earliest immunoassays, but offer quicker and simpler analytical procedures that may be conducted at the point-of-care [10]. Immunosensors use an antibody immobilized as a transducer. Antigen-antibody binding event result in electrical or optical changes.
to construct and or maintain the cell membrane. In a microscopic view of the cell membrane we can
For the production of the monoclonal antibodies, B-cells are removed from the spleen of an animal that has been challenged with the relevant antigen. These B-cell are then fused with myeloma tumor cells that can grow indefinitely in cultures (myeloma is B-cell cancer). This fusion is performed by making the cell membranes more permeable. The fused hybrid cells, being ous cancer oils cells, will multiply rapidly, as a result, large amounts of the desired antibodies will be
Many stains and dyes were used in the experiments. They were water, methylene blue, salts, and iodine. In our studies of cells, we conducted three experiments to test the different features of cells. The first two experiments were on how membranes were selectively permeable, diffusion, and osmosis.
The Animal Cell is a little bit different than the Plant Cell for only a couple of reasons. One is how the Plant Cell has a cell wall and the Animal Cell doesn’t. The cell wall protects and gives structure to the cell. Then there is the Nucleus, which serves as a control center for the cell. Inside the Nucleus there are one or more Nucleoli. They are dense, granular bodies that disappear at the beginning of cell division and reappear at the end. Then you have the Cytoplasm. This is the watery material lying within the cell between the cell membrane and the nucleus. The Cytoplasm also contains organelles, which have specific functions in the cell metabolism. Then there are the Golgi Bodies, which serve as processing, packaging, and storage for the cell. These organelles package and ship things out. Another parts of the cell, a very important one in fact, are the Lysosomes. These organelles are used to break things down and contain enzymes.
Image intensification is the process of converting x-ray into visible light. “Early fluoroscopic procedures produced visual images of low intensity, which required the radiologist's eyes to be dark adapted and restricted image recording. In the late 1940s, with the rapid developments in electronics and borrowing the ideas from vacuum tube technology, scientists invented the x-ray image intensifier, which considerably brightened fluoroscopic images” (Wang & Blackburn, 2000, np). We will explore the image-intensification tube, the various gain parameters associated with the tube, and the magnification mode of the image intensifier.
The clusters inside the cells look like pairs of threads wound around each other in a helix. The tangles consist of a protein called tau. Tau binds to another protein called tubulin. Tubulin then forms structures called microtubules which run through cells, giving support and shape. Also the microtubules provide pathways for nutrients and other molecules to travel through.
This report provides an insight into the differences in the structure of cells and the way that they carry out their internal mechanisms. Cells form the basis of all living things and they are the smallest single unit of life. Cell biology is the study of cells and how they function, from the subcellular processes which keep them functioning, to the