Unit 1 How to Use Microscope
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The document is a ratification page for a report on using a microscope. It includes the name, student ID number, class, and group of the student who authored the report, Shally Rahmawaty. It also includes the signatures and IDs of the assistant and assistant coordinator who reviewed the report and accepted it, dated November 2013 in Makassar.
Unit 1 How to Use Microscope
- 1. RATIFICATION PAGE Complete report of Basic Biology with title “How to Use Microscope”, which made by: Name : Shally Rahmawaty Reg. Num (ID) : 1312440001 Class : Physics ICP A Group : II After checked by assistant and assistant coordinator, so this report accepted, Makassar, November 2013 Assistant Coordinator Assistant Adi Putra Rahman Nur Hafri Mulyani ID: 091414021 ID : 1114040204 Known, Lecture of Responsibility Sitti Zaenab, S.Pd, M.Pd ID: 19810302 200912 2 003
- 2. CHAPTER I INTRODUCTION A. Background So many things that can not be seen with naked eye. Examples constituent structure of living things. Bodies of living things (organisms) are composed of cells. The cell is the smallest unit of an organism. Due to the very small size, the cell can not be seen with the eyes directly so it takes a special tool to be able to see it. This has led researchers to look for a tool that can be used to see objects that are very small. Until then found that the microscope can be used to see objects as small as a cell size. Microscope is an instrument designed to make fine details visible. A microscope is an object used to see objects that are too small to be viewed using naked eyes. The purpose of a microscope is to enlarge and make the tiny objects visible and it is mainly used in scientific research. Historically, the study of cell biology could not have happened without the invention of microscopes because cells were not known to exist before Antonin van Leeuwenhoek and Robert Hooke saw them in their primitive microscopes. The early simple “microscopes” which were really only magnifying glasses had one power, usually about 6x – 10x. One thing that was very common and interesting to look at was fleas and other tiny insects. These early magnifiers were hence called “flea glasses”. The “simple microscope” or magnifying glass reached its highest state of perfection, in the 1600’s, in the work of Anton von Leeuwenhoek who was able to see single-celled animals (“animalcules”) and even some larger bacteria. Meanwhile, the cells were first discovered by Robert Hooke in 1655 by using a simple microscope. Robert Hooke saw cork cells the cells of the incision. The microscope consists of several components, namely optical and mechanical components that have different functions. Before performing the
- 3. observations, we must know the function the parts of the microscope to help us in making observations. In addition, we also need to know how to use, how to clean and how to keep a microscope. A. Purpose The objective in this experiment is mean to skillfully use and safely biological microscope to see the simple preparations. B. Benefit The benefits of this practicum is to determine, recognize microscope parts and their functions, and how to use it. And understand the elements contained in the object of research.
- 4. CHAPTER II PREVIEW OF LITERATURE The cell theory: Naturalists and scientists continued observing the living microscopic world using glass lenses. In 1838, German scientist Matthias Schleiden carefully studied plant tissues and concluded that all plants are composed of cells. A year later, another German scientist, Theodor Schwann, reported that animal tissues also consisted of individual cells. Prussian physician Rudolph Virchow proposed in 1855 that all cells are produced from the division of existing cells. The observations and conclusions of these scientists and others are summarized as the cell theory. The cell theory is one of the fundamental ideas of modern biology and includes the following three principles: 1. All living organisms are composed of one or more cells. 2. Cells are the basic unit of structure and organization of all living organisms. 3. Cells arise only from previously existing cells, with cells passing copies of their genetic material on to their daughter cells. (Glencoe, 2008) Cell walls were first seen by Robert Hooke in 1665 as he looked through a microscope at dead cells from the bark of an oak tree. But it took the wonderfully crafted lenses of Antoni van Leeuwenhoek to visualize living cells. Imagine Hooke’s awe when he visited van Leeuwenhoek in 1674 and the world of microorganisms what his host called very little animalcules was revealed to him. (Campbell, 2010) A microscope is an instrument designed to make fine details visible. The microscope must accomplish three tasks: produce a magnified image of the specimen (magnification), separate the details in the image (resolution), and render the details visible to the eye, camera, or other imaging device (contrast). Because of the limited ability of the eye’s lens to change its shape, objects brought very close to the eye cannot have their images brought to focus on the retina. The accepted minimal
- 5. conventional viewing distance is 10 inches or 250 millimeters (25 centimeters). More than five hundred years ago, simple glass magnifiers were developed. These were convex lenses (thicker in the center than the periphery). The specimen or object could be focused by use of the magnifier placed between the object and the eye. These “simple microscopes”, along with the cornea and eye lens, could spread the image on the retina by magnification through increasing the visual angle on the retina. The “simple microscope” or magnifying glass reached its highest state of perfection, in the 1600’s, in the work of Anton von Leeuwenhoek who was able to see single-celled animals (“animalcules”) and even some larger bacteria. The image produced by such a magnifier, held close to the observer’s eye, appears as if it were on the same side of the lens as the object itself. Such an image, seen as if it were ten inches from the eye, is known as a virtual image and cannot be captured on film. These magnifiers had severe limitations in specimen positioning, illumination, lens aberrations, and construction. (Mortimer, 2003) Three important parameters in microscopy are magnification, resolution, and contrast. Magnification is the ratio of an object s image size to its real size. Light microscopes can magnify effectively to about 1,000 times the actual size of the specimen; at greater magnifications, additional details cannot be seen clearly. Resolution is a measure of the clarity of the image; it is the minimum distance two points can be separated and still be distinguished as two points. For example, what appears to the unaided eye as one star in the sky may be resolved as twin stars with a telescope, which has a higher resolving ability than the eye. Similarly, using standard techniques, the light microscope cannot resolve detail ner than about 0.2 micrometer ( m), or 200 nanometers (nm), regardless of the magnification. The third parameter, contrast, accentuates differences in parts of the sample. Improvements in light microscopy have included new methods for enhancing contrast, such as staining or labeling cell components to stand out visually. (Campbell, 2010) Optical microscope consists of two, namely biological microscopes and stereo microscopes. Biological microscope is used for observation of transparent thin objects. Irradiation is given from below with natural light or lights. This biological
- 6. microscope eyepiece and generally has an objective lens with a magnification power as follows : 1. 4x objective with 10x eyepiece, 40x magnification 2. Ocular 10x by objective 10x, 100x magnification 3. Ocular 40x by objective 10x, 400x magnification 4. 100x objective with a 10x eyepiece, magnification 1000x Objective of the most powerful optical microscope objective emersi 1000x called, because of its use must be with emersion oil and how to wear it especially well. Stereo microscope is used for observation of objects that are not too large, transparent or not. The rays can be set from above or from below with natural light or lights. Has two objective and two ocular, in order to obtain a three-dimensional image with the observation of two-piece eye. Strength is not too strong magnification 1x or 2x general objective with 10x eyepiece or 15 times. (Tim penyusun, 2013) In a light microscope (LM), visible light is passed through the specimen and then through glass lenses. The lenses refract (bend) the light in such a way that the image of the specimen is magnified as it is projected into the eye or into a camera. Until recently, the resolution barrier prevented cell biologists from using standard light microscopy to study organelles, the membrane-enclosed structures within eukaryotic cells. To see these structures in any detail required the development of a new instrument. In the 1950s, the electron microscope was introduced to biology. Rather than light, the electron microscope (EM) focuses a beam of electrons through the specimen or onto its surface. Electron microscopes have revealed many organelles and other subcellular structures that were impossible to resolve with the light microscope. But the light microscope offers advantages, especially in studying living cells. A disadvantage of electron microscopy is that the methods used to prepare the specimen kill the cells. (Campbell, 2010) Another type of microscope, the scanning tunneling electron microscope (STM), involves bringing the charged tip of a probe extremely close to the specimen so that the electrons “tunnel” through the small gap between the specimen and the tip. This instrument has enabled scientists to create three-dimensional computer images
- 7. of objects as small as atoms. Unlike TEM and SEM, STM can be used with live specimens. The atomic force microscope (AFM) measures various forces between the tip of a probe and the cell surface. (Glencoe, 2008)
- 8. CHAPTER III OBSERVATION METHOD A. Time and Place Day/date : Friday, November 22nd 2013 Time : 04.00-05.50 PM Place : Green House of Biology, FMIPA UNM B. Tools and Materials 1. Tools provided by laboratory a. Biological microscopes b. Toolbox contains of: 1) Glass object 2) Glass cover 3) The cup petri 4) Tweezers 5) Pipette 2. Tools provided by student a. Razor blade b. Flannel fabric c. Roll tissue d. Drawing book and pencil e. Toothpick 3. Materials provided by laboratory a. Distilled water b. Filter paper or blotting paper c. Cotton 4. Materials provided by student a. Leaf of Allium cepa b. Leaf of Hibiscus rosa-sinensis
- 9. c. Leaf of Sechium edule C. Work Procedure 1. Setting up the microscope 1.1 The microscope is placed on the desk right in front of us. 1.2 Body of microscope cleaned with flannel. Never rub the lens with flannel. 1.3 The tool box is opened, remove the cup containing the stained glass objects and glass cover. Glass objects are cleaned with a cotton cloth or filter paper. 1.4 On the table there are only a microscope work, toolbox with its contents, the handbook and notes, materials for the lab. Other excluded at other places that have been provided. 2. Regulate the entry of light into the Tubus 2.1 The state of lab space considered, where the direction of the brighter light (from the front, left, or right), Mirror microscope light source is directed to. Diaphragm plate is opened or rotated at a medium-size hole position. Microscope condenser which has set its position to approach the preparation and use flat mirrors. To microscope without condensers used a concave mirror. 2.2 The position of the objective lens revolver arranged so that most shortfacing bench preparations until a click. 2.3 Tubus reduced to the distance to the objective end of the preparation table or Tubus down 5-10 mm maximum. 2.4 We telescoped through the eyepiece with the left eye without squinting (need practice) will appear white circular field. 2.5 Prepared microscope used to observe preparations. 3. How to adjust the distance of the lens with the preparation 3.1 With hands, coarse adjustment rotated or macrometer regulator towards masters fingers, Tubus down, distance objective with a smaller preparation table, do the opposite.
- 10. 3.2 Glass objects containing preparations on the table amid hole desk placed, glass objects clamped with clip so as not to shake. 3.3 Should be noted the distance objective with glass objects no more than 10 mm. If the distance was great, then rotated to lower micrometer tubus while viewed from the side closer to the tip of the objective glass objects up to a maximum 5-10 mm. 3.4 We telescoped through the eyepiece while hand turning macrometer by raising tubus slowly. 3.5 Magnification of eyepiece and objective is checked. Calculated the magnification of shadow. 3.6 Removed preparations that have been observed. 4. Making simple preparations Observe the cotton fiber 4.1 The cleaned glass objects were taken and held evenly as possible. 4.2 Spilled glass of clear water or a drop of distilled water in the midst of. 4.3 With tweezers, one crate material removed and placed amid water droplets. 4.4 The hand that helds the cover glass door between the master with the index finger on the opposite side or edge. 4.5 Side of the cover glass on glass objects touched by water droplets with a slope of 450 then released so right cover water droplets. Excess water that seeps on the edge of the glass is absorbed by the filter paper. 4.6 Artificial preparations have been made, fitted on a table and observed preparations as steps 3.2, 3.3, 3.4, and 3.5. 5. Observed enlargement 5.1 Shadow that appears to be raised again. Position preparations or tubus not be touched. 5.2 Nosepiece rotated such that the objective lens longer (stronger) perpendicular to the preparation table until there was a click.
- 11. 5.3 We telescoped while turning the micrometer until the image appears larger. 5.4 At the end of the activity using a microscope, there are some things must be considered: a. Mixture should not be stored on the table preparations, should be excluded. b. Mixture should be cleaned with a wet filter paper or cotton cloth (glass objects + glass cover). Store in a petri dish and put in the gear box. c. Body of microscope is cleaned with a flannel cloth. Lowering tubus low as possible. d. Stored microscope into its box. e. All the equipment has been used, cleaned with cotton cloth and kept in a box. f. Equipment itself, kept themselves to be used in the next activity. g. The rest of the materials are not used anymore discarded rubbish place available.
- 12. CHAPTER IV RESULT AND DISCUSSION A. Result 1. Microscope Observation Picture Note 1. Eyepiece (Ocular lens) 2. Body Tube 3. Coarse adjustmet knob 4. Fine adjustment knob 5. Arm 6. Stage 7. Inclination joint 8. Mirror 9. Base 10. Diaphragm 11. Condensor
- 13. Comparison Picture Note 1. Eyepiece (Ocular) 2. Nosepiece (turret) 3. High power objective 4. Low power objective 5.Diaphragm 6. Light 7. Base 8. Fine adjustment knob 9. Coarse adjustment knob 10. Stage 11. Stage Clips 12. Arm 13. Body Tube
- 14. 1. Observation picture of Allium cepa Observation Picture Note Magnification: 10 x 10 Note Comparison Picture 1. Nucleus 2. Cell wall 3. Liquid (sitoplasm) 2. Observation picture of Hibiscus rosasinensis Observation Picture Magnification: 10 x 10 Comparison Picture Note Note 1. Epidermic
- 15. 3. Observation picture of Sechium edule Observation Picture Note Magnification: 10 x 10 Comparison Picture Note 1. Nodus B. Discussion The microscope is an optical instrument consisting of two convex lenses are used to see very small objects. The eyepiece lens is adjacent to the eye and objective lens adjacent to the object being observed. There are two main parts to the microscope, namely: Optical part, which consists of a condenser, mirror, objective lens and eyepiece. And the non-optical or mechanical, consisting of leg and arm of microscope, the diaphragm, the stage / counter preparations, smooth and rough player, slide clamp, and a light source. In which the function the part of microscope are: Arm - The arm connects the body tube to the base. The user must hold this part in order to move the microscope from one place to another.
- 16. Base - As the name suggests, the base is the lowest portion on which the whole structure of the microscope rests. Eyepiece/Ocular lens - It is through the eyepiece that we look at the specimen placed on the stage of the microscope. It contains two or more lenses. The most common magnification for the eyepiece is of 10x, however, they can also be of 2x and 5x. An eyepiece is a removable part that can be interchanged with another one of a different magnification. Objective Lenses - A standard compound microscope contains two primary objective lenses, which can have magnification of 4x, 5x, 10x, 20x, 40x, 50x, and 100x. The magnification values are written on the side of each lens. The objective turret to which these lenses are attached, can be manually rotated to get the lens to give the desired magnification and focus of the specimen. Stage - is the platform below the objective lens on which the object or specimen to be viewed is placed. There is a hole in the stage through which light beam passes and illuminates the specimen that is to be viewed. Stage Clips - There are two stage clips, one on each side of the stage. Once the slide containing the specimen is placed on the stage, the stage clips are used to hold the slide in place. Diaphragm - is located on the lower surface of the stage. It is used to control the amount of light that reaches the specimen through the hole in the stage. Illuminator - Simple compound microscopes have a mirror that can be moved to adjust the amount of light that is focused on the specimen. However, some advanced types of compound microscopes have their own light source. The Adjustments - There are two adjustment knobs, the fine adjustment knob and the coarse adjustment knob. The coarse adjustment knob helps in improving the focus at a low power, whereas the fine adjustment knob helps in adjusting the focus of the lenses with higher magnification. Mirror: Reflects light upward through the diaphragm Body Tube - It is the part of the microscope that holds the eyepiece.
- 17. Based on practice that has been implemented, has obtained results are described in the form of an image and description. 1. Leaf of Allium Cepa The observations made by using thin slices leaf of Allium Cepa with a magnification of 10 x 10, looks cells shaped like bricks. Cell wall serves to reinforce and give shape to the cell and protects the cell shape. From the magnification could be seen clearly the shape of this cell including its nucleus, its wall cell, and its sitoplasm, its cell is long and combined like set of gypsum. 2. Leaf of Hibiscus rosa-sinensis Observations using Hibiscus rosa-sinensis preparations with a magnification of 10 x 10, it will look tricomhes and cells protector. From the magnification could be seen that its shape is nearly round, there were thin feathers, in the surface there were dots and its important in reproduction process. 3. Leaf of Sechium edule In observation of preserved Sechium edulewith magnification of 10 x 10, which seems just a collection of cells. From the magnification we can see the inside of the pumpkin leaves the bone, such as glands, the cell wall, and itchy hair. We can see trichome or fine hair from the leaves of the pumpkin that is above the cell wall.
- 18. CHAPTER V CONCLUSION AND SUGGESTION A. Conclusion Based on practicum we have done, we can know a microscope is a tool used to see objects that are too small to be viewed using naked eyes. We can recognize microscope parts and their functions. Because with knowing each part, we can skillfully operate the microscope well. B. Suggestion 1. For the assistant, in order to provide directives and guidance regarding the experiments to be performed. And also have to be patient to face the aprentice. If there is an error in the process of conducting experiments, assistant may be able to help and give a solution of the problem. 2. To the apprentice, if at the time assistant to provide guidance, the apprentice must consider with the explanation given. So that, during the experiment was not a mistake. Apprentice must also perform experimental procedures were in accordance with keeping the tools used are not damaged. In addition, the apprentice must also maintain the cleanliness of the laboratory. 3. To the laboratory, should be able to provide tools that will be used by the apprentice to experiment undertaken in order practicum goes well.
- 19. BIBLIOGRAPHY Abramowitz, Mortimer. 2003. Basics and Beyond volume 1. www.micro.magnet.fsu.edu/primer/pdfs/basicandbeyond.pdf, November, 28th 2013. Campbell. 2010. Biology. Jakarta: Erlangga. M. Raith, Michael. 2011. Guide to Thin Section Microscopy. www.minsocam.org/msa/content/.../Thin Section Microscophy.pdf, November, 28th 2013. Science, Glencoe. 2008. Biology. United States of America: National Geographic. Tim penyusun. 2013. Biologi Umum. Makassar: Jurusan Biologi FMIPA UNM. .
- 20. APPENDIX Questioner 1. Write down the name of the optical part microscope! Answer: - Ocular lens - Objective lens - Condenser - Mirrors 2. Write the name of the mechanical parts of microscope! Answer: Base, object tweezers, mechanical drives, makrometer, micrometers, tubus, revolvers, pole, arm microscope, regulator condenser, diaphragm, preparation table, preparation table hole. 3. If the shadow in the terrain view would be shifted to the left - front , glass objects which direction /dosage should slide? Why it can be like that? Answer: Glass items should be in the slider to the right-rear. That's because the shadow is in its nature received by the lens upside down, magnified, and real. 4. Write a negative effect on the lens in the microscope that rub with a cloth or plain paper / rude! Answer: If the lenses in rubbing with a cloth or plain paper/rough, can cause the lens to be damaged or scratched and the observed image would become clear. This can affect the research conducted.