DEVELOPMENT FOUNDATION IN SCIENCE BIOLOGY PRACTICAL 1 (PRACTICAL 2: MICROSCOPY) (FBIP 1112)
NAME: AMBIGAI A/P GANESAN
PRACTICAL TITLE: MICROSCOPY MATRIC NUMBER: U0230002 COHORT: FIS 26 PRACTICAL DATE: 16/8/2023 SUBMISSION DATE: 23/8/2023 LECTURER’S NAME: SIR MOHD SHAHMEN MOHD YAZAM 2.1 OBJECTIVES 1. To identify each of microscope components and their functions. 2. To practice the operating procedure of light microscope. 3. To perform wet mount procedure. 2.2 INTRODUCTION One of the major instruments used in microbiological labs is a low magnification microscope. The examination process will be faster and the degree to which an experiment has been successful will also be determined by proper use of a microscope. The following ways of using the microscope shall be used, such as Always ensure the slide is clean and dry before placing it on the stage. Ensure the lenses are free from dust, after using the oil immersion lens, ensure the lens is free from oil residue before you store it. Always use lens cleaning paper to clean the lens, when handling a microscope, hold the limb with right hand and use the other hand to support from the bottom. It must be carried vertically whenever possible, a microscope needs to be placed about at least 15cm from the edge of a table, do not touch lens with finger and keep a microscope away from acidic substances and cover the microscope with enclosed cover properly for storage. 2.3 MATERIALS 1. Histology slides 2. Glass slide 3. Cover slip 4. Lens cleaning paper 5. Immersion oil 6. 95% alcohol 7. Distilled water 8. Tissue roll 9. Cutter/scissors 10.Disposable dropper 11. Light microscope 2.4 METHODOLOGY ➢ Microscope Component ➢ Uses of Microscopy Material and Apparatus 1. Histology slides 2. Glass slide 3. Cover slip 4. Lens cleaning paper 5. Immersion oil 6. 95% alcohol 7. Distilled water 8. Tissue roll 9. Cutter/scissors 10.Disposable dropper
Method: Wet Mount Procedure 1. A drop of water was placed on the centre of the slide. 2. The specimen is placed into the drop of water and if the specimen floats, another drop of water was dropped on top of it. This reduces the possibilities of air bubbles forming. 3. The cover glass was carefully lowered so that it touches with one side the drop of water. The cover slip should form an angle of about 45 degrees with the slide. the cover glass was touched on the sides only to prevent finger prints. Alternatively, tweezers were used to hold the cover glass. 4. Then, the cover slip was lowered completely. The cover slip was placed at an angle prevents the formation of air-bubbles. 5. Excess water was removed with a filter paper or tissue paper. Microscope Observation Procedure 1. The revolving turret/nosepiece was turned so that the lowest power objective lens (ex. 4x) is clicked into position. 2. The microscope slide was placed on the stage and fasten it with the stage clips. 3. The objective lens and the stage was looked from the side and turn the focus knob so the stage moves upward. The objective was moved up as far as it will go without letting touch the coverslip. 4. The eyepiece/ocular lens was looked through and the focus knob was moved until the image comes into focus. 5. the condenser and light intensity was adjusted for the greatest amount of light. 6. The microscope slide was moved around until the sample is in the centre of the field of view (what you see). 7. The focus knob was used to place the sample into focus and the condenser and light intensity was readjusted for the clearest image (with low power objectives you might need to reduce the light intensity or shut the condenser). 8. The next objective lenses can be changed When you have a clear image of your sample with the lowest power objective. The sample might need to be readjusted into focus and/or the condenser and light intensity readjusted. If you cannot focus on your specimen, steps 3 through 5 should be repeated with the higher power objective lens in place. Do not let the objective lens touch the slide. 9. When finished, the stage was lowered, the low power lens was clicked into position and the slide was removed. 2.5 RESULTS ➢ Diagram of Light Compound Microscope ➢ Drawing of word 2.6 DISCUSSION Microscope is an instrument for forming magnified images with high resolution for detail. The purpose of using the microscope is to observe small things that cannot be observed by naked / unaided eye. There are few precautions in using the microscope. Ensure to turn the voltage down to the lowest before turning off the microscope. Do not turn the power on and off, off and on immediately it will shorten the life span of the bulbs and may cause damage to the electrical system. Ensure that the platform is being moved from up to down forth as this to avoid the slide broken. Should always use low power objective before changing to the high-power objectives. Clean up the microscope objective (100x) right after using it; ensure that no more immersion oil residue left on it. Never leave the microscope open without any covers. When you have finished, cover it with the enclosed dust cover properly. To clean the microscope, we must follow some rules. If dust is found on the optical surface, try to remove it by air blower or compressed air. Remove the fingerprints or grease stain by wiping it with a lens paper or dip a lens paper with a small amount of absolute alcohol and clean the lens surface carefully. From the results that obtained, for the 40X magnification, the word ‘PROFESSIONAL’ can be seen clearly and fully. In 100X magnification, the word ‘PROFESSIONAL’ can be seen half as ‘FESSIO’. While in 400X magnification, only dotted lines can be seen and in 1000X magnification the dotted line can be seen more clearly. 2.7 CONCLUSION In biomedical science courses, as well as in research and testing facilities, the light microscope is an especially useful instrument for learning the structure and function of tissues. If one wants to maximize the benefits of microscopy, it is essential to understand its capabilities and constraints. Different staining methods, including more recently, immunohistochemistry, have notably helped the capacity to recognize structures and certain cell types. The advancements in electron and fluorescence microscopy have also significantly increased the study of cell structure. In a nutshell, a microscope is a great tool for visualizing really tiny objects like microorganisms. 2.8 QUESTIONS 1. List down any FIVE (5) types of microscopes. ❖ Fluorescence Microscope ❖ Electron microscope ❖ Phase Contrast Microscope ❖ Surgery Microscope ❖ Light Compound Microscope
2. List the components of light compound microscope and their function.
❖ Concave Mirror- Transmits or produces light to the optical system. ❖ Iris- Control amount of light that cross through the viewed object. ❖ Condenser/Diaphragm- Focus light onto the object. ❖ Stage- A space to place the object. ❖ Mechanical stage- To make the movement of the object easy. ❖ Magnification System- Joined in a tube which can be moved up or down by using the fine and coarse focus.
3. Why do we need to use immersion oil in order to observe (using 100x lens)? ❖ We need to use immersion oil in order to observe (using 100x lens) because the high magnification lens (100x) will cause light refractions.
4. Why the image observed is inverted?
❖ the image observed is inverted because the curvature of the first lens, which inverts the image.
5. What are the precautions of this experiment?
❖ The lens (100x) should be cleaned up right after use. ❖ Do not touch the glass part of the lenses with your fingers. Use only special lens paper to clean the lenses. ❖ Always carry a microscope with both hands. Grasp the arm with one hand and place the other hand under the base for support. 2.9 REFERENCES
1. Lecture Notes chapter 2: Microscopy
https://drive.google.com/file/d/1w0STCSdi5hTdcIwHBX12rCs- S76cOjGO/view?pli=1 2. Biology Practical 1 Manual https://drive.google.com/file/d/1Ku6CSjaBpHuVBczgQzJ2oATfspRALdc4/vie w 3. Microscopy experiment video reference by lecturer https://drive.google.com/file/d/1JFhsboPRw-Kfm8-juWLaxLoCq7FNXo_- /view 4. Microscopes and how to use a light microscope https://www.youtube.com/watch?v=tVcEEw6qbBQ
