MT-MLS105 Human Anatomy and Physiology with Pathophysiology Lecture 2: The Microscope PDF

HUMAN ANATOMY AND PHYSIOLOGY WITH PATHOPHYSIOLOGY Laboratory Joey B. Asuncion Katterina Kleine Macaraig Vanessa Mae B. Moscoso BSMLS 1-2...

HUMAN ANATOMY AND PHYSIOLOGY WITH PATHOPHYSIOLOGY Laboratory Joey B. Asuncion Katterina Kleine Macaraig Vanessa Mae B. Moscoso BSMLS 1-2 Lecturer LECTURE 2: THE MICROSCOPE 16th Century - Zacharias Janssen An eyeglass maker experimented with OUTLINE ! multiple lenses which were placed in a History tube. Types of Microscope Important Parts of a Compound In spite of being the first one who used Microscope the microscope, he is not the father of Types of Brightfield Illumination microscopy. Total Magnification Proper Use of the Microscope Things to Remember a. Difficulty in focusing the specimen? b. The specimen became blurry when moved to a higher PO. c. The following checklist can aid in identifying the problem. MICROSCOPE - is an optical instrument designed to produce magnified visual or 17th Century - Anton Van photographic images of small images. Leeuwenhoek (ex: cells) Is a Dutch scientist and is the Father of Microscopy. He used the Simple With the invention of the microscope, Microscope (single lens). scientist or the biologist, we’re able to use this tool to observe and study the He is the first person to see and structure of small objects or describe bacteria, protozoa, yeast, and microorganisms. sperm. HISTORY ! 1000 AD - Reading Stone Or can be known as rock crystal or glass sphere. This is the first vision instrument—This is placed on top of the - Robert Hooke text to magnify the letters. The first to use the basic microscope with 2 converging lens systems or also known as the Compound Microscope. He wrote and illustrated a book called the “Micrographia”. He was also the first man to discover the cellular structure of cork. ck, dlsmhsi bsmls ‘28  HUMAN ANATOMY AND PHYSIOLOGY WITH PATHOPHYSIOLOGY Laboratory Joey B. Asuncion Katterina Kleine Macaraig Vanessa Mae B. Moscoso BSMLS 1-2 Lecturer LECTURE 2: THE MICROSCOPE Uses two lens systems to magnify the image. (Has an eyepiece and objective lens) Light illuminated. Condenser lens is present. Can have up to 4 objective lenses of different magnifications. 1930s - Frits Zernike Invented the Phase-Contrast Microscope that is used for the study of colorless and transparent biological materials. 1931 - Ernst Ruska and Max Knoll Invented and developed the Electron 3. Dissecting Microscope Microscope in Germany. Also known as “Stereoscopic Microscope or Stereoscope”. The image that appears is TYPES OF MICROSCOPE ! three-dimensional. Used to study the surfaces of the solid 1. Simple Microscope specimens. Uses a single lens for magnification. Natural light is the source to see the object. The condenser lens is not present. Non-adjusting magnification (only has one magnification). 4. Phase Contrast Microscope A contrast-enhancing optical technique. An instrument that permits the study of internal cell structure without the need to stain. Used to count platelets. 2. Compound Microscope Most commonly used. ck, dlsmhsi bsmls ‘28  HUMAN ANATOMY AND PHYSIOLOGY WITH PATHOPHYSIOLOGY Laboratory Joey B. Asuncion Katterina Kleine Macaraig Vanessa Mae B. Moscoso BSMLS 1-2 Lecturer LECTURE 2: THE MICROSCOPE Living specimens are destroyed by the high radiation of electron beams. 5. Polarized Light Microscope A contrast-enhancing technique used to a. Scanning Electron Microscope identify substances such as crystals in (SEM) - Produces a 3-dimensional urine and other bodily fluids. image of the specimen’s surface features. (Has a magnification of 20,000x) b. Transmission Electron Microscope (TEM) - Produces a 2-dimensional image of a specimen. This also provides detailed structure of the internal ultrastructure of the cells. (Has a magnification of 20,000,000x) 6. Dark Field Microscope Employs a special condenser. IMPORTANT PARTS OF A COMPOUND Used for the demonstration of very thin MICROSCOPE ! bacteria. Helpful in microbiology in the 1. Eyepiece - also known as ocular. It identification of spirochetes. magnifies the image by the factor The background will be seen as black indicated on the ocular’s barrel. (Has a instead of color. magnification of 10x) 2. Body tube - it holds the ocular in place. 3. Arm - the entire upper assembly of the microscope is held in an upright position by a bar called the arm. 4. Nosepiece - allows for easy rotation from one objective lens to another. 5. Objective lenses - for magnification. It has different classifications in four different colors: red, yellow, blue, and black. 6. Stage - supports the prepared 7. Electron Microscope microscope slide. Uses a beam of electrons as a source of 7. Stage clips - to hold the specimen in illumination. place. ck, dlsmhsi bsmls ‘28  HUMAN ANATOMY AND PHYSIOLOGY WITH PATHOPHYSIOLOGY Laboratory Joey B. Asuncion Katterina Kleine Macaraig Vanessa Mae B. Moscoso BSMLS 1-2 Lecturer LECTURE 2: THE MICROSCOPE 8. Stage adjustment knob - this is used to TOTAL MAGNIFICATION ! adjust the stage clip. 9. Condenser lens - focuses light onto the specimen. 10. Coarse Adjustment Knob - adjusts the distance between the stage. 11. Fine Adjustment Knob - adjusts the stage with a very little distance. 12. Base - it is the scope that holds or supports the whole microscope. 13. Lamp - this can be found below the stage. This is a high intensity lamp and the most common source or lamp source used in a microscope is tungsten 1. Scanning Objective - for initial location of halogen light bulbs. the specimen. 14. Diaphragm - this is used to change the 2. Low Power Objective - also used for initial intensity of light. location of the specimen and used for observing specimens that don’t need greater Two Types of Diaphragm: magnification. a. Disk Diaphragm - It has a rotating disc with different holes and diameters. This 3. High Power Objective - used for specimens also adjusts the light that passes that need greater magnification and also through the specimen. called “dry objective”. Dry Objective - does not require oil to b. Iris Diaphragm - It is made of visualize specimen overlapping silver and metal in a circular pattern resembling the iris of a human eye. 4. Oil Immersion Objective - used for magnification of extremely small specimens such as bacteria. Must be immersed in oil—Reason why it was called Oil Immersion TYPES OF BRIGHTFIELD ILLUMINATION ! Objective. Immersion oil is required to increase the refractive index when 100x of oil 1. Critical Illumination - The light source immersion objective lens is used. is focused on the specimen. The light that passes through the glass slide is uneven or not equal but it is focused on the specimen. PROPER USE OF THE MICROSCOPE ! 2. Koehler (Köhler) Illumination - The Place the microscope on a clean, flat light source is focused at the surface. condenser aperture diaphragm and Always carry the microscope with two the light is evenly distributed—Most hands: One under the base and the commonly used in a microscope. other grasping the arm. Unwind the cord from the back of the microscope and connect it to the power supply. ck, dlsmhsi bsmls ‘28  HUMAN ANATOMY AND PHYSIOLOGY WITH PATHOPHYSIOLOGY Laboratory Joey B. Asuncion Katterina Kleine Macaraig Vanessa Mae B. Moscoso BSMLS 1-2 Lecturer LECTURE 2: THE MICROSCOPE Make sure the stage and the objective Clean the objective lens and other are at the farthest distance and that the surfaces. lowest-power objective is in position. Turn on the microscope. Open the Diaphragm. THINGS TO REMEMBER ! Make sure that the light intensity dial is on the lowest setting. When not in use for an extended period Place the slide in place with the 2 stage of time, the microscope should be clips. covered or protected from dust. Use the stage adjustment knob to Before and after use, inspect the center the specimen over the condenser component parts. lens. Avoid placing fingers on the lens’ With a binocular eyepiece, turn the surface. eyepiece to find the correct space DO NOT use water to clean the lenses. between the eyes, or interpupillary distance. DIFFICULTY IN FOCUSING THE Start focusing on the lowest power SPECIMEN ? objective lens (4x). Use the coarse adjustment knob to Make sure the specimen is centered. move the objective as close as possible Start over at the lowest power and to the slide. move the stage back to its highest Look through the eyepiece and adjust position. the lamp controller or diaphragm for the greatest amount of light. THE SPECIMEN BECAME BLURRY WHEN Slowly turn the coarse adjustment so MOVED TO A HIGHER POWER LENS that the objective lens goes up. When the image becomes clear, switch Clean the lens! to the fine adjustment knob to make the image even sharper. THE FOLLOWING CHECKLIST CAN AID IN Magnify the image with a higher IDENTIFYING THE PROBLEM ! objective. Use the fine adjustment knob when working with the higher objectives. Oculars - Clean? Securely assembled? Magnify the image using the Oil Objective lens - Screwed in tightly? Dry Immersion Objective Lens. objective free oil? 1. Rotate the nosepiece halfway to Condenser - adjusted to proper height? the OIO lens. Free of oil? 2. Put a drop of immersion oil on Slide - correct side up? the slide, and then rotate the OIO Coverslip - Does it only have one lens into the oil drop. coverslip on slide? When finished, rotate the lowest-power Light source - Fingerprint on bulb? Bulb objective lens (4x) in viewing position in need of changing? Light source and make sure that the objective and aligned correctly? stage are as far apart as possible. Remove the slide. Turn off the light and power and wrap the cord. ck, dlsmhsi bsmls ‘28