Human Biology BIOL-104 Laboratory Manual 2024-2025 PDF

Human Biology BIOL-104 Laboratory Manual LABORATORY 1 1.1 LAB SAFETY AND EQUIPMENT All students must read and understand the information in this document with regard to laboratory safety and emergency procedures prior to the fir...

Human Biology BIOL-104 Laboratory Manual LABORATORY 1 1.1 LAB SAFETY AND EQUIPMENT All students must read and understand the information in this document with regard to laboratory safety and emergency procedures prior to the first laboratory session. Your personal laboratory safety depends mostly on YOU. Effort has been made to address situations that may pose a hazard in the lab, but the information and instructions provided cannot be considered all-inclusive. Students must adhere to written and verbal safety instructions throughout the academic term. Since additional instructions may be given at the beginning of laboratory sessions, it is important that all students arrive at each session on time. With good judgment, the chance of an accident in this course is very small. Nevertheless, research and teaching workplaces (labs, shops, etc.) are full of potential hazards that can cause serious injury and/or damage to the equipment. Working alone and unsupervised in laboratories is forbidden if you are working with hazardous substances or equipment. With prior approval, at least two people should be present so that one can shut down equipment and call for help in the event of an emergency. Biology Unit, Imam Abdulrahman Bin Faisal University 6 Human Biology BIOL-104 Laboratory Manual 1.1.1. EMERGENCY RESPONSE It is your responsibility to read safety and fire alarm posters and follow the instructions during an emergency Know the location of the fire extinguisher, eye wash, and safety shower in your lab and know how to use them. Notify your instructor immediately after any injury, fire or explosion, or spill. Know the building evacuation procedures Biology Unit, Imam Abdulrahman Bin Faisal University 7 Human Biology BIOL-104 Laboratory Manual 1.1.2. PERSONAL AND GENERAL LABORATORY SAFETY 1. Never eat, drink, or smoke in the laboratory. 2. Read labels carefully. 3. Only trained and approved persons can use laboratory equipment. 4. Clothing– Wear laboratory coats. When handling dangerous and toxic substances, wear gloves, and safety shield or glasses. Shorts and sandals should not be worn in the lab. Coats should be hung in the hall or placed in a locker. 5. Long hair and loose clothes must be tied back or confined. 6. Keep the work area clear of all materials except those needed for your work. Don’t block air flow from equipment to prevent overheating. 7. Disposal– Students are responsible for the proper disposal of used materials in the appropriate containers. 8. Equipment Failure– If a piece of equipment fails, report it immediately to your lab assistant or tutor. Never try to fix the problem yourself. 9. If leaving a lab unattended, turn off all ignition sources and lock the doors. Biology Unit, Imam Abdulrahman Bin Faisal University 8 Human Biology BIOL-104 Laboratory Manual 1.1.3. BIOLOGICAL HAZARDS Biological hazards are organic substances that pose a threat to the health of humans and other living organisms. They include pathogenic micro-organisms, viruses, toxins (from biological sources), spores, fungi, bio-active substances and biological vectors or transmitters of diseases. Workers in health care professions are exposed to biological hazards via contact with human bodily matter, such as blood, tissues, saliva, mucous, urine and feces, because these substances have a high risk of containing viral or bacterial diseases. Likewise, people who work with live animals or animal products (blood, tissue, milk, eggs) are exposed to animal diseases and infections, some of which (zoonoses) have the potential to infect humans (e.g. Q-fever, avian flu or Hendra virus) or cause serious allergy via sensitization. Exposure to biological hazards can also occur when people are in contact with laboratory cell cultures, soil, clay and plant materials, organic dusts, food, rubbish, wastewater and sewerage. Exposure to molds and yeasts is common in some industrial processes, in workplaces with air conditioning systems and high humidity, and in the construction industry. Exposure to biological hazards is therefore widespread and the risk of exposure is not always obvious. Biology Unit, Imam Abdulrahman Bin Faisal University 9 Human Biology BIOL-104 Laboratory Manual 1.1.4. MICROSCOPES AND ACCESSORIES Compound microscope - Dissecting microscope - Glass slide – Coverslip – Slide box – Bottle of immersion oil - Dropper and dropper bottles - Lens paper Glass slide mount specimens Compound microscope Dissecting microscope for examination under a magnify small specimens magnify small dissected microscope (whole mount, or stained specimens sections) Bottle of immersion oil Coverslip Slide box Store immersion oil cover the specimen on a store and needed for examination with power 100X glass slide organize slides Dropper and dropper bottle Lens paper transfer small quantities of liquid clean microscope lenses (few drops) Biology Unit, Imam Abdulrahman Bin Faisal University 10 Human Biology BIOL-104 Laboratory Manual 1.1.5. DISSECTING MATERIALS Gloves - Dissecting pan - Dissecting pin – Probe – Scissors - Scalpel - Bone cutter – Forceps - Microbiological loop - Petri dish Dissecting pin Gloves Dissecting pan fix the specimen in place containment of specimen inside the dissecting pan. protection during dissection. Probe Scissors Bone cutter hold back organs to view cut specimens. cut small bones part of the specimen.. Scalpel Forceps cut specimens. hold part of specimen 1.1.6. MICROBIAL AND CELL CULTURE TOOLS Microbiological loop, petri dish. Microbiological loop microbial culture Petri dish Cell culture Biology Unit, Imam Abdulrahman Bin Faisal University 11 Human Biology BIOL-104 Laboratory Manual 1.1.7. OTHER EQUIPMENT Pipette- Balance (scale) – Hot plate – Stopwatch – water bath – spectrophotometer – pH meter - Centrifuge Pipette Digital Balance (scale) Hot plate measure definite weighing heating volume of liquids Spectrophotometer measure concentration of solutes by measuring the amount of light absorbed by the solution Centrifuge pH meter separation of fluids, particles, based measuring the pH on density by spinning Biology Unit, Imam Abdulrahman Bin Faisal University 12 Human Biology BIOL-104 Laboratory Manual NAME: ………………………………. ID #: …………………………………. GROUP: ……………………………... ACTIVITY 1.1. LAB SAFETY AND EQUIPMENT 1- True or false: a. It is prohibited to pipette any liquids by mouth. b. You should wear a lab coat all the time when working in the lab. c. Experienced students can work in the laboratory without supervision. Biology Unit, Imam Abdulrahman Bin Faisal University 13 Human Biology BIOL-104 Laboratory Manual NAME: ………………………………. ID #: …………………………………. GROUP: ……………………………... 2- Identify/name the following: a………………….. b………………….. c…………………….. d………………….. e………………….. Biology Unit, Imam Abdulrahman Bin Faisal University 14 Human Biology BIOL-104 Laboratory Manual NAME: ………………………………. ID #: …………………………………. GROUP: ……………………………... 3- Write the function of each of the following: a………………….. b………………….. c………………….. d………………….. e………………….. Biology Unit, Imam Abdulrahman Bin Faisal University 15 Human Biology BIOL-104 Laboratory Manual 1.2 LIGHT MICROSCOPY A microscope is a scientific instrument that magnifies small things to be seen better and examined correctly. There are many different types of microscopes. The most common are compound light microscope and electron microscope. In a compound light microscope, the object is illuminated: light is thrown on it. A compound microscope uses multiple lenses to create an enlarged image that is easier for a human eye to see. Materials: The compound microscope (ZEISS, OLYMPUS,… ) and microscope accessories. Methods: Students should work individually using compound microscopes and figures 2.1, 2.2 & 2.3 to learn the parts of the microscope, the function of each part and how to use it. Biology Unit, Imam Abdulrahman Bin Faisal University 16 Human Biology BIOL-104 Laboratory Manual Exercise 1.2.: Identify parts of the compound microscope and mention the function of each part. 17. Eyepiece (Ocular lens) 16. Binocular tube 2. Arm 15. Revolving nosepiece 14. Objective lens 12. Coarse focus control 10. Specimen holder 13. Fine focus control 9. Stage 6. Condenser assembly 8. Iris diaphragm 11. Specimen holder control 3. Light source 4. On-Off Switch 1. Base 5. Light source control Figure 1.1. Parts of compound microscope Biology Unit, Imam Abdulrahman Bin Faisal University 17 Human Biology BIOL-104 Laboratory Manual 18. Pointer 14. Objective lenses Red: 4X Yellow: 10X Blue: 40X White: 100X 8. Iris diaphragm 7. Condenser adjustment control 6. Condenser assembly Figure 1.2. Parts of compound microscope: Objective lenses (14), Pointer (18), and Condenser assembly (6) Biology Unit, Imam Abdulrahman Bin Faisal University 18 Human Biology BIOL-104 Laboratory Manual 17. Eyepiece lens 15. Revolving nosepiece 10. specimen holder Using oil with objective lens 100X 14. Iris diaphragm 7. Condenser adjustment control 6. Condenser Figure 1.3. Use of different parts of compound microscope Biology Unit, Imam Abdulrahman Bin Faisal University 19 Human Biology BIOL-104 Laboratory Manual 1.2.1. Parts of microscope and their functions 1. Base:the bottom of the microscope that has the light source. 2. Arm: the part that is connected to the base and helps to carry the microscope easily. One can hold the arm with one hand and put the other hand under the base of the microscope. 3. Light source:a small bulb, at the base of the microscope. 4. On-Off Switch: is an electrical power switch at the right side of the microscope base. It turns the illuminator on or off. 5. Light source control: adjusting this control increases or decreases the amount of light traveling form the light source at base up to the stage. It has six settings, but it is usually set at 3 for use in our lab. 6. Condenser assembly: found under the stage and contains condenser lens that collects the light from the light source and focuses it on the specimen. 7. Condenser adjustment control: is a small knob on the left under the stage. The condenser can be raised or lowered by this knob to allow for optimum illumination of the specimen. 8. Iris diaphragm: part of the condenser assembly used to control the amount of light reaching the specimen. In a student scope it is opened and closed by a rotating disk situated around the condenser. Rotating the disk clockwise increases the amount of light that goes through the condenser lens and rotating it anticlockwise reduces the amount of light passing through it. Biology Unit, Imam Abdulrahman Bin Faisal University 20 Human Biology BIOL-104 Laboratory Manual 9. Stage: is a flat surface where the slide with the specimen is placed. It has a central hole. Slightest movement using focus control knobs can affect the results. 10. Specimen holder: it holds the slide in place at the top of the stage. 11. Specimen holder control: attached to the specimen holder are two knobs. On the OLYMPUS microscope, these knobs are on the right side of the stage. Turn each knob and see how the specimen holder moves. The upper large knob moves the specimen forward and backward while the lower small knob moves it from right to left or left to right. 12. Coarse focus controls: are pair of large knobs found on both sides of the microscope present on the lower region of the arm. The main function of this knob is to move the stage with the specimen up or down to adjust specimen slide in order to bring it to focus and show the best image possible. It should be only used with the scanning objective (4X). 13. Fine focus controls: a sub part of the coarse adjustment knob. It moves the stage slightly to bring the specimen into sharp focus. It can be used with all objectives. Biology Unit, Imam Abdulrahman Bin Faisal University 21 Human Biology BIOL-104 Laboratory Manual 14. Objective lenses: usually there are four objective lenses in a standard microscope. Scanning Objective (4X): this shortest objective is useful for getting an overview of the slide. It is always safe to use as it cannot be lowered to the point of contacting and possibly breaking a slide. Low Power Objective (10X): this next shortest objective is probably the most useful lens for viewing slides. Almost any feature you need to observe in this course can be located with this objective lens. High Power Objective (40x): sometimes called the "high-dry" objective. It is useful for observing fine details such as the striations in skeletal muscle, the arrangement of Haversian systems in compact bone, types of nerve cells in the retina, etc. Oil Immersion Objective (100X): this longest objective is used for observing the details of individual cells such as white blood cells, the cells involved in spermatogenesis, etc. It must be used with a specially formulated oil that creates a bridge between the tip of the objective and the cover slip. Since the refractive indices of air and this lens are different, the lens will not work without this special oil. **Coarse focus control knob is safe to be used with objective lens 4X while fine focus control is used with other objective lenses. Biology Unit, Imam Abdulrahman Bin Faisal University 22 Human Biology BIOL-104 Laboratory Manual 15. Revolving nosepiece: the part of the microscope which holds the objective lenses. By turning clockwise or anticlockwise, the nosepiece revolves to switch the objective lenses in order to view the same specimen in various dimensions. 16. Binocular tube: is placed at the base of ocular lenses. Light passing through the objective lens is divided into two beams here. One beam goes to each eyepiece lens. 17. Eyepiece or ocular lens: this is where you position your eyes to observe the magnified image of the sample. It has a magnification of 10X. 18. Pointer: is located inside the eyepiece. It looks like a black line. Look through one eyepiece at a time and find the pointer. Total magnification: is calculated by multiplying the magnification of ocular lens (eyepiece lens) by magnification of objective lens. Table: Total magnification: *Complete the table. Objective lens Ocular lens Objective lens Total magnification magnification magnification Scanning power 4X 10 X 40 X Low power.…. X …. X ….. X High power ….. X …. X ….. X Oil immersion …… X …. X ….. X Biology Unit, Imam Abdulrahman Bin Faisal University 23 Human Biology BIOL-104 Laboratory Manual 1.2.2. How to use the compound microscope? Before using, clean the lenses (eyepiece, objective and condenser lens) with the lens paper. Using objective lenses 4X, 10X and 40X. 1. Turn the revolving turret so that the lowest power objective lens (4x) is clicked into position over the hole in the stage. 2. Place the microscope slide on the stage and fasten it with the specimen holder. Be sure the coverslip is facing upwards. 3. Adjust the condenser and light source to medium intensity. 4. Look at the objective lens 4X and the stage from the side and turn the focus knob so the stage moves upward. Move it up as far as it will go. 5. Look through the eyepiece and move the microscope slide around using specimen holder control until the sample is in the centre of the field of view. 6. Move the coarse focus knob until the image comes into focus. 7. Use the fine focus knob to place the sample into focus and readjust the condenser and light intensity for the clearest image (with low power objectives you might need to reduce the light intensity or shut the condenser). 8. When you have a clear image of your sample with the lowest power objective, you can change to the next objective lenses 10X or 40 X. You might need to readjust the sample into focus by using fine focus control and/or readjust the condenser and light intensity. Do not let the objective lens touch the slide! Biology Unit, Imam Abdulrahman Bin Faisal University 24 Human Biology BIOL-104 Laboratory Manual Using objective lens 100X (oil immersion lens): 1. To use the objective lens 100X (oil immersion), first make sure the specimen is in focus under the high power (40X) objective. 2. Move the high power objective out of position, place a small drop of oil on top of the cover slip above the specimen to be viewed and move the oil immersion lens into place as in Figure 1.3. 3. Use the fine focus adjustment knob to bring the specimen into focus. 4. When you are finished, click the low power lens (4X) into position and remove the slide. 5. Make sure to clean the lens and slide with lens paper. **It is extremely important that the oil does not contact any of the other objectives. If this happens, clean the lenses immediately. 6. Switch off the light and cover the microscope. Biology Unit, Imam Abdulrahman Bin Faisal University 25 Human Biology BIOL-104 Laboratory Manual NAME: ………………………………. ID #: …………………………………. GROUP: ……………………………... ACTIVITY 1.2: COMPOUND MICROSCOPE 1. What adjustment focus control knob is used with 40X lens? 2. Which part(s) of microscope controls the amount of light? 3. What are ocular lenses? 4. Which objective you should be used to get 40X total magnification? 5. True or false: a. Immersion oil must be used with the 10X objective lens. b. The stage is where the slide with the specimen is placed. c. The ocular lens is where you position your eyes to observe the specimen. It has no effect on magnification. 6. Write the name of the labeled parts of this microscope: D A E B F C Biology Unit, Imam Abdulrahman Bin Faisal University 26 Human Biology BIOL-104 Laboratory Manual LABORATORY 2 CELL STRUCTURE The aim of this lab is to identify the components of prokaryotic and eukaryotic cells particularly eukaryotic animal cells. Materials: Cell models of the prokaryotic and eukaryotic Cells. Electron micrographs of the components of prokaryotic (bacterial) cell and eukaryotic (plant and animal) cells. Video presentation 2.1 of cell structure and function. Methods: 1. Observe the models and electron micrographs of the prokaryotic and eukaryotic cells and identify the different cell structures. 2. Watch the video presentation on the cell structure and function. 2.1. https://www.youtube.com/watch?v=URUJD5NEXC8 3. After your observations, complete activity 2.1. Biology Unit, Imam Abdulrahman Bin Faisal University 27 Human Biology BIOL-104 Laboratory Manual 2.1. Comparison between eukaryotes and prokaryotes Eukaryotic Cells Prokaryotic Cells Have true nucleus; DNA is surrounded Have DNA region (nucleoid) by nuclear envelope. without nuclear envelope. Have membrane bound organelles. No membrane bound organelles. Ex: Protists, Fungi, Plants and Ex: Bacteria (Eubacteria and Animals Archaea bacteria) Figure 2.1. Eukaryotic and prokaryotic cells Biology Unit, Imam Abdulrahman Bin Faisal University 28 Human Biology BIOL-104 Laboratory Manual Cell wall Capsule Ribosome Plasma membrane Flagellum Cytoplasm Nucleoid Pilus (DNA) Plasma membrane Cell wall Nucleoid (DNA region) Cytoplasm with ribosomes Figure 2.2. Prokaryotic (bacterial) cell Biology Unit, Imam Abdulrahman Bin Faisal University 29 Human Biology BIOL-104 Laboratory Manual 2.2. Examples of Eukaryotic Cells Animal cell Plant cell No cell wall Has cell wall No chloroplasts (plastids) Contains chloroplasts No central large vacuole Has central large vacuole(s) Centriole is present Centriole is absent Some organelles of eukaryotic cells Membrane bound Non-membrane organelles bound structures Nucleolus Have double Have single Ribosomes membranes membrane Centrioles Cytoskeleton (Microtubules & Nucleus Microfilaments). Mitochondria Chloroplasts Endoplasmic reticulum Golgi bodies Vesicles - Lysosomes - Peroxisomes Biology Unit, Imam Abdulrahman Bin Faisal University 30 Human Biology BIOL-104 Laboratory Manual Central vacuole Cell wall Chloroplasts nucleus Figure 2.3. Eukaryotic (plant) cell Biology Unit, Imam Abdulrahman Bin Faisal University 31 Human Biology BIOL-104 Laboratory Manual nucleus Figure 2.4. Eukaryotic (animal) cell Biology Unit, Imam Abdulrahman Bin Faisal University 32 Human Biology BIOL-104 Laboratory Manual Nuclear envelope Nuclear pores chromatin nucleolus rough ER Chromatin Nucleolus Ribosomes Smooth Endoplasmic Rough Endoplasmic Reticulum (SER) Reticulum (RER) Figure 2.5. Nucleus and endoplasmic reticulum (ER) Biology Unit, Imam Abdulrahman Bin Faisal University 33 Human Biology BIOL-104 Laboratory Manual Cisterna Vesicle Figure 2.6. Golgi apparatus Outer Matrix Crista inner membrane membrane Figure 2.7. Mitochondrion Biology Unit, Imam Abdulrahman Bin Faisal University 34 Human Biology BIOL-104 Laboratory Manual Components of the cytoskeleton of the cell (microtubules and microfilaments) Nuclei (blue) Actin microfilaments (red) Microtubules (green) Figure 2.8. Cytoskeleton of the cell Biology Unit, Imam Abdulrahman Bin Faisal University 35 Human Biology BIOL-104 Laboratory Manual Cilia of Paramecium Flagellum of sperm Cilia of the respiratory tract doublet “9+2” array Flagellum of cilia and flagella Sperm Cilia Figure 2.9. Cilia and flagella Biology Unit, Imam Abdulrahman Bin Faisal University 36 Human Biology BIOL-104 Laboratory Manual Flagellum Plasma membrane Basal body triplet “9+0” array of centriole and basal body Basal body anchors flagellum Figure 2.10. Centriole and basal body Biology Unit, Imam Abdulrahman Bin Faisal University 37 Human Biology BIOL-104 Laboratory Manual NAME: ………………………………. ID #: …………………………………. ACTIVITY 2.1 GROUP: ……………………………... CELL STRUCTURE Mention the names of these organelles/structures D A B C E F G H Biology Unit, Imam Abdulrahman Bin Faisal University 38 Human Biology BIOL-104 Laboratory Manual 2.3. TONICITY AND OSMOSIS The cell membrane selectively permits some materials to cross. Under normal conditions, water constantly passes in and out of this membrane. Diffusion of water through a selectively permeable membrane is called osmosis. The selectively permeable membrane does not allow salt ions to pass into the cell. Water diffuses from a region of higher water concentration to a region of lower water concentration by osmosis until the transfer reaches equilibrium. Tonicity: the relative solute concentrations of two solutions – Hypotonic solution: a solution with greater water molecules and less solute molecules. – Isotonic solution: solutions with the same concentrations of water and solute molecules. – Hypertonic solution: a solution with less water molecules and more solute molecules If the cell is placed in hypotonic solution, water will move into the cell, the cell size increases, and it may burst. If the cell is placed in an isotonic solution, water will move into and out of the cell at the same rate and no change in cell size occurs. If the cell is placed in hypertonic solution, water will move from inside the cell to outside the cell. The cell shrinks as the cell loses water. Biology Unit, Imam Abdulrahman Bin Faisal University 39 Human Biology BIOL-104 Laboratory Manual Osmotic burst of blood cells (video) https://www.youtube.com/watch?v=OYoaLzobQmk S wollen RBCs (Hypotonic solution) Figure 2.12. Red blood cells in different solutions Experiment 2.1: Osmotic changes in potatoes In this experiment you will test the effects of diffusion and osmosis in potato cells. Procedure: Cut out two potato pieces ( ~ 1.5 x 1.5 inch). Using a scalpel (or vegetable peeler), make a cavity in the middle by scooping the soft tissue. Blot the cavity with tissue paper. Fill the cavity of one piece with salt. Leave the other piece empty. Record your observation in ~ 15 min. Explain your observation. Biology Unit, Imam Abdulrahman Bin Faisal University 40 Human Biology BIOL-104 Laboratory Manual NAME: ………………………………. ACTIVITY 2.2 ID #: …………………………………. TONICITY AND OSMOSIS GROUP: ……………………………... Answer the following questions regarding the effect of osmosis on red blood cells: A B C 1- What is the type of solution the above cells are immersed in? Solution A is:…………….……, explain why? …………………………………………………………………………………….. …………………………………………………………………………………….. Solution B is:……………….…, explain why? …………………………………………………………………………………….. …………………………………………………………………………………….. Solution C is:…………………., explain why? …………………………………………………………………………………….. …………………………………………………………………………………….. Biology Unit, Imam Abdulrahman Bin Faisal University 41 Human Biology BIOL-104 Laboratory Manual NAME: ………………………………. ID #: …………………………………. GROUP: ……………………………... ACTIVITY 2.3 TONICITY AND OSMOSIS Answer the following questions: What will happen to a plant cell that has a 0.8% salt concentration when it is placed into a solution containing a 10% salt concentration? Choose the correct answer: A) Water will move out of the plant cell, causing it to shrivel. B) Water will move in the plant cell, causing it to shrivel. C) Water will move in the plant cell, causing it to swell and burst. D) Water will move out of the plant cell, causing it to swell and burst. 2. Sally wants to get rid of some unwanted weeds, she decided to pour salt water on the ground where the weeds grow? Will this help Sally in removing the weeds? Why? --------------------------------------------------------------------------------------------------- --------------------------------------------------------------------------------------------------- --------------------------------------------------------------------------------------------------- --------------------------------------------------------------------------------------------------- --------------------------------------------------------------------------------------------------- Biology Unit, Imam Abdulrahman Bin Faisal University 42