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Biology Laboratory Manual-tenth Edition
Imaging System The imaging system improves resolution and magnifies the image. It consists of the objective and ocular eyepiece lenses and a body tube.
The objectives are three or four lenses mounted on a revolving nosepiece. Each objective is a series of several lenses that magnify the image, improve resolution, and correct aberrations in the image.
The most common configuration for student microscopes includes four objectives: Using the oil immersion objective requires special instructions, as explained in Exercise 24 to study bacteria. To avoid damaging your microscope do not use the oil immersion objective during this exercise. The magnifying power of each objective is etched on the side of the lens e. The ocular is the lens that you look through. Microscopes with one ocular are mon-. Oculars usually magnify the image ten times.
The body tube is a metal casing through which light passes to the oculars. In microscopes with bent body tubes and inclined oculars, the body tube contains mirrors and a prism that redirects light to the oculars. The stage secures the glass slide on which the specimen is mounted.
The viewing and recording system usually consists of a camera or video screen. Most student microscopes do not have viewing and recording systems. Basic Skills of Light Microscopy.
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Look through the oculars while using the coarse adjustment to focus on the specimen. Center the area of the specimen that you want to examine. Slowly rotate the high-power objective into place. Look through the oculars while you use the fineadjustment to focus on the specimen. For example, built-in light sources have replaced adjustable mirrors in the illuminating system, and lenses are made of better glass than they were in the past.
Your lab instructor will review with you the parts of the microscopes and their functions you will use in the lab. Procedure 3. Remove the microscope from its cabinet and carry it upright with one hand grasping the arm and your other hand supporting the microscope below its base. Place your microscope on the table in front of you. Question 1 a. As you view the letter e, how is it oriented? Upside down or right side up? How does the image move when the slide is moved to the right or left?
Toward you or away from you? Do not use paper towels or Kimwipes to clean the lenses of your microscope; they can scratch the lenses. Clean the lenses only with lens paper. Plug in the microscope and turn on the light source.
Always begin examining slides with the low-power objective. Locate the coarse adjustment knob on the side of the microscope. Only a partial turn of the coarse adjustment knob moves the stage or nosepiece a relatively large distance.
If your microscope is binocular, adjust the distance between the oculars to match the distance between your pupils. If your microscope is monocular, keep both eyes open when using the microscope. After a little practice you will ignore the image received by the eye not looking through the ocular. Focus a specimen by using the following steps: Place a microscope slide of newsprint of the letter e on the horizontal stage so that the e is directly below the low-power objective lens and is right side up.
It should be centered over the hole in the stage. Look through the oculars with both eyes open. Rotate the coarse adjustment knob i. Be sure that the e is directly below the objective lens and that you can see a spot of light surrounding the e.
Focus up and down to achieve the crispest image. Adjust the condenser iris diaphragm so that the brightness of the transmitted light provides the best view. Do not use the oil immersion objective. To fine-focus the image. It was put into an ocular on your microscope so that the lines superimpose on the image focus an image on high power. Question 2 a. Ninth Edition 3. Be sure that the objective does not touch the slide!
If the objective does not rotate into place without touching the slide. Determine the Size of the Field of View Never use the coarse adjustment knob to fine- The field of view is the area that you can see through the ocular and objective fig. Turning this knob changes the specimen-to-objective distance slightly and therefore makes it easy to fine-focus the image. Ninth Edition Examine each objective and record the magnifications of the objectives and oculars of your microscope in table 3.
Knowing the size of the field of view is important because you can use it to determine the approximate size of an object you are examining. Slowly rotate the high-power i. Most light microscopes are parfocal. The circular. You may need to readjust the iris diaphragm because the high-magnification objective allows less light to pass through to the ocular..
Shown here is the letter e from newsprint that is magnified 40 times. Compare the size of the image under high magnification with the image under low magnification. Calculate and record in table 3. Most light microscopes are also parcentered. An ocular micrometer is a small glass disk with thin lines numbered and etched in a row. The field of view can be measured with ruled micrometers fig. How many times is the image of the e magnified when viewed through the high-power objective? Micrometers are used to calibrate microscopes and measure the size of specimens.
Before you can use the micrometer you must determine for each magnification the apparent distance between the lines on the ocular micrometer. Record the values here.
A stage micrometer is a glass slide having precisely spaced lines etched at known intervals. Align lines at the left edges 0 lines of the two micrometers by moving the stage micrometer fig. This means that you must calibrate the ocular micrometer by comparing its lines to those lines on a standard ruler called a stage micrometer. Count how many spaces on the stage micrometer fit precisely in a given number of spaces on the ocular micrometer. Calculate the distance in millimeters between lines of the ocular micrometer.
Calibrate the ocular micrometer for each objective on your microscope. The ruler cannot be used to measure the diameters of the field of view at medium and high magnifications because the markings are too far apart. Why is it more difficult to locate an object starting with the high-power objective than with the lowpower objective?
Using the low-power objective. The order of the threads will not be the same on all slides. Ninth Edition 5. Place the ruler on the stage and under the stage clips of your microscope. You can use this information in future labs as you measure the sizes of organisms and their parts. Use this information to determine the area of the circular field of view with the following formula: Which objective should you use to initially locate the specimen?
Determine the Depth of Field Depth of field is the thickness of the object in sharp focus fig. Depth of field varies with different objectives and magnifications.
Re-examine the threads using the high-power objective lens. The space between each line on the ruler should represent a 1-mm interval. If your microscope has a mechanical stage. Which provides the largest field of view. Record in table 3. Obtain a clear plastic ruler with a metric scale. Align the ruler to measure the diameter of the circular field of view.
Record your calculated FOV areas in table 3. Slowly focus with the coarse adjustment and then the fine adjustment until the metric markings on the ruler are clear. Also record for each objective lens in table 3. The Microscope Focus up and down and try to determine the order of the threads from top to bottom. Alternate Procedure 3. Carefully rotate the nosepiece to the objective of lowest magnification. Also calculate the radius.
Basic Skills of Light Microscopy Therefore. Calculate the radius. Ninth Edition For example. Is the same level of illumination best for all magnifications? Is the image always best with highest illumination? Your instructor will demonstrate this technique. Which objective. Air bubbles appear as uniformly round structures with dark. Experiment with various intensities of illumination. Sketch in the following field of view what you see. Examine your preparation of algae. The upper and lower layers of cells are out of focus.
Which magnifications require the most illumination for best clarity and contrast? Are all three colored threads in focus at low power? Use your calculations for the diameter of the field of view to estimate the length of the shrimp. Prepare a wet mount of some newly hatched brine shrimp Artemia.
Question 4 a. Can all three threads be in focus at the same time using the high-power objective? This fresh preparation is called a wet mount and can be viewed with your microscope. The fourth level should have the diaphragm completely open. Observe the image. Gently lower the coverslip into place with a dissecting needle. Ninth Edition Add a drop of algal culture to a clean microscope slide. If the protozoa are moving too fast for you to examine carefully. Approximately how long and wide is a brine shrimp?
Practice For practice using your microscope. Also examine the prepared slides available in the lab. Also prepare wet mounts of the cultures available in the lab and sketch the organisms that you see. Add a clean coverslip. The methylcellulose will slow the movement of the protozoa.
Why is it important to put a coverslip over the drop of water when you prepare a wet mount? See also figures 3. A dissecting microscope provides a much larger working distance than does a compound microscope. Dissecting microscopes are always binocular fig. What is the area of the field of view when you use the lowest magnification of your dissecting microscope? Use a ruler to measure the diameter of the field of view with your dissecting microscope at several levels of magnification.
This distance is usually several centimeters compared to a centimeter or less for a compound microscope. How does the image through a dissecting microscope move when the specimen is moved to the right or left? Use your dissecting microscope to examine the organisms available in the lab. This arrangement provides a three-dimensional image with a large depth of field.
This is in contrast to the image in a compound microscope. Question 8 What other differences are there between compound and dissecting microscopes? Each ocular views the specimen at different angles through one or more objective lenses. Use figure 3.
What is the area when you use the highest magnification? Sketch some of these organisms. Although a compound microscope can produce high magnifications and excellent resolution. Use the terms high. Place a microscope slide of the letter e on the stage. Specimens that can be observed with a compound microscope are limited to those thin enough for light to pass through them.
How does the direction of illumination differ in dissecting as opposed to compound microscopes? In contrast. As you view the letter e how is it oriented? Surface Areas. What are the shapes. Ninth Edition Figure 3. Establish a working lab group and obtain Investigation Worksheet 3 from your instructor.
Red blood cells. Red blood cells are filled with hemoglobin. Outline on Worksheet 3 your experimental design and supplies needed to test your hypothesis. Refer to your textbook or other book and describe how an electron microscope can resolve such small structures. Write a short essay about the advantages and limitations of a transmission electron microscope. Examine the micrograph of the letter e shown in figure 3.
What are the advantages of knowing the diameter of the field of view at a given magnification? Why must specimens viewed with a compound microscope be thin?
Why are they sometimes stained with dyes? What is the actual height of the letter? What is the function of each of the following parts of a compound and dissecting microscope? What is the importance of adjusting the light intensity when viewing specimens with a compound microscope?
Why is depth of field important in studying biological structures? How can it affect your ability to find and examine a specimen? The gametes are sperm in male animals and eggs in female animals.
In multicellular organisms. During nuclear division. Laboratory Manual to accompany Biology. In eukaryotic. These chromatids are called sister chromatids.
As we shall see. During interphase. Explain how the chromosome number is reduced. Explain why meiosis I is emphasized and not meiosis II. Explain how the chromosome number stays constant.
As a result of meiosis. In animals. Tenth Edition 8. Sexually reproducing organisms utilize another form of nuclear division. This laboratory examines both mitotic and meiotic cell division to show their similarities and differences and it also discusses chromosome abnormalities that can occur during mitosis and meiosis.
When the cytoplasm divides. State the event of each stage on the line provided: Figure 8. Table 8.
Investigators have also discovered that cytoplasmic organelle duplication occurs during interphase. Mitosis including cytokinesis. Because early investigators noted little visible activity between cell divisions.
Mitosis and Meiosis ph Mader: Immature cells go through a cycle that consists of four stages: But when they discovered that DNA replication and chromosome duplication occur during interphase. Tenth Edition Pr o 34 The cell cycle. S for synthesis. Using Figure 8. This illustration represents a chromosome as it would appear just before nuclear division occurs.
The spindle is a structure that appears and brings about an orderly distribution of chromosomes to the daughter cell nuclei. Counting the number of centromeres tells you the number of chromosomes in the models. When cell division is about to begin. Scanning electron micrograph of a duplicated chromosome. Label the sister chromatids. Drawing of a duplicated chromosome. The fact that plant cells lack centrioles suggests that centrioles are not required for spindle formation.
Animal Mitosis Animal Mitosis Models 1. Spindle Table 8. The parent cell is the cell that divides. Each species has its own chromosome number. The centrosome. Animal cells contain two barrel-shaped organelles called centrioles in each centrosome and asters.
DNA replication results in a duplicated chromosome that consists of two sister chromatids held together at a centromere. The blastomeres blastula cells shown are in different phases of mitosis see Fig. What is the number of chromosomes in each of the cells in this model series? Tenth Edition Mitosis Mitosis is nuclear division that results in two new nuclei.
Chromatin is condensing into chromosomes. The colors signify that the chromosomes were inherited from different parents. Centrosomes begin moving apart. Polar spindle fibers stretch from each and spindle is in process of forming. In this way. Chromosomes will become indistinct chromatin. Kinetochore spindle fibers attached to the sister chromatids come from opposite spindle poles. Telophase Daughter cells are forming as nuclear envelopes and nucleoli reappear.
Tenth Edition The phases of mitosis are shown in Figure 8. Mitosis is the type of nuclear division that 1 occurs in the body somatic cells. Examine a prepared slide of onion root tip cells undergoing mitotic cell division. Using high power. Plant cells do have centrosomes and this accounts for the formation of a spindle.
Identify the phases of plant cell mitosis using models of plant cell mitosis and Figure 8. In plants. Notice that plant cells do not have centrioles and asters. Plant Mitosis Plant Mitosis Models 1. You may be able to just make out the cell plate. In the boxes provided. Onion Root Tip Slide 1.
A contractile ring composed of actin filaments gradually gets smaller. Cytokinesis begins in anaphase. Scanning Electron Microscopy in Biology: How do you know? Tenth Edition Cytokinesis Cytokinesis.
A single cell becomes two cells by a furrowing process. During cytokinesis. Copyright by R. Kessel and C. Mitosis is cell division in which the chromosome number 3. Vesicles containing cell wall components fuse to form cell plate. If a parent cell has 16 chromosomes.
During cytokinesis in a plant cell. Answer this question. Were any of the cells of the onion root tip slide undergoing cytokinesis? In plant cells. The nuclei in the daughter cells have the cell had. The nucleus of the parent cell has the diploid 2n number of chromosomes—that is. Build another homologous pair of duplicated chromosomes using Figure Obtain the following materials: Experimental Procedure: Meiosis In this exercise.
One member of the pair will be red. Each chromatid will have eight beads. Before meiosis begins. Be sure to bring the centromeres of two units of the same color together so that they attract and link to form one duplicated chromosome.
Tenth Edition 41 Meiosis Meiosis is a form of nuclear division in which the chromosome number is reduced by half. Building Chromosomes to Simulate Meiosis 1. Be sure to bring the centromeres of two units of the same color together so that they attract.
Note that your chromosomes are the same as those in Figure The chromosomes of these homologous pairs are duplicated. Each chromatid will have 16 beads. The red chromosomes were inherited from one parent. Build a homologous pair of duplicated chromosomes using Figure 8. A diploid nucleus contains homologues. Homologues look alike and carry the genes for the same traits. After meiosis is complete. In sexually reproducing species.
Does DNA replication occur during interkinesis? Simulate crossing-over by exchanging the exact segments of two nonsister chromatids of a single homologous pair.
In some species. Why use nonsister chromatids and not sister chromatids? Metaphase I Position the homologues at the metaphase plate in such a way that the homologues are prepared to move apart toward the centrioles. At metaphase I. Separate the pairs of centrioles. Each new nucleus contains one from each pair of chromosomes. Synapsis is the pairing of homologues during prophase I.
During telophase I. What other combinations would have been possible? Alternate the colors at metaphase I. Simulate synapsis by bringing the homologues together. Prophase I 5. What combinations of chromosomes are at the poles?
Fill in the following blanks with the words red-long. Interkinesis Interkinesis is the period between meiosis I and meiosis II.
Homologues line up next to one another during a process called synapsis. In telophase I. Pole A: Crossing-over is an exchange of genetic material between two homologues. During prophase of meiosis I. It is a way to achieve genetic recombination during meiosis. During crossingover. During anaphase I. Telophase I 9. Anaphase I Separate the homologues. Place two pairs of centrioles outside the nucleus. Metaphase II Move the duplicated chromosomes to the metaphase plate.
What is another way that sexual reproduction results in genetic variation? Notice that meiosis II is exactly like mitosis except that the nucleus of the parent cell and the daughter nuclei are haploid.
What does this action represent? Telophase II Put the chromosomes—each having one chromatid—at the poles near the centrioles. A zygote contains the same number of chromosomes as the parent. Summary of Meiotic Cell Division 1. During metaphase II. Place two pairs of centrioles at opposite sides of these chromosomes to form the new spindle. Meiosis is cell division in which the chromosome number 3. During prophase of meiosis II.
Each chromosome attaches to the spindle individually. In reality both daughter nuclei go on to divide again. Whereas meiosis reduces the chromosome number. The parent cell has the diploid 2n number of chromosomes. How many chromosomes are at the metaphase plate? Anaphase II Pull the two magnets of each duplicated chromosome apart. In telophase II. Then completely fill the remaining volume in each tube with the yeast To test our hypothesis about yeast growth, we must design a suspension that is provided.
The experiment sug- gested by our specific question and hypothesis involves of- 6. For each tube, slide an inverted, flat-bottomed test tube fering sugar such as glucose to one population of yeast, down over the yeast-filled tube. Hold the yeast-filled offering protein to another population of yeast, and then tube firmly against the inside bottom of the cover tube measuring their respective growth rates.
Fortunately, yeast and invert the assembly. Your instructor will grows readily in test tubes. As yeast grows in a closed, anaer- demonstrate how to slip this slightly larger empty tube obic container it produces CO2 in proportion to how readily over the top of each yeast tube and invert the assembly.
CO2 production is easily measured If done properly, no bubble of air will be trapped at the by determining the volume of CO2 that accumulates at the top of the tube of yeast after inversion. A well-designed experiment 8. After 40 min measure the height mm of the bubble links a biological response to different levels of the variable be- of accumulated CO2.
Record your results in ing investigated. In this case, the biological response is CO2 Worksheet 1. The levels of the variable are 9. When you are finished, clean your work area and sugar and protein.
These levels are called treatments, and in dispose of the contents of each tube as instructed by our experiment they include glucose, protein, and a control. Not for distribution. The first calculation is the 1. Examine your raw data in Worksheet 1.
Calculate the mean of the response variable CO2 treatment, and for the control replicates.The sci center the national science ecology and can definitely see why! They have moved to the Medical Collection. Test your hypothesis. For the sake of ensuring complete and consistent communication to students, the college has identified a set of items that all course syllabi are expected to contain.
Natural variation occurs in all processes of biology.