DRAFT: This module has unpublished changes.

Abstract
The compound microscope requires skills necessary for understanding the makeup of organisms and how they react in certain environments. Once the observer fully understands how to work the microscope and can locate all of its parts with a knowledge of their functions, they will be able to know how to isolate different phases in an organism, as well as, viewing different specimens at different powers. Results may vary for each observer due to the differences in each slide produced or the microscope its self, but as for the onion root and the organisms' results they should be similar. Knowing how to use the compound microscope requires skill, which comes from practice. 


Introduction
This report presents an introduction to the compound microscope followed by a number of procedures. The purpose of a microscope is to see inside of organisms and cells- to see what is invisible to the naked eye [lab manual, 2014]. The microscope is marked by its accuracy and exactness.
The main objectives in this lab were the following: the microscope and all its functions had to be grasped, properly producing a wet mount slide to observe through the microscope, computing magnification, and knowing what to observe and describe. There were thirteen functions to be grasped, which included the arm, base, ocular lens, objective lens, nosepiece, body tube, stage, coarse adjustment knob, fine adjustment knob, light source, iris diaphragm, condenser, and observer. It was important to understand resolution and depth of field. Resolution is the shortest distance between two points that a user can still see as separate images under the microscope [eHow, 2009]. If the resolution is poor the image will appear blurry. Depth of field as it pertains to objects when viewed under a microscope is the vertical distance that remains in focus at one time [Lab Manual, 2014]. When preparing a wet mount it is important to have a clean microscope slide; a drop of water is added to the slide followed by a cover slip properly placed over the specimen. Another important component of this lab was computing magnification on a compound microscope. Magnification is represented with an "X" (times life size). For example, to find the total magnification of an object using the low power objective, which is 4X, multiply 10x4. 10 representing the eyepiece; 4 the low power objective. The total magnification would be 40X. The final area covered was locating an object and/or phase of an organism or specimen. In doing so it is important to understand what mitosis is in order to do the procedure. Mitosis is a type of cell division that results in two daughter cells each having the same number and kind of chromosomes as the parent nucleus, typical of ordinary tissue growth [Oxford Dictionary]. It is also important to know the distinct features of the following organisms: Amoeba, Euglena, and Paramecium, in order to properly view the live cultures. Amoeba do not like light, as well as, Paramecium, so the light on the microscope should be dimmed. Models and/or guided visuals can be a practical when trying to locate a certain object.

Materials and Methods
Procedure One: Using the Compound Microscope
In preparing the wet mount, a lower case letter e from the newspaper was carefully placed on a dry, clean slide. One drop of distilled water was added, followed by a clean cover slip placed on top. The slide was view on the microscope at a low power objective with adjustment from first the course adjustment then the fine for a clearer image. Effects of opening and closing the iris diaphragm were noted, as well as, appearance of specimen to the naked eye vs. through the microscope. The microscope was then returned to originally position and the slide was discarded in the glass container.
Procedure Two: The Importance of Light and Stain When Using a Bright Field Microscope
The following procedure consisted of two parts: an unstained specimen and a stained. A dry, clean slide was obtained and a drop of distilled water was added. Using a toothpick a sample of the lining of the cheek was placed on the distilled water and moved around to separate some of the cheek cells, followed by a cover slip on top. Once a single cheek cell was located, the high power objective was used to observe the cell. The second part of the procedure was a replication of the first, with the exception of a drop of methylene blue added before the cover slip was placed on top. The differences between the stained and unstained cheek cells were noted.
Procedure Three: Depth of Field
A prepared slide of three crossed colored threads were viewed with the low power objective in order to distinguish where the threads cross. While focusing slowly with the fine adjustment knob the effects observed were noted.
Procedure Four: Mitosis
A prepared slide of an onion root was observed using the high power objective once the meristematic region was located. The five stages of mitosis were located and recorded with visuals.
Procedure Five: Ameoba, Euglena, and Paramecia
In preparing the slide for Paramecia, a live Paramecia culture was transferred via pipette onto the slide, adding a small drop of Protoslo onto the culture. Observations made at low power to locate movement then high power if desired. Ameoba and Euglena were prepared the same way, without Protoslo added. Each organism was recorded with a visual and noted with a distinct feature.
Examples of each procedure can be found in the results section, each labeled according to the procedure. 


Results
Procedure One: Using the Compound Microscope
Question: Answer:
Is the ‘e’ that you are focusing on upside down or right side up as you look at it? Upside down
Now look through the microscope, is the ‘e’, when viewed through the microscope upside down or right side up? Right side up
Which way does the ‘e’ move when looking through the microscope while moving the top mechanical stage towards you? The ‘e’ moves to the right
Which way does the ‘e’ move when looking through the microscope while moving the bottom mechanical stage to your right? The ‘e’ moves to the left
Rotate the nosepiece to the high power objective and use the fine adjustment and iris diaphragm to focus clear on the ‘e’, does the ‘e’ appear as it did under the low power objective? No. The letter ‘e’ is larger and only one section of the ‘e’ can be seen

What is the total magnification of an object using the high power object?
40 times 10X = 400X

Procedure Two: The Importance of Light and Stain When Using a Bright Field Microscope
Question asked: Result:
Compare the ease of finding stain cheeks cell to that of finding unstained cheeks cells: The stained cheek cell was easier to view under the microscope light, because it made the specimen darker.

Procedure Three: Depth of Field
Three crossed colored threads: Results:
Top Thread: Red
Middle Thread: Blue
Bottom Thread: Yellow

Procedure Four: Mitosis
Interphase 40X Prophase 40X Metaphase 40X

Anaphase 40X Telophase 40X


Procedure Five: Ameoba, Euglena, and Paramecia
Ameoba 40X Euglena 40X Paramecium 40X 

Discussion
The purpose of this experiment was become familiar with using a compound microscope using five procedures. First, learning how to prepare a wet mount for the microscope was essential for all procedures. The first procedure consisted of preparing a wet mount with a lower case letter e from a newspaper to view with the microscope. The purpose of making this wet mount was to be able to view the effects the adjustment knobs, as well as, the iris diaphragm have on the specimen. When viewing the e on the stage it appeared to be upside down, but when looking through the microscope's eyepiece it appeared right side up, using low power moving the stage towards the observer the e appears to move away or when the stage is moved towards the right the e appears to move left when viewing through the eyepiece. The results concluded that the image through the microscopic view is opposite of the real image being emitted. In the second procedure, a sample of cheek cells was obtained by scrapping a toothpick along the cheek, placed on a wet mount, dislodged to separate the cells, and covered by a cover slip. The specimen was viewed on low power followed by high power focusing on a single cheek cell. The procedure for recreated for a second time, but methylene blue was placed on the wet mount before placing the cover slip on top. The results showed that using a stained cheek cell was easier to locate rather than the unstained. The third procedure consisted of viewing on low power a prepared slide of three crossed colored threads. The purpose of this experiment was to determine the order of the treads through a method called depth of field. The results showed that when the fine adjustment knob is moves up and down it causes one thread to be in focus; the others, blurry. The order of the threads: Top-red, Middle-Blue, and Bottom-yellow. Procedure four was viewing an onion root tip (meristematic region) to examine the phases of mitosis. The purpose of this experiment was to be able to correctly identify the five stages of mitosis: interphase, prophase, metaphase, anaphase, and telophase. The results for this experiment showed the characteristic of each phase. Interphase is the growth period, Prophase spindle fibers become visible and centrioles move to opposite sides of the cell, Metaphase chromosomes line up at metaphase plate and spindle fibers attach to centromeres of each chromatid pair, Anaphase chromatids separate and are pulled to opposite poles of the cell, and Telophase chromosomes are separated into masses. The last procedure consisted of preparing slides and viewing an Ameoba, Euglena, and Paramecia. The purpose of this procedure was to view the cellular features of the organisms. Protoslo was used on the Paramecia, but not on the Ameoba and Euglena. The results concluded that Euglena and Amoeba are slow moving, so protoslo is not necessary, whereas, Paramecia is fast moving. Amoeba does not like light; Paramecium does. Each organism has distinct features making them unique. These five procedures gave a greater understanding of how a compound microscope works, as well as, a visual of different organisms and their functions.  

 


Conclusion
The purpose of this lab was to learn about properly using a compound microscope. This was accomplished by viewing cellular reproduction mitosis and observing Ameoba, Euglena, and Paramecia. The five stages in mitosis were distinguished by being able to locate the meristematic region on the onion root where cells were undergoing mitosis when they were living. Observing the three organisms: Ameoba, Euglena, and Paramecia showed how every organism is unique and functions differently according to the environments they were created for. In conclusion, having an introduction to the compound microscope was essential in order to be able to view these specimens. It is the observer’s responsibility to use the microscope with care since it is a precision instrument and gives light on the objects we cannot see with the naked eye. 

Literature Cited
"Mitosis.”: Definition of in Oxford Dictionary (American English) (US). N.p., n.d. Web. 01 Dec. 2014.
Three Rivers Community College Staff. 2014. Human Anatomy & Physiology 1 Laboratory Exercises. Academx Publishing Services, Inc. pp.41-52
"What Is the Resolution of a Microscope?" EHow. Demand Media, 02 July 2009. Web. 17 Nov. 2014.

 

 

 

 

DRAFT: This module has unpublished changes.