Pasteur Experiment

Recreation of Pasteur’s Experiment

Introduction:

Today, we take many things in science for granted. Many experiments have been performed and much knowledge has been accumulated that people didn’t always know. For centuries, people based their beliefs on their interpretations of what they saw going on in the world around them without testing their ideas to determine the validity of these theories — in other words, they didn’t use the scientific method to arrive at answers to their questions. Rather, their conclusions were based on untested observations.

Among these ideas, for centuries, since at least the time of Aristotle (4th Century BC), people (including scientists) believed that simple living organisms could come into being by spontaneous generation. This was the idea that non-living objects can give rise to living organisms. It was common “knowledge” that simple organisms like worms, beetles, frogs, and salamanders could come from dust, mud, etc., and food left out, quickly “swarmed” with life. For example:

Observation: Every year in the spring, the Nile River flooded areas of Egypt along the river, leaving behind nutrient-rich mud that enabled the people to grow that year’s crop of food. However, along with the muddy soil, large numbers of frogs appeared that weren’t around in drier times. Conclusion: It was perfectly obvious to people back then that muddy soil gave rise to the frogs.

Objective:

In this experiment, you will conduct an experiment similar to the one done by Pasteur whenever he disproved spontaneous generation.

 

Materials Needed:Experiment Set-Up

  • Low-salt broth (chicken or beef, home-made or purchased)
  • 2  250-mL Erlenmeyer flasks
  • 2  1-hole rubber stoppers with bent glass tubing inserted (see diagram)
  • Glycerine
  • Hot plate & pot holders
  • 50-ml Graduated Cylinder
  • Marker

Procedure:

  1. Students should work in teams of 2 to 3 people. Each team should perform the following steps.
  2. Use glycerine and a twisting motion to insert glass tubing into the stoppers. be sure to rinse off excess glycerine with water.
  3. Mark Erlenmeyer flasks accordingly:
    1. Flask 1 with stopper and glass tube going straight up
    2. Flask 2 with stopper and glass tube bent in S-curve
  4. Using a graduated cylinder, place about 50-mL of broth in each Erlenmeyer flask.
  5. Place appropriate lids on flasks.
  6. Use a hot plate to boil broth in flasks with appropriate lids on them for 30 min., then let cool.
  7. For the next ten days, observe the flasks and record any changes in color, turbidity, smell, etc. (Be careful to NOT remove the stoppers from the flasks.)

Data:

Microbial Growth Record
Record the appearance of the flask contents.

Day Flask 1 with Straight Tubing Day Flask 2 with S-shaped Tubing
1 1
2 2
3 3
4 4
5 5
6 6
7 7
8 8
9 9
10 10

Conclusion:

  1. What was the appearance on the broth in each flask on Day 1?
  2. Was their an observed appearance change in flask 1 over the 10 days? Describe the change, if any.
  3. Was their an observed appearance change in flask 2 over the 10 days? Describe the change, if any.
  4. Explain why there was or was not a change in the appearance of the broth in each flask.
  5. Why do you think the idea of spontaneous generation was believed to be true for so long (1000+ years)?
  6. Did your experiment support spontaneous generation of organisms? Explain why or why not?

Osmosis & Diffusion in Egg Lab

 

Osmosis & Diffusion in an Egg

 

Objective:
In this investigation, you will use a fresh hen’s egg to determine what happens during osmosis & diffusion across membranes.

Materials: (per lab group)
1-2 fresh hen eggs in their shells, masking tape & marker, distilled water, clear sugar syrup (Karo, for example), vinegar, clear jar with lid, tongs, electronic balance, paper towels, paper, pencil

Procedure:

Day 1   

  1. Label the jar with your lab group & the word “vinegar”.
  2. Mass the egg with the electronic balance & record in the data table.
  3. Carefully place the raw egg into the jar & cover the egg with vinegar.
  4. Loosely re-cap the jar & allow the jar to sit for 24 to 48 hours until the outer calcium shell is removed.

Day 2   

  1. Open the jar & pour off the vinegar.
  2. Use tongs to carefully remove the egg to a paper towel & pat it dry.
  3. Record the size & appearance of your egg in your data table.
  4. Mass the egg on an electronic balance & record.
  5. Clean and re-label the jar with your lab group & the word “distilled water”.
  6. Carefully place the egg into the jar & cover the egg with distilled water.
  7. Loosely re-cap the jar & allow it to sit for 24 hours.

Day 3   

  1. Open the jar & discard the distilled water.
  2. Use tongs to carefully remove the egg to a paper towel & pat it dry.
  3. Record the size & appearance of your egg in your data table.
  4. Mass the egg on an electronic balance & record.
  5. Clean and re-label the jar with your lab group & the word “syrup”.
  6. Carefully place the egg into the jar & cover the egg with clear syrup.
  7. Loosely re-cap the jar & allow it to sit for 24 hours.

Day 4   

  1. Open the jar & pour off the syrup.
  2. Use tongs to very carefully remove the egg & rinse off the excess syrup under slow running water.
  3. Pat the egg dry on a paper towel.
  4. Record the size & appearance of your egg in your data table.
  5. Mass the egg on an electronic balance & record.
  6. Clean up your work area & put away all lab equipment.

Data:

 

RESULTS OF DIFFUSION

Original Mass Final Mass Appearance of Egg
VINEGAR
WATER
SYRUP

 

 

Questions & Conclusion:

1. Vinegar is made of acetic acid & water. Explain how it was able to remove the calcium shell.

 

2. (a) What happened to the size of the egg after remaining in vinegar?

(b) Was there more or less liquid left in the jar?

   (c) Did water move into or out of the egg? Why?

 

3. (a) What happened to the size of the egg after remaining in distilled water?

(b) Was there more or less liquid left in the jar?

   (c) Did water move into or out of the egg? Why?

 

4. (a) What happened to the size of the egg after remaining in syrup?

(b) Was there more or less liquid left in the jar?

   (c) Did water move into or out of the egg? Why?

 

5. Was the egg larger after remaining in water or vinegar? Why?

 

6. Why are fresh vegetables sprinkled with water at markets?

 

7. Roads are sometimes salted to melt ice. What does this salting do to the plants along roadsides & why?

 

 

 

 

Mitosis Activity

 

Stages of Mitosis

Introduction

Mitosis, also called karyokinesis, is division of the nucleus and its chromosomes.  It is followed by division of the cytoplasm known as cytokinesis.  Both mitosis and cytokinesis are parts of the life of a cell called the Cell Cycle.  Most of the life of a cell is spent in a non-dividing phase called Interphase.  Interphase includes G1 stage in which the newly divided cells grow in size, S stage in which the number of chromosomes is doubled and appear as chromatin, and G2 stage where the cell makes the enzymes & other cellular materials needed for mitosis.

Mitosis has 4 major stages — Prophase, Metaphase, Anaphase, and Telophase. When a living organism needs new cells to repair damage, grow, or just maintain its condition, cells undergo mitosis.

During Prophase, the DNA and proteins start to condense. The two centrioles move toward the opposite end of the cell in animals or microtubules are assembled in plants to form a spindle. The nuclear envelope and nucleolus also start to break up.


Prophase

During Metaphase, the spindle apparatus attaches to sister chromatids of each chromosome. All the chromosomes are line up at the equator of the spindle. They are now in their most tightly condensed form.


Metaphase

During Anaphase, the spindle fibers attached to the two sister chromatids of each chromosome contract and separate chromosomes which move to opposite poles of the cell.


Anaphase

In Telophase, as the 2 new cells pinch in half (animal cells) or a cell plate forms (plant cells), the chromosomes become less condensed again and reappear as chromatin. New membrane forms nuclear envelopes and the nucleolus is reformed.


Telophase

Objective: 

In this lab, you will determine the approximate time it takes for a cell to pass through each of the four stages of mitosis. You may use your textbook and class notes to help you identify the stages of mitosis as seen under the microscope. 

Materials:

Microscope, prepared slide onion root tip or whitefish blastula, textbook, lab worksheet, pencil

Procedure:

  1. Set up a compound light microscope and turn on the light.
  2. Place a slide containing a stained preparation of the Allium (onion root tip) or Whitefish blastula.
  3. Locate the meristematic or growth zone, which is just above the root cap at the very end of the tip or
  4. Focus in on low power, and then switch to medium or high power. Below find micrographs of the four stages of mitosis. Use them to help you identify the stages on the microscope slide.


Prophase (onion)

 


Metaphase (onion)

 


Anaphase (onion)

 


Telophase (whitefish)

 

  1. Now count the number of cells found in each stage of mitosis and place the data in the chart below.
  2. Determine the percentage of time each cell will spend in each stage of mitosis. Divide the number of each cell by the total number of cells and multiply by 100 to determine the percentage. Place these values in the chart below.

 

Stage of Mitosis Number of Cells Percent of time in each stage =

# of cells in stage     X  100%
Total # of Cell

Prophase %
Metaphase %
Anaphase %
Telophase %
Interphase
(Not a Mitotic Stage)
%
Total # cells 100%

 

  1. Line graph the data you have just collected.  Be sure to label the X and Y axis & include the units of measurement.

Title: __________________________________________________

Graph Legend:

 

Questions:

1. Of the four stages of mitosis, which one takes the most time to complete? 

 

2. Which is the shortest stage in duration?

 

3. What would happen if the process of mitosis skipped metaphase?  telophase?

Further Study:

Normal Cell Division may be observed in onion root tips. Many of the processes are similar to those in animal cells. However, in plant cells, the cell plate between daughter cells forms from the Golgi.

Find all of the stages of mitosis and  interphase in the above picture. Make a sketch of each stage and briefly describe what is occurring. Count and record the number of cells you see in each stage.

Projects
Notes

 

Metric System

Metric System (SI)
Scientist use a single, standard system of measurement.  The official name of the measurement system is SYSTEME INTERNATIONAL d’UNITES (International System of Measurements) or SI.

The metric system is based on the number 10.

Main Units of Measurement
Length Volume Mass
meter (m) liter (l) gram (g)

Using the above values (meter, liter, & gram) as the base, their value can be increased or decreased by moving the decimal point to the right (lowers the value) or left (raises the value).

Metric Conversion Table

Kilo-
(k)
Base Unit
(m, l, g)
Centi-
(c)
Milli-
(m)
Micro-
(µ)
nano-
(n)
x 1000 meter, gram, liter 100 1000 1000 1000
1000 1 .01 .001 .000001 .000000001

Convert the following values by moving the decimal point the correct number of spaces and in the right direction .

1. 69.8 meters (m) =  ________________ centimeters (cm)

2. 152.97 milliliters (ml)  =  ________________ liters (l)

3. 42.67 liters (l) = _____________ milliliters (ml)

4. 299.32 kilometers (km) = ____________ nanometers (nm)

5. 26 grams (g) = _____________ kilograms (kg)

6. 123.43 centigrams (cg) = ______________ grams (g)

7. 75.2 liters (l)  = __________________milliliters (ml)

8. 456.3 grams (g) = ________________ micrograms µg

9. 4507.22 kilometers (km) = _______________millimeters (mm)

10. 0.00297456 kilograms (kg) = ___________ nanograms (ng)

BACK

Mitosis and Meiosis

 

Mitosis and Meiosis

Introduction
All new cells come from previously existing cells. New cells are formed by the process of cell division which involves both replication of the cell’s nucleus (karyokinesis) and division of the cytoplasm( cytokinesis).

There are two types of nuclear division: mitosis and meiosis. Mitosis typically results in new somatic (body ) cells. Formation of an adult organism from a fertilized egg, asexual reproduction, regeneration, and maintenance or repair of body parts are accomplished through mitotic cell division. Meiosis results in the formation of either gametes (in animals) or spores ( in plants). These cells have half the chromosome number of the parent cell. You will study meiosis in Exercise 3B. Where does one find cells undergoing meiosis? Plants and animals differ in this respect. In higher plants the process of forming new cells is restricted to special growing regions called meristems. These regions usually occur at the tips of stems or roots. In animals, cell division occurs anywhere new cells are formed or as new cells replace old ones. However, some tissues in both plant and animals rarely divide once the organism is mature.

Exercise 3A.1: Observing Mitosis in Plant and Animal Cells Using Prepared Slides of the Onion Root Tip and Whitefish Blastula

Figure 3.1 Close up view of different stages of mitosis in an onion root tip:

 

 

Figure 3.2 Whitefish Blastula

 

Procedure:
Examine prepared slides of either onion root tips or whitefish blastula. Locate the meristematic region of the onion, or locate the blastula with 10X objective, and then use the 40X objective to study individual cells. Identify one cell which clearly represents each phase of mitosis. Sketch and label the cell in the box provided.

1. The non dividing cell is in a stage called interphase. The nucleus may have one or more dark-stained nucleoli and is filled with a fine network of threads, the chromatin. During interphase, DNA replication occurs.

Interphase

 

2. The first signs of cell division occurs in prophase. There is a thickening of the chromatin threads, which will continue until it is evident that the chromatin has condensed into chromosomes. With somewhat higher magnification you may be able to see the two chromatids held together by the centromere. As prophase continues , the chromatids continue to thicken and shorten. The nuclear envelope disappears and the beginnings of the spindle apparatus begin to appear.

 

Prophase

3. At metaphase, the chromosome pairs have moved to the center of the spindle. One particular part of each chromosome, the centromere, attaches to the spindle. The centromeres of all the chromosomes lie about the same level of the spindle called the metaphase plate.

 

r

Metaphase

4. At the beginning of anaphase, the centromere regions of each pair of chromatids separate and are moved by the spindle fibers toward opposite poles of the spindle, dragging the rest of the chromatid behind them. Once each chromatid is separate it is called a chromosome.

 

r

Anaphase

5. Telophase, the last stage of division, is marked by a pronounced condensation of the chromosomes, followed by the formation of a new nuclear envelope around each group of chromosomes. The chromosomes gradually uncoil into the fine threads of chromatin, and the nucleoli reappears. Cytokinesis may occur. This is the division of the cytoplasm into two new cells. In plants, a new cell wall is laid down between the daughter cells. In animal cells, the old cells will pinch off in the middle along a cleavage furrow to form two new daughter cells.

 

Telophase

 

Analysis Questions:
1. Why is it more accurate to call mitosis “nuclear replication” rather than “cellular division”?

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2. Explain why the whitefish blastula and onion root tip are selected for study of mitosis.

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Exercise 3A.2: Time for Cell Replication
Procedure:

It is hard to imagine that you can estimate how much time a cell spends in each phase of cell replication from a slide of dead cells. Yet this is precisely what you are going to do in this part of the lab. Since you are working with a prepared slide, you cannot get any information about how long it takes a cell to divide. What you can determine is how many cells are in each phase. From this, you can infer the percent of time each cell spends in each phase.

1. Observe every cell in one high power field of view and determine which phase of the cell cycle it is in. This is best done in pairs. The partner observing the slide calls out the phase of each cell while the other partner records. Then switch so the recorder becomes the observer and visa versa. Count at least two full fields of view. If you have not counted 200 cells, then count a third field of view.

2. Record your data in Table 3.1.

Table 3.1

Number of Cells

Percent of Total Cells Counted Time in Each Stage
Field 1 Field 2 Field 3 Total
Interphase
Prophase
Metaphase
Anaphase
Telophase
Total Cells Counted

3. Calculate the percentage of cells in each phase.

Consider it takes, on average, 24 hours (or 1,440 minutes) for onion root-tip cells to complete the cell cycle. You can calculate the amount of time spent in each phase of the cell cycle from the percent of cells in that stage.

Percent of cells in stage X 1,440 minutes = ___________ minutes of cell cycle spent in stage.

Questions:
1. If your observations had not been restricted to the area of the root tip that is actively dividing, how would your results have been different?

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2. Based on the data in Table 3.1, what can you infer about the relative length of time an onion root-tip cell spends in each stage of cell division?

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