Ford- Protein Crystallization

Protein Crystallization

with Standard Physical Science

and Honors Physical Science Students

Karen G. Ford, PhD

Ponte Vedra High School,

Ford- Protein Crystallization

Protein Crystallization with Standard Physical Science and Honors Physical Science Students

Karen G. Ford, PhD, Ponte Vedra High School

Abstract:

Entering high school freshmen in the St. Johns County school district who are not

recommended for honors science courses begin their science sequence with Physical Science.

Physical Science Honors students at Ponte Vedra High School are typically sophomores who

have completed Biology 1 Honors but were not recommended for Chemistry 1 Honors. The

course descriptions for Physical Science and Physical Science Honors are quite similar: they

include expectations that students evaluate the impact of biotechnology on individuals,

understand the concept of pH, and explore applications of the theory of electromagnetism. I

propose to evaluate and compare the understanding of protein crystallography and its

applications for biomedical research in physical science and physical science honors students.

Specifically I propose to use the Lysozyme Crystallization protocol developed by Robert

McKenna, PhD, and evaluate students using multiple choice and short answer questions. In

addition I will assess student attitude towards biotechnology after completion of the lesson.

Rational:

Because the Florida State Department of Education course descriptions for Physical Science

(20003310) and Physical Science Honors (20003320) are extremely similar, while the student

populations are distinct, and because I will be teaching both courses for the first time during the

2010-2011 school year, I am interested in comparing learning outcomes in these two

populations of students following a lecture and hands-on laboratory experience in protein

crystallography. I hypothesize that the Physical Science Honors students will perform better on

objective assessments of knowledge acquired during these activities because they have a

background in biology and are identified as “honors” students. However, these students were

not recommended to chemistry honors because they did not demonstrate above average

success in biology honors, so I wonder whether they have a bias against science that will hinder

their learning relative to standard physical science freshmen.

Dunham, et al. (2002) argued that biotechnology teaching should incorporate a blend of direct

instruction, cognitive problem solving, and constructivism. Consistent with their findings, I intend

to introduce the science of protein crystallography using a direct teaching approach. Next I will

instruct students on the laboratory procedures, and encourage them to use their cognitive

abilities to implement the protocol and modify it when appropriate. Finally, I will use video and

anecdotal clinical studies from Dr. Robert McKenna’s presentation to assist students as they

construct their own appreciation for this technology’s applications to medicine.

Dawson and Soames (2006) used a pre-test post-test scenario and observed that, after a 10-

week course in biotechnology, students in Australia demonstrated increased understanding of

the technology, but little change in their attitude towards applications of biotechnology, with the

exception of their increased acceptance of medical applications of biotechnology. The

researchers were not able to discern a trend in individual student’s responses to questions

Ford- Protein Crystallization

about the morality of biotechnology before and after completing the biotechnology curriculum. I

will also evaluate both understanding of and attitudes towards biotechnology in my research.

Dawson (2007) surveyed 12-17 year old students with regards to their understanding and

approval of various biotechnology applications, and determined that older students generally

responded more favorably towards biotechnology than younger students. This research

involved interviews and surveys. I am interested to determine whether there is a significant

attitude difference in my two student populations following their biotechnology lesson. I

understand that there are many variables between these populations and the results will need to

be interpreted with caution.

The purpose of this study is to compare the effects of a lecture and a hands-on laboratory

experience in protein crystallization on two groups of students’ understanding of this

biotechnology application. Student understanding will be measured objectively using multiple

choice and short answer questions. A Likert scale will be used to assess attitudes towards

medical applications of biotechnology.

Action research intervention:

Protein Crystallization Lesson Plan.

Lesson Title: Protein Gems

Grade Span: 9th and 10th grade high school students

Content Emphasis: Physical Science and Biotechnology

Author: Karen G. Ford, PhD

School: Ponte Vedra High School

District: St. Johns County

Email: fordk@stjohns.k12.fl.us

Phone Number: (904) 547-7350

Learning Goals: This lesson will teach students a variety of kinesthetic and cognitive skills.

They will learn to

accurately pipette small volumes of solution using micropipettes,

collect data in a student-generated data table,

graph data,

interpret phase transitions using kinetic molecular theory,

identify variables that influence the formation of protein crystals,

Ford- Protein Crystallization

form rational hypothesis about the effects of pH on the structure of and formation of

crystals from lysozyme,

form rational hypothesis about the effects of salt concentration on the structure of and

formation of crystals from lysozyme,

explore applications of the theory of electromagnetism to x-ray crystallography, and

describe how protein crystallization can be used in rational drug design.

Estimated time: This lesson will require approximately 7 days of class time (45 minutes/day).

Day 1 – introduction to protein crystallography

Day 2 – introduction to micropipetting

Day 3 – set up protein crystallography lab

Day 4 – applications of protein crystallography to rational drug design

Day 5 – data collection and graphing

Day 6 – data interpretation and conclusions

Day 7 – qualitative and quantitative assessment of student learning

Materials and Resources:

This activity will require

“Clickers” and appropriate pre-test quiz to diagnose misunderstandings of pH, phase

transitions, and kinetic molecular theory.

Dr. Robert McKenna’s PowerPoint presentation to CPET Bench-to-bedside teachers on

June 24, 2010

Copies of the “Crystallization Hand on, Lysozyme crystallization” lab instruction.

Micropipettes and tips (P-1000,P-200 and P-20)

64 well dishes

Two colors of dye for practicing micropipetting

24-well Limbro plate

forceps

Ford- Protein Crystallization

Plastic micro-bridges for use in Limbro plates – 12 per student group

22 mm Round cover slips for covering wells

Vacuum grease and tooth picks for sealing cover slips on trays

Lysozyme at 50 mg / ml – on ice

ddH2O

4M NaCl

0.5M Sodium Acetate, pH 4.2

1M Tris-HCl, pH 7.0

Dissecting microscopes

Objective quiz to measure student learning

Likert quiz to measure student attitude towards medical biotechnology

Teacher Prepartion:

In preparation for this lesson, teacher should

Reserve a date with Kim for delivery of micropipetters, sterile pipette tips, and 64 well

dishes

Review Dr. McKenna’s PowerPoint presentation

Make copies of Lysozyme crystallization lab instructions

Make copies of micropipette practice lab instructions

Develop and makes copies of quiz and Likert survey

Lesson Procedure and Evaluation

Introduction:

I will introduce this lesson by reviewing concepts of pH, kinetic molecular theory, and phase

transitions, and I will assess student understanding with eInstruction (“clicker quizzes”). I will

attempt to clarify any misconceptions.

Ford- Protein Crystallization

I will ask students to brainstorm in small groups and propose ways this physical science

information could be used to improve human lives, how they think medical research scientists

identify new drugs to treat disease, if they have heard of drugs that lose their effectiveness, and

what they think the term “rational drug design” means. I will tell them that they will now engage

in an activity used by research scientists who are rationally designing pharmaceutical drugs.

Exploration: After teaching students to micropipette accurately, I will allow them to set up a 24

well Limbro plate containing varying concentrations of NaCl at two different buffered pH levels

(4 and 7). Students will then add lysozyme protein to sitting and hanging drops. After 4-7 days,

students will evaluate the results of the experiment and determine which combination of NaCl

and pH allowed the formation of large protein crystals.

Application: Students will be able to apply their knowledge in data collection and graphing to a

wide variety of laboratory activities both in this course and in future science courses. They will

be able to apply the technique of micropipetting to future labs in this course on PCR of genomic

DNA and electrophoresis of PCR-generated DNA fragments. Students will be taught

applications of protein crystallization technology to rational drug design using Dr. Robert

McKenna’s PowerPoint lecture given to teachers at the 2010-2011 Bench to Bedside summer

workshop.

Assessment: Students will be assessed with objective multiple choice and short answer

questions on the following SSS benchmarks:

MA.912.S.1.2 (Determine appropriate and consistent standards of measurement for the data to

be collected in a survey or experiment.)

MA.912.S.3.2 (Collect, organize, and analyze data sets, determine the best format for the data

and present visual summaries from the following: bar graphs, line graphs, stem and leaf plots,

circle graphs, histograms, box and whisker plots, scatter plots, and cumulative frequency

graphs).

SC.912.L.16.10 (Evaluate the impact of biotechnology on the individual, society, and the

environment, including medical and ethical issues).

SC.912.N.1.1 (Define a problem based on a specific body of knowledge….. Basically use the

scientific method).

SC912.P.8.11 (Relate acidity and basicity to hydronium and hydroxyl ion concentration and pH).

SC,912.P.10.18 (Explore the theory of electromagnetism by comparing and contrasting the

different parts of the electromagnetic spectrum in terms of wavelength, frequency, and energy,

and relate them to phenomena and applications).

SC,912.P12.11 (Describe phase transitions in terms of kinetic molecular theory).

Ford- Protein Crystallization

Teacher Self-Reflection:

Connections to Bench to Bedside summer institute:

The protein crystallization technique used in this study will be nearly identical to that developed

by Dr. Robert McKenna for the Bench-to-Bedside summer institute for teachers through CPET

at University of Florida. We will utilize the technology made available by the CPET staff to

conduct the experiments and study the applications presented in Dr. McKenna’s PowerPoint

lecture on June 24, 2010.

Data Collection and Analysis:

Students will be divided into two groups: those enrolled in Physical Science (Group 1) and

those enrolled in Physical Science Honors (Group 2). Each group will be provided with the

same classroom experiences with regards to protein crystallization. Each group will be asked to

complete an object assessment of their knowledge after completion of the activities, and the

number of correct responses from each group will be compared. Data will be summarized by

mean, standard deviation, and analysis of variation statistics.

Literature cited:

Dawson, V. (2007) An Exploration of High School (12–17 Year Old) Students’ Understandings of, and AttitudesTowards Biotechnology Processes, Res Sci Educ (2007) 37:59–73. Dawson, V and C. Soames (2006) The effect of biotechnology education on Australian high school students' understandings and attitudes about biotechnology processes, Research in Science & Technological Education, 24: 2, 183-198. Dunham, T, Wells, J, and K. White (2002) Biotechnology Education: A Multiple Instructional Strategies Approach Journal of Technology Education Vol. 14 No. 1, p. 65-81. Budget and budget justification:

The activities proposed here require the use of the CPET micropipetters. The $200 equipment

will be applied towards the purchase of consumables including lysozyme, and 24-well Limbro

plates. Any remaining funds will be used to purchase student test tube racks and ice buckets.

Ford- Protein Crystallization

Assessment of student attitude towards biotechnology.

Please respond to the following items by drawing a circle around the response that most closely

reflects your opinion: strongly agree (SA), agree (A), undecided (U), disagree (D), or strongly

disagree (SD)

1. I feel good about my problem solving skills. SA A U D SD

2. My problem solving skills increased as a result of

this laboratory activity

SA A U D SD

3. My understanding of biotechnology has increased

as a result of this activity.

SA A U D SD

4. My interest in biotechnology has increased as a

result of this activity.

SA A U D SD

5. I enjoyed the experimental aspects of this activity. SA A U D SD

6. I want to learn more about jobs in the

biotechnology industry.

SA A U D SD

7. I am interested in pursuing a career in

biotechnology.

SA A U D SD

8. I understanding of the processes associated with

“rational drug design”.

SA A U D SD

9. I have an increased understanding of the ways that

mathematics can be used in biology.

SA A U D SD

10. Research on medical applications of biotechnology

should be funded by tax dollars.

SA A U D SD

Entering high school freshmen in the St. Johns County school district who are not recommended for honors science courses begin their science sequence with Physical Science.

Physical Science Honors students at Ponte Vedra High School are typically sophomores who have completed Biology 1 Honors but were not recommended for Chemistry 1 Honors.

The course descriptions for Physical Science and Physical Science Honors are quite similar

evaluate the impact of biotechnology on individuals,

understand the concept of pH, and

explore applications of the theory of electromagnetism

I propose to use the LysozymeCrystallization protocol developed by Robert McKenna, PhD, and evaluate students using multiple choice and short answer questions. In addition I will assess student attitude towards biotechnology after completion of the lesson.

The purpose of this study is to compare the effects of a lecture and a hands-on laboratory experience in protein crystallization on two groups of students’ understanding of this biotechnology application. Student understanding will be measured objectively using multiple choice and short answer questions. A Likert scale will be used to assess attitudes towards medical applications of biotechnology.

Estimated time: This lesson will require approximately 7 days of class time (45 minutes/day). Day 1 – introduction to protein crystallographyDay 2 – introduction to micropipettingDay 3 – set up protein crystallography labDay 4 – applications of protein crystallography to rational drug designDay 5 – data collection and graphingDay 6 – data interpretation and conclusionsDay 7 – qualitative and quantitative assessment of student learning

1. I feel good about my problem solving skills. SA A U D SD

1. My problem solving skills increased as a result of this

laboratory activity

SA A U D SD

1. My understanding of biotechnology has increased as a

result of this activity.

SA A U D SD

1. My interest in biotechnology has increased as a result of

this activity.

SA A U D SD

1. I enjoyed the experimental aspects of this activity. SA A U D SD

1. I want to learn more about jobs in the biotechnology

industry.

SA A U D SD

1. I am interested in pursuing a career in biotechnology. SA A U D SD

1. I understanding of the processes associated with

“rational drug design”.

SA A U D SD

1. I have an increased understanding of the ways that

mathematics can be used in biology.

SA A U D SD

1. Research on medical applications of biotechnology

should be funded by tax dollars.

SA A U D SD

 

 

 

 

A Study on the Use of Protein Crystallization  

with Standard Physical Science, Honors Physical Science, and Biology Honors Students  

 

 

 

 

Karen G. Ford, PhD 

Ponte Vedra High School,  

460 Davis Park Road,  

Ponte Vedra, Florida   32081 

      

 

 

Protein Crystallization with Standard Physical Science, Honors Physical Science, and Biology Honors Students  

Karen G. Ford, PhD, Ponte Vedra High School, 460 Davis Park Road, Ponte Vedra, Florida,  32081.       

Abstract:

Ponte Vedra High School freshmen enroll in one of three science courses depending on their math skills and teacher recommendations, with the highest performing students taking Biology Honors (BH), the middle group taking Physical Science Honors (PSH) and the lowest taking Physical Science (PS). Course descriptions for PS and PSH are quite similar, and include expectations that students evaluate the impact of biotechnology on individuals. Students from these three classes followed the Lysozyme Crystallization protocol developed by Robert McKenna, PhD. After completing the lesson, they were evaluated based on their success in growing crystals, their understanding of the technology associated with growing crystals as assessed by short responses to questions, and their attitude towards biotechnology. The PS students were mostly unsuccessful in growing crystals. PSH students were more successful, 61% grew crystals, and BH students were most successful (90%). Understanding and attitudes mirrored success for the three groups.

Rationale:

Because the Florida State Department of Education course descriptions for Physical Science (20003310) and Physical Science Honors (20003320) are extremely similar, while the student populations are distinct, and because I will be teaching both courses for the first time during the 2010-2011 school year, I was interested in comparing learning outcomes in these two populations of students following a lecture and hands-on laboratory experience in protein crystallography. I hypothesized that the Physical Science Honors students would perform better on objective assessments of knowledge acquired during these activities because they were identified as “honors” students by their middle school teachers. However, I predicted that the students taking the standard Physical Science course will have a more positive attitude toward biotechnology because they are typically not exposed to laboratory activities using innovative technology and the novelty may provide a more excitement.

My teaching assignment included one section of Biology Honors students that were co-enrolled in Biotechnology 1. Relative to the physical science students, these students had a stronger background in protein structure more more practice using micropipettes. I included these students in the study because of their pre-identified interest in biotechnology and because I expected they would follow the directions most accurately and thus be most likely to grow crystals even if the other groups failed.

Dunham, T., & White, K. (2002) argued that biotechnology teaching should incorporate a blend of direct instruction, cognitive problem solving, and constructivism. Consistent with their findings, I intend to introduce the science of protein crystallography using a direct teaching approach. Next I will instruct students on the laboratory procedures, and encourage them to use their cognitive abilities to follow the protocol and modify it when appropriate. Finally, I will use video and anecdotal clinical studies from Dr. Robert McKenna’s presentation to assist students as they construct their own appreciation for this technology’s applications to medicine.

Dawson, V., & Soames, C. (2006) used a pre-test post-test scenario and observed that, after a 10-week course in biotechnology, students in Australia demonstrated increased understanding of the technology, but little change in their attitude towards applications of biotechnology, with the exception of their increased acceptance of medical applications of biotechnology. The researchers were not able to discern a trend in individual student’s responses to questions about the morality of biotechnology before and after completing the biotechnology curriculum. I will also evaluate both understanding of and attitudes towards biotechnology in my research.

Dawson, V. (2007) surveyed 12-17 year old students with regards to their understanding and approval of various biotechnology applications, and determined that older students generally responded more favorably towards biotechnology than younger students. This research involved interviews and surveys. I am interested to determine whether there is a significant attitude difference in my two student populations following their biotechnology lesson. I

understand that there are many variables between these populations and the results will need to be interpreted with caution.

The purpose of this study is was to compare the effects of a lecture and a hands-on laboratory experience in protein crystallization on three groups of students’ understanding of this biotechnology application. Student understanding was measured objectively using short answer questions. A Likert scale was also used to assess attitudes towards medical applications of biotechnology.

Action research intervention:

Twenty- four ninth grade high school Physical Science students, eighty-one ninth grade high school Physical Science Honors students, and twenty-one ninth grade high school Biology Honors students attempted to follow a detailed protocol that resulted in formation of protein crystals. The Physical Science students had previously learned about phases of matter and were concurrently learning about chemical bonding. The Physical Science Honors students had previously learned about phases of matter, chemical bonding, and mixtures, and were concurrently learning about molecular interactions in solutions, and the Biology Honors students had learned about biochemistry, cells, genetics, and evolution, and were concurrently learning about ecology.

Physical Science, Physical Science Honors, and Biology Honors students were given a lecture on the techniques used for and applications of protein crystallization for 40 minutes on day 1. The lecture was accompanied by PowerPoint slides provided by Dr. Robert McKenna through the 2010 CPET Bench-to-Bedside summer course at the University of Florida. It included a description of protein structure and function, techniques used for crystallizing protein, a brief history of x-ray crystallography, and descriptions of two specific attempts at “rational drug design”.

Physical Science and Physical Science Honors students were then instructed in the proper use of micropipettes using a micropipetting lab protocol that was shared during the 2010 CPET Bench-to-Bedside summer course at the University of Florida. The lab allowed students to use P-200 and P-1000 micropipetters to accurately pipet designated volumes of two colored dyes into 96 well plates and form recognizable shapes and letters. Biology Honors students had experience using micropipetters in their biotechnology course, so they did not complete this activity.

The third day of the intervention required that follow a detailed protein crystallization lab protocol shared during the 2010 CPET Bench-to-Bedside summer course at the University of Florida. All of the students were expected to accurately pipette designated volumes of salt, buffers, and water into limbro plates, prepare drops of half-strength salt solution containing protein on coverslips or in microbridges, then seal the dishes with coverslips affixed by thin rings of immersion oil. Biology Honors students successfully set up 24 wells on their Limbro plates during the 45 minute class period. Many of the first section of Physical Science Honors students were not able to complete the set up of 24 wells during the class period. Midway through the class they were instructed to complete setting up as many wells as possible, but to be sure to add protein to drops for at least half of the plates. Subsequent sections of Physical Science Honors and Physical Science were instructed to only set up 12 wells.

After storing the plates for 5 days at room temperature, students evaluated their drops for protein crystal formation using stereomicroscopes with 20 and 40 X magnification. After first observing crystals, students were instructed to provide a short description of their results as follows:

1. Accurately sketch two protein crystals. 2. Describe how the lab encouraged proteins to form crystals in some of the drops. 3. Discuss why proteins might have crystallized in some of the salt concentrations but not in others. 4. Explain how protein crystals can be used for “rational drug design”.

Students completed a Likert-type survey that provided some insight into their attitude towards the laboratory experience and their general interest in biotechnology. Student responses to the objective questions and the survey were averaged and responses in the three groups were compared.

Time required. This lesson required 4 days of class time (39-45 minutes/day).

• Day 1 – introduction to protein crystallography and applications of protein crystallography to rational drug design

• Day 2 – introduction to micropipetting

• Day 3 – set up protein crystallography lab

• Day 4 – data collection and qualitative and quantitative assessments.

Materials and Resources:

• Dr. Robert McKenna’s PowerPoint presentation during 2010 CPET Bench-to-Bedside summer course at the University of Florida.

• Copies of the “Introduction to Micropipetting Lab” designed by CPET Bench-to-Bedside instructors and shared during 2010 CPET Bench-to-Bedside summer course at the University of Florida.

• “Crystallization Hand on, Lysozyme crystallization” lab instructions from CPET Bench-to-Bedside instructors and shared during 2010 CPET Bench-to-Bedside summer course at the University of Florida.

• Micropipettes and tips (P-1000,P-200 and P-20)

• 96 well dishes

• 6 colors of dye for practicing micropipetting

• 24-well Limbro plates generously provided by CPET at the University of Florida.

• forceps

• Plastic micro-bridges for use in Limbro plates generously provided by CPET at the University of Florida

• 22 mm -round cover slips for covering wells generously provided by CPET at the University of Florida

• Immersion oil in dropper bottles generously provided by CPET at the University of Florida

• Lysozyme at 50 mg / ml – on ice, generously provided by CPET at the University of Florida

• ddH2O

• 4M NaCl

• 0.5M Sodium Acetate, pH 4.2

• 1M Tris-HCl, pH 7.0

• Dissecting microscopes

• Short answer questions to assess student understanding

• Likert survey assessing attitudes towards medical biotechnology

Data Collection and Analysis.

Student data was initially divided into three groups: those enrolled in Physical Science, those enrolled in Physical Science Honors, and t hose enrolled in Biology Honors. However, after the micropipetting lab was marginally performed by the Physical Science students, then only one out of 10 groups in this cohort successfully grew protein crystals, their data was not further collected or analyzed.

Physical Science Honors students had remarkable success in growing protein crystals, with 61% of students observing crystals in at least one of their Limbro plate wells. The majority of groups with crystals observed large protein crystals in wells with intermediate salt concentration (0.7 – 1.3M), and several groups observed many smaller, presumably salt crystals in 1.3 -1.5M NaCl. Physical Science Honors students answered on average 2.4 of the objective questions correctly. Most frequently missed were the questions 2 and 3, about the physical chemistry of protein crystallization.

The Biology Honors students were most successful at growing crystals (90% of groups observed crystals). Many factors may have contributed to their success, however the most obvious was that were co-enrolled in Biotechnology 1 and thus had more experience using micropipettes. They were able to set-up all 24 of the salt/pH/drop conditions specified in the McKenna protocol. These students also had more laboratory experience in general (the Biotechnology 1 class had recently completed a PCR lab), and that may have also contributed to their confidence in following complex instructions. This group of students answered an average of 3.5 of the objective questions correctly. They most often missed question 3 which asked students to explain differences in crystal formation under the different salt concentrations.

The summary data from the “attitudes towards biotechnology” survey are presented in Table 1. Because the Physical Science Honors group included 81 students, and the Biology Honors group included only 21 students, differences may not be significant. Interesting differences between the two groups included that more of the Biology Honors indicated their problem solving skills increased as a result of the activity, and more of the Physical Science Honors students indicated an increased interest in biotechnology as a result of the activity. Physical Science Honors indicated a greater interest in pursuing a career in biotechnology, though the Biology Honors had previously self-selected to enroll in a biotechnology course. Both groups equally enjoyed the activity. On average, neither group was in favor of using tax dollars to support biotechnology research!

Literature cited:

Dawson, V. (2007). An Exploration of High School (12–17 Year Old) Students’ Understandings of and Attitudes Towards Biotechnology Processes. Research in Science Education 37, 59–73. Dawson, V., & Soames, C. (2006). The effect of biotechnology education on Australian high school students' understandings and attitudes about biotechnology processes. Research in Science & Technological Education, 24: (2), 183-198. Dunham, T., Wells, J., & White, K. (2002). Biotechnology Education: A Multiple Instructional Strategies Approach. Journal of Technology Education, 14 (1) 65-81.  Budget and budget justification:

Most of the materials purchased for this project were generously supplied by the CPET group at the University of Florida.

Table 1. Average responses by Physical Science Honors and Biology Honors students to a survey about their attitude towards a protein crystallization lab and biotechnology.

Average Response

(1= strongly disagree, 2= disagree, 3=undecided, 4= agree, 5 = strongly agree

Physical Science Honors Biology Honors

1. I feel good about my problem solving skills. 4.43 4.30

2. My problem solving skills increased as a result of this laboratory activity 2.77 3.30

3. My understanding of biotechnology has increased as a result of this activity. 3.35 3.05

4. My interest in biotechnology has increased as a result of this activity. 4.05 3.15

5. I enjoyed the experimental aspects of this activity. 4.00 4.00

6. I want to learn more about jobs in the biotechnology industry. 2.62 2.85

7. I am interested in pursuing a career in biotechnology. 3.63 2.90

8. I have an increased understanding of the processes associated with “rational drug design”. 2.57 2.70

9. I have an increased understanding of the ways that mathematics can be used in biology. 2.47 3.40

10. Research on medical applications of biotechnology should be funded by tax dollars. 2.97 2.90

Modifications from original proposal:

Because Physical Science students were unsuccessful in both the micropipetting and protein crystallization labs, data pertaining to their interests and understanding of biotechnology was not collected or analyzed. These students are a difficult group to teach because they generally do not listen well and do not follow instructions. In addition, they are scheduled for the last period of the day, when they are generally restless and their attention spans are shortest. The group of Biology Honors students were included in this activity because they were participating in the Biotechnology Academy, and because they expressed a desire to perform the lab when they saw it setup for the physical science classes. Objective multiple choice pre- and post- tests were not administered to the students because they were all enrolled in different courses that actively working on different aspects of their course’s required content. I originally thought it would be possible to pace the Physical Science and Physical Science Honors students the same, but major differences in the text book organizations and student populations prevented that from happening. Insights: There are significant cognitive and behavioral differences between groups of ninth grade high school students that are enrolled in Physical Science, Physical Science Honors, and Biology Honors. If I were to do this again, I would not attempt it with Physical Science students. The instructions were too complex for them and it was not realistic for them to complete the activity during a 45 minute class period. Honestly I did not learn much from the formal action research process. It consumed a large amount of my time that could have been better spent developing and planning activities for my students. The formality seemed superimposed and did not provide better insight than I would have gotten had I done what I normally do, which is plan lessons, execute them, reflect upon them both by myself and with my colleagues, and then modify them. I have attached my reflective journal at the end of this document. Dissenimation:

I shared information from the CPET Bench to Bedside Summer Institute at the University of Florida with two colleagues in my school, one who teaches Biotechnology 1, 2, and 3, and one who teaches Biology Honors and AP Biology. The biotechnology colleague will attend the summer institute this coming summer. I will continue to share the insights I gained into medical application of biotechnology with my students whenever and wherever they are appropriate.

Reflections on Action Research Activity: 

Sunday April 10, 2011 – I re‐read all the protein crystallization handouts from B2B, modified the powerpoint from the Tartaglia/McKenna lab, and color‐coded the instruction sheet to try and make the instructions more user friendly for my students. 

Monday April 11, 2011 

I’ve stayed late and made copies of the activity, made the Na‐Acetate, Tris, and NaCl, and aliquoted the solutions into 50 tubes that I color coded with tape.  I’ve also aliquoted ddH2O.  This alone has taken nearly 2 hours.   

During class I gave a shortened version of the Tartaglia powerpoint on Protein Crystallization to 2 classes: Periods 2 and 3 for Honors Physical Science.  My goal was to introduce the concept of proteins and their importance in biology, and to explain why scientists want to know their structure so they can design drugs to modify their activity.  I think most students “got it”. 

Tuesday April 12, 2011 

Today I used the powerpoint to explain the Protein Crystallization activity to periods 4,5, and 7.  7th period is my lone class of “standard” Physical Science – and as always, they were the least attentive.  However, probably due to the morning FCAT testing, 1/3 of the students were absent.  Unfortunately, those students didn’t get the introduction/justification.  Periods 4 and 5 had no absentees.  They’re my Honors Physical Science Students. 

I’m setting up the lab for teaching micropipetting – using the colored dyes.  I’m making 10 trays with the 96 well plates, two colored waters, and instructions.  I need to pick up the pipetters from Dr. Kehoe after 1st period, return them for her 6th period, and then get them  again for my 7th period. 

Wednesday April 13, 2011 

I gave a 10 minute introduction to the lab, explaining the different sizes of pipettes, how to adjust volumes, how to located wells on a 96‐well plate, etc.  One of Dr. Kehoe’s seniors helped teach micropipetting to the 2nd period class.  The activity went pretty well.  In hindsight, this class worked best, even though I did a better job of explaining the activity as the day went on.  I think they talked to their friends during the day, because as the day went on, students seemed to want to skip the pipetting of the larger volumes once the identified the pattern.  Really every class did an excellent job.  Our Wednesday classes are only 39 minutes long, so it was a full period, but they finished and seemed satisfied with their results.  Some of the students took pictures of their completed dishes with their i‐Phones.  There was a sense of pride.   

The only class that had multiple mistakes was my 7th period standard PS class: nearly every group messed up in one way or another (letters shifted, volumes not the same, spilled entire dish, in one case dumped entire dish after one student noted that you can invert the dishes and the liquid stays in).   

I spent about 2 hours after school prepping the protein crystallization lab – setting trays with all the required solutions, cover slips, forceps, immersion oil in bottles (these are cute), limbro trays, micro bridges, and pipette tips.  I’ll have to borrow the pipettes from Dr. Kehoe tomorrow – picking them up after 1st period, returning after 5th, and picking up again after 6th. 

Thursday April 14, 2011 

First I got ice, then I reconstituted the lysozyme in 400 ul buffer.  I did the lab with 5 classes: 2‐5 are Physical science Honors students and 7th period is physical science standard. 

2nd period  ‐ After describing the lab, and writing the color coding for the solutions and the “cheats” for the pipetters on the white board, the students did their best.  Only one group set up all 24 wells and added the hanging drops and the sitting drops.  Most of the other groups have their solutions made, but haven’t added protein.   I had them cover the trays and write their names on the cover.  We’ll have to complete this Friday.   

3rd period – I decided to reduce the number of wells each group set up in half, and divided the room into 2 groups – 6 with sodium acetate buffer and 4 with Tris.  This class worked very diligently and every group set up 12 wells.  The timing was MUCH better than with 2nd period. 

I spend lunch replenishing solutions and cover slips and bridges. 

4th Period – these students didn’t do so well – only 1/3 finished the set up of 12 wells.   This class meets right after lunch and these students were pretty distracted.  Also – today was Spring Art Festival (announced today!), and at lunch there were bands.   

5th Period – About the same as 4th period, except because I have 6th period planning, I finished the set up for a couple of groups.  Many were nearly finished and seemed invested enough to want to complete and be late to their next class. 

Overall – with the Honors students, the group skills and problem solving were nice to overhear.  Many weren’t confident with setting volumes on the pipetters, and they dialogued productively.  Good group work.    Most students were focused and gave the lab their best shot.    

7th Period – No group finished the activity.  They lacked confidence in their lab skills, socialized, asked to go to the bathroom.  One group just added some liquids to each well, and at first I was impressed at how quickly they were working.  However, when they added the bridges and they were underwater, I caught on that they were just adding liquids randomly and not attempting to measure volumes correctly.  They were the “worst”. 

Though I don’t expect to see many crystals next week, and I am EXHAUSTED, no one got hurt, no one broke any expensive equipment, and I  believe the students got a glimpse into the world of biotechnology lab work.  I’m going to let my Biology Honors students do the lab tomorrow (many are in the biotechnology academy and have good pipetting skills). 

April 20, 2011 – Yesterday I peaked at one groups Limbro dishes from PSH and there were large, beautiful crystals in some of the drops.  Today I set up dissecting microscopes, instructed students in their proper use, and let them search for crystals. All but two groups in the Bio Hon class had crystals, all but one group in PSH second period had crystals (some had salt crystals at the high salt concentrations, but most had nice large protein crystals at the mid range of salt, especially in Na‐acetate buffer (pH 4.2), and overall, 61% of the students grew crystals in PSH.  The students were definitely pumped up when they found crystals, and worked enthusiastically to find and sketch them.  They even took pictures with their iphones.  It was fun watching and listening to them. 

The not‐ so‐ unexpected bad news – only 1 group out of 10 in Standard Physical Science Class grew crystals.  I am not going to continue the analysis of that group because I don’t think there’s any point in it.  They didn’t follow instructions, they didn’t understand what they were doing, and I need to help them review for a quiz on chemical bonding. 

April 21,2011 – Here’s a down side – I had to wash 70 Limbro dishes today – it took over an hour.  I just can’t bring myself to throw away that high quality plastic.  Though I doubt I’ll ever grow crystals with students again, I think these dishes may have use in catalase labs or even population growth studies with yeasts!  So ‐  I washed them. 

April 22, 2011 – The final report is due today, so I’m working away analyzing data.