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RET Teaching RET Teaching ModuleModule

Bill Farmer – Evanston Township High SchoolBill Farmer – Evanston Township High SchoolUniversity of Illinois, ChicagoUniversity of Illinois, Chicago

Advisor: Prof. David SchneeweisAdvisor: Prof. David SchneeweisGraduate Student Mentor: Sujata Sundara-RajanGraduate Student Mentor: Sujata Sundara-Rajan

REU Students: Amin Farokhrani and Jaime McCoin REU Students: Amin Farokhrani and Jaime McCoin Summer 2005Summer 2005

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Presentation OutlinePresentation Outline

RET BackgroundRET Background Teaching Module OverviewTeaching Module Overview Data AnalysisData Analysis Inquiry LabInquiry Lab Outcomes and StandardsOutcomes and Standards

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RET BackgroundRET Background The primary objective of my research group was the The primary objective of my research group was the

development of an interface between a small biocompatible development of an interface between a small biocompatible microchip and an existing biological system. microchip and an existing biological system.

Such technology would be applicable to a design for a retinal Such technology would be applicable to a design for a retinal prosthesis to restore partial vision to patients who have prosthesis to restore partial vision to patients who have experienced photoreceptor degeneration.experienced photoreceptor degeneration.

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RET BackgroundRET Background Creating such an interface requires the Creating such an interface requires the

development of an effective and reliable development of an effective and reliable method to guide the growth of human timethod to guide the growth of human tissue cells, primarily retinal neurons, in ssue cells, primarily retinal neurons, in such a way so that it establishes a connesuch a way so that it establishes a connection between the body and the microchction between the body and the microchip.ip.

Our research group’s end goal was to cOur research group’s end goal was to conduct in vitro tests of a soft lithographonduct in vitro tests of a soft lithography micropatterning technique with humay micropatterning technique with human cell tissue cultures.n cell tissue cultures.

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My ContributionMy Contribution In preparation for the in vitro tests I wIn preparation for the in vitro tests I w

as responsible for the growth and maias responsible for the growth and maintenance of human fibroblast cells.ntenance of human fibroblast cells.

Human fibroblast cells were used initiHuman fibroblast cells were used initially due to their relative ease in cell culally due to their relative ease in cell culture compared to retinal neuron cells. ture compared to retinal neuron cells.

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Fibroblast Tissue Fibroblast Tissue CulturingCulturing

Utilization of sterile tecUtilization of sterile techniquehnique

Cells maintained at Cells maintained at 37°C and 5% carbon 37°C and 5% carbon dioxidedioxide

Passage of cells at Passage of cells at 75% confluence75% confluence

10x Magnification Fibroblast Cells

20x Magnification Fibroblast Cells

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Data Acquisition For My Data Acquisition For My ClassroomClassroom

Digital images were acquired to track Digital images were acquired to track the growth patterns of the fibroblast the growth patterns of the fibroblast cell cultures. cell cultures.

I set up a controlled experiment I set up a controlled experiment where I manipulated the initial where I manipulated the initial concentration of cells and monitored concentration of cells and monitored its impact on growth rates. its impact on growth rates.

Digital images of fixed cells stained Digital images of fixed cells stained with DAPI were also taken for with DAPI were also taken for potential use in the classroom. potential use in the classroom.

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Teaching Module Teaching Module OverviewOverview In order to incorporate my RET experience In order to incorporate my RET experience

into my classroom I designed a two part into my classroom I designed a two part mini unit for my mixed-level biology mini unit for my mixed-level biology classes. classes.

The first part of the unit involves the The first part of the unit involves the student analysis of the digital fibroblast cell student analysis of the digital fibroblast cell data.data.

The second part of the unit involves the The second part of the unit involves the development of an inquiry-based lab where development of an inquiry-based lab where students will design their own controlled students will design their own controlled experiment to test the impact of a chosen experiment to test the impact of a chosen variable on the growth rate of bacteria.variable on the growth rate of bacteria.

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Analysis of Fibroblast Analysis of Fibroblast DataData Students will…Students will…

Utilize freeware Scion Image software Utilize freeware Scion Image software

available through the NIH website to view available through the NIH website to view the images of the human fibroblast cells the images of the human fibroblast cells calculate the cell population and densities. calculate the cell population and densities.

Organize their data in computer generated Organize their data in computer generated tables and graphs using Microsoft Excel.tables and graphs using Microsoft Excel.

Analyze the data and draw conclusions with Analyze the data and draw conclusions with regard to the impact of initial cell regard to the impact of initial cell concentration on the growth rate of human concentration on the growth rate of human fibroblast cells.fibroblast cells.

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Analysis of Fibroblast Analysis of Fibroblast DataData Sample Excel Graph of Fibroblast DataSample Excel Graph of Fibroblast Data

Fibroblast Cell Growth at Different Initial Volumes of a Known Concentration of Cells

0

20

40

60

80

100

120

140

160

180

1 2 3 4

Time (Days)

Po

pu

lati

on

of

Cel

ls

25 Microliters

50 Microliters

75 Microliters

100 Microliters

125 Microliters

150 Microliters

200 Microliters

500 Microliters

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Analysis of Fibroblast Analysis of Fibroblast DataData This portion of the module will be conThis portion of the module will be con

ducted through the course of two 43 mducted through the course of two 43 minute class periods. inute class periods.

The only materials necessary for this The only materials necessary for this module are computers available either module are computers available either in the computer center or class sets of in the computer center or class sets of portable laptop go-carts. portable laptop go-carts.

Students will generate a typed formal lStudents will generate a typed formal lab report.ab report.

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Inquiry Lab Inquiry Lab Students will… Students will… Develop a procedure for a controlled Develop a procedure for a controlled

experiment to test how the manipulation experiment to test how the manipulation of their chosen variable impacts the rate of their chosen variable impacts the rate of bacterial growth.of bacterial growth.

Learn about sterile technique and how to Learn about sterile technique and how to culture bacteria cells. culture bacteria cells.

Conduct their designed experiments and Conduct their designed experiments and gather data on bacterial growth patterns. gather data on bacterial growth patterns.

Organize and analyze their data in a Organize and analyze their data in a formal lab report write-up. formal lab report write-up.

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Inquiry Lab Inquiry Lab This portion of the module will be This portion of the module will be

conducted through the course of 4 days. conducted through the course of 4 days. This includes two 43 minute single periods This includes two 43 minute single periods and two 86 minute double periods. and two 86 minute double periods.

Two used incubators have been acquired Two used incubators have been acquired from other teachers within my department from other teachers within my department which will enable my class to culture which will enable my class to culture bacteria. Learn about sterile technique and bacteria. Learn about sterile technique and how to culture bacteria cells. how to culture bacteria cells.

The remaining equipment necessary for The remaining equipment necessary for culturing bacteria is available within the culturing bacteria is available within the school’s supplies.school’s supplies.

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ETHS Standards ETHS Standards 19. Formulate testable hypotheses.19. Formulate testable hypotheses. 20. Design and conduct scientific investigations 20. Design and conduct scientific investigations

by identifying and clarifying the question, by identifying and clarifying the question, method, controls and variables; organizing and method, controls and variables; organizing and displaying data; and explaining the results.displaying data; and explaining the results.

21. Know scientific explanations or models 21. Know scientific explanations or models must be: logically consistent supported by must be: logically consistent supported by evidence, open to questions and possible evidence, open to questions and possible modifications, and based on historical and modifications, and based on historical and current scientific knowledge.current scientific knowledge.

22. Use technology and math to improve 22. Use technology and math to improve investigations and communications; use investigations and communications; use computers for the collection, analysis, and computers for the collection, analysis, and display of data. display of data.

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Illinois Science Illinois Science Standards Standards 11.A.4a 11.A.4a Formulate hypotheses referencing Formulate hypotheses referencing

prior research and knowledge.prior research and knowledge. 11.A.4b Conduct controlled experiments or 11.A.4b Conduct controlled experiments or

simulations to test hypotheses. simulations to test hypotheses. 11.A.4c Collect, organize and analyze data 11.A.4c Collect, organize and analyze data

accurately and precisely. accurately and precisely. 11.A.4d 11.A.4d Apply statistical methods to the Apply statistical methods to the

data to reach and support conclusions. data to reach and support conclusions. 11.A.4e Formulate alternative hypotheses 11.A.4e Formulate alternative hypotheses

to explain unexpected results. to explain unexpected results. 11.A.4f Using available technology, report, 11.A.4f Using available technology, report,

display and defend to an audience display and defend to an audience conclusions drawn from investigations. conclusions drawn from investigations.

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Questions?Questions?Questions WelcomeQuestions Welcome

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AcknowledgementsAcknowledgements National Science FoundationNational Science Foundation University of Illinois, ChicagoUniversity of Illinois, Chicago Prof. Andreas Linninger - DirectorProf. Andreas Linninger - Director Prof. Christos Takoudis – Co-DirectorProf. Christos Takoudis – Co-Director Prof. David Schneeweis - AdvisorProf. David Schneeweis - Advisor Amin Farokhrani and Jaime McCoin – REU StudentsAmin Farokhrani and Jaime McCoin – REU Students

NSF RET Fellowship at UIC

ReferencesReferences 1. H Kolb. How the retina works. American Scientist, Jan.-Feb. 2003.1. H Kolb. How the retina works. American Scientist, Jan.-Feb. 2003. 2. J. Chang et al. A modified microstamping technique enhances polylys2. J. Chang et al. A modified microstamping technique enhances polylys

ine transfer and neuronal cell patterning. Journal of Biomaterials, 2003.ine transfer and neuronal cell patterning. Journal of Biomaterials, 2003. 3. D. Branch et al. Long-term stability of grafted polyethylene glycol surg3. D. Branch et al. Long-term stability of grafted polyethylene glycol surg

aces for use with microstamped substrates in neuronal cell culture. Jouaces for use with microstamped substrates in neuronal cell culture. Journal of Bimaterials, 2001.rnal of Bimaterials, 2001.

4. D. Branch et al. Long-term maintenance of patterns of hippocampal4. D. Branch et al. Long-term maintenance of patterns of hippocampal pyramidal cells on substrates of polyethylene glycol and microstamped pyramidal cells on substrates of polyethylene glycol and microstamped

polylysine. IEEE Transactions on Biomedical Engineering, March 2000.polylysine. IEEE Transactions on Biomedical Engineering, March 2000. 5. A. Bernard et al. Microcontact printing of proteins. Journal of Advanc5. A. Bernard et al. Microcontact printing of proteins. Journal of Advanc

ed Materials, July 2000.ed Materials, July 2000. 6. B.Wheeler et al. Microcontact printing for precise control of nerve ce6. B.Wheeler et al. Microcontact printing for precise control of nerve ce

ll growth in culture. Journal of Biomechanical Engineering, February 19ll growth in culture. Journal of Biomechanical Engineering, February 1999.99.

7. R. Kane et. al. Patterning proteins and cells using soft lithography. Jo7. R. Kane et. al. Patterning proteins and cells using soft lithography. Journal of Biomaterials, 1999.urnal of Biomaterials, 1999.

8. D. Branch et al. Microstamp patterns of biomolecules for high-resolut8. D. Branch et al. Microstamp patterns of biomolecules for high-resolution neuronal networks. Medical & Biological Engineering & Computing, ion neuronal networks. Medical & Biological Engineering & Computing, 1998.1998.

9. B. Ujhelyi and K. Garsha, Patterned growth of neurons in vitro. Beck9. B. Ujhelyi and K. Garsha, Patterned growth of neurons in vitro. Beckman Institute for Advanced Science and Technology, UIUC, 2003.man Institute for Advanced Science and Technology, UIUC, 2003.

10. B. Wheeler et al., 10. B. Wheeler et al., http://soma.http://soma.npanpa..uiucuiuc..eduedu/labs/wheeler/home.html/labs/wheeler/home.html. UIUC, 2005.. UIUC, 2005.