Integrating research and advanced microscopy in the high school curriculumCraig Queenan, C.E.M.T. & David BeckerBergen County Academies, Nano-Structural Imaging LabSPIE Defense, Security, & Sensing 2012Conference 8378: Special Session on Microscopy for STEM EducatorsApril 26, 2012

Voices for Change“Every country on earth at the moment is reforming public education… People are trying to work out, ‘How do we educate our children to take their place in the economies of the 21st century’.”1

-Sir Ken RobinsonInternational leader in education development

“The battle for America's future will be won or lost in the next century in America's institutions of public education…because they are the crucible in which the young people who will comprise the largest part of our population a quarter century from now will be formed.”2

-Lawrence H. Summers Former Director of the White House US National Economic Council

President Emeritus of Harvard University

Today’s Goals• STEM education in the U.S.

• National STEM initiatives

• BCA’s position within the STEM community

• Program perspectives• Integrating research and microscopy

STEM in the U.S.

“There is evidence that top U.S. students, who have disproportionate potential to become future innovators, are eschewing careers in S&E.”4

“Today in the U.S., just 16 percent of graduates receive degrees in STEM fields, compared to China, where 52 percent of graduates earn degrees in these critical fields”5

3

Global Trends - Education

U.S. – little change in number of STEM bachelor degrees

China – large increase in number of STEM bachelor degrees

U.S. and China both showing increases in number of doctoral degrees, with China’s

trend surpassing U.S.

U.S. degrees increasingly from foreign born students rather than citizens

I

4 4

National STEM InitiativesThere are a number of groups that are actively promoting changes to STEM education:

The National Science Board (part of NSF) -May 2010 report “Preparing the Next Generation of STEM Innovators”6

Called for an elevation of the ceiling of achievement for future innovators through the expansion of: •inquiry-based learning •peer collaboration•open-ended real-world problem solving •hands-on training•interactions with practicing scientists, engineers and other experts

National STEM InitiativesPresident Obama’s 2009 “Educate to Innovate” initiative7

•The program seeks to “apply new and creative methods of generating and maintaining student interest and enthusiasm in science and math, reinvigorating the pipeline of ingenuity and innovation essential to America’s success that has long been at the core of American economic leadership.”

• ARPA-ED

The STEM Education Coalition’s 2009 “Letter to POTUS”8

•Signed by over 200 companies, universities, institutes and organizations representing over 6.2 million STEM professionals and educators

• i.e.: INTEL, 3M, ACS, SPIE, Carnegie Mellon University

•Pledge to engage members to form partnerships with educators, inspire students to participate in STEM, improve hands-on lab environments for students, raise public awareness.

• 10,000 communities of support • National STEM Week • National Lab Day

National STEM InitiativesThe National Research Council (of the National Academies)9

•Framework for science education•Contains eight practices which they distinguish from skills in that the practices, “stress that engaging in scientific inquiry requires coordination both of knowledge and skill simultaneously.”

9

STEM in the U.S.The initiatives and programs set forth by these groups do an excellent job in improving STEM outreach and education to key underrepresented demographics in our society:

•Women

•Minorities

•Children of low-income families

The Bergen County Academies•Fostering and Cultivating STEM Leadership

10

Research & Microscopy at BCAResearch supplemental to the STEM education students receive and reinforces traditional classroom teaching

A Journey to the Frontiers of Science•Students emphasis on:

• Understanding what tools, efforts, accomplishments and failures have led researchers to current boundaries of research

• Developing a novel research project (with faculty guidance)• Gathering a working understanding of the tools, techniques and skills

they will need to master through hands-on training and practice• Collecting data and analyzing results independently and amongst peers• Presenting findings to an audience• Science competition or fair; Poster presentation; Publication

Student Skill AttainmentNRC’s framework is still in its early stages and not adopted

•BCA’s STEM education rests upon 6 pillars, or transferable skill sets that can be carried into college, graduate school, and careers.• Context and Perspective• Tools and Techniques• Collaboration• Analysis and Justification• Quality and Failure• Defense and Presentation

The confidence in these skills that a student is able to craft gives them an advantage in future endeavors, and puts them on course to become part of the next generation or STEM Leaders!

Success & Accolades• College enrollment• Academy for the Advancement of Science 2012

• 72 students

• 39% (28 of 72) enrolled in an Ivy League schools or in a U.S. News top 10 Engineering School in the U.S. 11 for next fall12:• Carnegie Mellon University – 2• Columbia University – 5• Cornell University – 5• Harvard University – 1• Massachusetts Institute of Technology – 1• Princeton University – 6• University of California, Berkeley – 6• Yale University - 2

Success & Accolades• Intel Science Talent Search• 7 semi-finalists and 2

finalists since 2009

• Publications• 16 abstracts published by 18

students in the Proceedings of Microscopy and Microanalysis since 2008

• 4 of those students attended the national conference and presented their posters

X-ray Microanalysis as a Tool for Analyzing Stem Cell DifferentiationLeora Aizman*, Danielle Hantman*, Craig Queenan*, Alyssa Calabro*,

David Becker*, Robert Pergolizzi*, and Marika Bergenstock*** Bergen County Academies, Nano-Structural Imaging Lab, 200 Hackensack Avenue, Hackensack, NJ 07601

** 3D Biotek, LLC., Technology Center of New Jersey, 675 US Highway One, North Brunswick, NJ 08902

IntroductionCell culture serves as an important tissue model in research

labs, allowing the effects of a treatment to be examined in a reproducible, relatively inexpensive controlled environment. Although 2D cell culture is most common, it does have drawbacks including limited cell-cell interactions and a possible disconnect between cellular behavior in vivo and in vitro. One method that is gaining popularity in response to these issues is culturing cells on a 3D matrix, or scaffold. The polycaprolactone (PCL) scaffolds available through 3D Biotekare non-toxic, have well-defined pore size and fiber diameter and are free of animal-derived material. In addition, the PCL scaffold is biodegradable, meaning that the scaffold can be introduced into an in vivo system from an in vitro system to examine the true effects on an organism. This study involves the use of scanning electron microscopy (SEM) and energy dispersive X-ray spectroscopy (EDS) to analyze the differentiation of human mesenchymal stem cells (hMSC) into osteoblasts on PCL scaffolds. Differentiation would be indicated by the presence of osteoblast nodules composed of calcium phosphate, a main component of bone tissue.

Acknowledgements•Dr. Howard Lerner, Superintendent, Bergen County Technical Schools & Special Services• Edmund Hayward, Technology Director, Bergen County Technical Schools• Russell Davis, Principal, Bergen County Academies• The research team at 3D Biotek for their collaboration on this project

Reference[1] MK Bergenstock, et al., Novel Fully Biodegradable Biomimetic Scaffolds for Bone Regeneration and Repair. Poster session presented at: Society for Biomaterials 2011 Annual Meeting and Exposition; April 13-16, 2011; Orlando, FL.

Materials & MethodsCell Seeding onto Scaffolds [1]:

An aliquot of either human dermal fibroblast cells (hFB) or hMSCs in suspension was pipetted onto the center of each PCL scaffold and incubated for 3 hours at 5% CO2 and 37 � C to allow cells time to infiltrate prior to the addition of growth media. Twenty-four hours after seeding, the hMSCs’ growth media was replaced by osteoblastic differentiation media. The hFB growth media and osteoblastic differentiation media were replenished every 48 hours. The scaffolds were then placed in SEM fixative at the time points: 3 weeks, 4 weeks, and 5 weeks of growth.

SEM Sample Preparation:At the desired time points, scaffolds were placed in SEM

fixative (5% glutaraldehyde in PBS). The scaffolds were post-fixed with 2% osmium tetroxide, dehydrated in a graded series of ethanol, critical point dried, mounted on aluminum pins and coated with carbon. Samples were imaged with an FEI Quanta 200 3D and both quantitative analysis as well as elemental mapping were performed using the Oxford INCA EDS system.

Results & DiscussionAn unseeded, control scaffold was imaged to understand

its inherent structure and texture, and EDS analysis was performed to establish a baseline elemental background. The scaffold appeared to have a porous surface and was smooth (Figure 1B). The only elements present in the empty scaffold were carbon and oxygen (Figure 1A), as was expected; thus the presence of calcium and phosphorus in any seeded sample would be a result of osteoblastic differentiation.

After 3 weeks of growth, both hFBs and hMSCs were visible on the respective scaffolds. Fibroblast cells appear closely adhered to the scaffold (Figure 2A), while hMSCs were beginning to spread across the fibers of the scaffold (Figure 2 B&C). EDS analysis of the hFB scaffold, similar to the control, showed an absence of calcium and phosphorous, as was expected (Figure 2A). The quantitative EDS analysis of the hMSC scaffold showed the presence of calcium and phosphorous (Figure 2B), indicating osteoblasticdifferentiation, and X-ray mapping showed these elements to be localized in areas of apparent cell growth (Figure 2C).

After 4 weeks of development, the hFB and hMSC cells continued to spread across the scaffold fibers. In comparison to the textured appearance of the hMSCs (Figure 3B), the fibroblast cells grew in a smoother and flatter form (Figure 3A). EDS analysis of the hFB scaffold showed no presence of calcium or phosphorous (Figure 3A). Elemental mapping of the hMSC scaffold continued to show the presence of calcium and phosphorous in corresponding areas of cell growth (Figure 3C).

After 5 weeks of growth, hFBs grew evenly across the scaffold (Figure 4A), whereas the hMSCs continued to grow in clusters, remaining close to the fibers (Figure 4B). EDS analysis showed the presence of calcium or phosphorous only on the hMSC scaffold (Figures 4A&B). Elemental mapping of the hMSC scaffold shows calcium and phosphorus localized in the same areas suggesting the presence of calcium phosphate nodules that are characteristic of osteoblasts (Figure 4C).

In this study, cells were successfully grown and differentiated on the 3-dimentional scaffolds. Overall, there appeared to be an increasing trend in the amount of calcium and phosphorous detected among hMSC scaffolds indicating the continued differentiation of hMSCs into osteoblasts. The data presented here shows that energy dispersive X-ray spectroscopy is a useful tool for the identification and analysis of differentiated stem cells with unique elemental markers.

Figure 1: Unseeded control scaffold. (A) SEM micrograph and quantitative EDS analysis. Note the presence of only carbon and oxygen. (B) Higher magnification micrographs of control scaffold.

Figure 2: Seeded scaffolds at 3 weeks growth. (A) SEM micrograph and quantitative EDS analysis of hFb cells. (B) SEM micrograph and quantitative EDS analysis of hMSC cells. Note the presence of calcium and phosphorous. (C) SEM micrograph with EDS maps showing localization of calcium and phosphorous.

Figure 3: Seeded scaffolds at 4 weeks growth. (A) SEM micrograph and quantitative EDS analysis of hFb cells. (B) SEM micrograph and quantitative EDS analysis of hMSC cells. Note the presence of calcium and phosphorous. (C) SEM micrograph with EDS maps showing localization of calcium and phosphorous.

Figure 4: Seeded scaffolds at 5 weeks growth. (A) SEM micrograph and quantitative EDS analysis of hFb cells. (B) SEM micrograph and quantitative EDS analysis of hMSC cells. Note the presence of calcium and phosphorous. (C) SEM micrograph with EDS maps showing localization of calcium and phosphorous.

Presented at Microscopy & Microanalysis 2011August 7-11, Nashville, TN

Poster Number: 219 Paper Number: 81466

The Bergen County Academies

• Bergen County Academies (BCA), Hackensack, NJ• 1952 - Opened as Bergen County Vocational-

Technical High School• 1980’s - Began to emphasize computers,

physics, electronics• 1992 - Became BCA with the Academy for the

Advancement of Science and Technology• Currently 6 additional Academies :

• Engineering & Design Technology• Medical Science Technology• Telecommunications & Computer Science • Business & Finance• Culinary Arts & Hotel Administration • Visual & Performing Arts

BCA Info • Serves residents of Bergen County New

Jersey• 72 towns; nearly one million residents

• Magnet high school• Draws from the best prepared and most

academically invested rising ninth graders

• Tuition free for students and families• Free of economic barriers that impede

STEM education

• Student body• ~1,100 students in seven career focused

academies

• Extended day (8:00AM-4:10PM)• Students generally carry 12-15 courses

each trimester• More opportunity to take specialized

courses and electives

Core STEM education at BCA

Required courses for all students in the Science Academy at BCA

Research at BCA“Learn Science By Doing Science”•Dedicated faculty (many coming from industry)•Provided the tools needed for high-quality research

• September 2004 – Cell Biology Lab (Biotech/Cell Biology)• September 2006 - Stem Cell Lab (Molecular Biology/Genetics) • May 2008 - Nano-Structural Imaging Lab (Microscopy)• September 2009 - Nanotechnology Lab (Chemistry)

• Funding Sources:•NJ DOE Carl D. Perkins Vocational and Technical Grant to support the initial facility development of the research labs•County Bond funding from the Bergen County Board of Chosen Freeholders•BCTS District Technology Department budget

Research at BCA• Independent research • Open to students from any of the seven academies

• Not only science and engineering students

• Students treated more like graduate students than high school students• Taught to:

• Search the primary literature for a topic and develop a novel project• Find the materials that are needed and order those that are not already available at the

school• Be responsible for maintenance of project (i.e. cell culture) as well as time management• Determine statistical significance in their data• Write, present and defend their work

• All skills that will benefit the students in college and careers

Nano-Structural Imaging Lab• Core imaging facility featuring SEM, TEM, LSCM and

support/sample preparation equipment• Self-sufficient facility• Staffed and operated independently of academic departments• Research using microscopy:• Independent research

projects from other labs

• Independent research projects from NSIL

• Collaborative projects

• Internships

• Summer research program

• Group exercises

NSIL - How It’s Used

*Student figures from July 1, 2010 – June 30, 2011

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5

17238

12

260

Microscopy Integration into Curriculum• Introduction to Microscopy Elective• Open to all students – pre-requisite for microscopy research• Subject matter has foundations in Biology, Chemistry, Physics,

Engineering and History• Students learn:

• Microscopy Theory• History, principles and discoveries, structure and design of LM, SEM, and TEM, image

formation

• SEM and TEM• Sample preparation, safety, proper use and maintenance, quality data

• All lecture supplemented with hands-on exercises and lessons.

Microscopy Integration into Curriculum• Enhanced Classroom Experiences• Ex. Biology classes learning about the parts of a cell

• Students use the TEM to identify and image organelles• Reinforces classroom learning; excites students

• “Spark” Events• Non-science classes• Visual arts and graphic design class – Art in science

• Students learn about the SEM and some of the principles behind imaging• Image samples that they collected and brought to the lab• Use images in a lesson on introducing them to Adobe Photoshop colorization

Successes of Research at BCA• Early exposure to technology and techniques• Confidence in research skills built upon the 6 “pillars”• Outreach to students outside of the sciences• Science fair and competition success • Collaboration with industry and academic professionals• College admissions and program enrollment• Publication and conference presentation

Future• Tracking of students post college and into careers

• Increased focus on the three areas deemed necessary in the development of future leaders:• Research, entrepreneurship, and global studies

References1. Robinson, K., “RSA Lecture - Changing Paradigms,” Royal Society for the encouragement of Arts, Manufactures and Commerce,

14 October 2010. <http://www.thersa.org/events/video/archive/sir-ken-robinson> (16 April 2012).2. Summers, L., “Remarks of President Lawrence H. Summers, ‘Learning With Excitement' Conference,” Harvard Graduate School

of Education, 03 October 2003. <http://www.hks.harvard.edu/fs/lsummer/speeches/2003/afterschool.html> (16 April 2012).3. Lowell, B., et al., “Steady as she goes? Three generations of students through the science and engineering pipeline,” Rutgers

University, October 2009. <http://policy.rutgers.edu/faculty/salzman/steadyasshegoes.pdf> (16 April 2012).4. “Science and Engineering Indicators 2010,” National Science Board of the National Science Foundation, January 2010.

<http://www.nsf.gov/statistics/seind10/start.htm> (16 April 2012).5. Hook, L., “U.S. falling behind in supplying science, technology, engineering and math degrees,” The Washington Times, LLC, 23

March 2012. <http://www.washingtontimes.com/news/2012/mar/23/help-wanted-855590724/> (16 April 2012).6. “Preparing the Next Generation of STEM Innovators: Identifying and Developing Our Nation’s Human Capital,” National

Science Board Committee on Education and Human Resources, 5 May 2010. <www.nsf.gov/nsb/publications/2010/nsb1033.pdf> (16 April 2012).

7. “President Obama Launches ‘Educate to Innovate’ Campaign for Excellence in Science, Technology, Engineering & Math (STEM) Education,” White House, Office of the Press Secretary, 23 November 2009. <http://www.whitehouse.gov/the-press-office/president-obama-launches-educate-innovate-campaign-excellence-science-technology-en> (16 April 2012).

8. “Letter to the President of the United States,” STEM Education Coalition, 20 November 2009. <http://www.nationallabnetwork.org/pdfs/letter_to_POTUS_Nov20.pdf> (16 April 2012).

9. National Research Council of the National Academies [A Framework for K-12 Science Education: Practices, Crosscutting Concepts and Core Ideas] National Academies Press, Washington, DC, foreward, 8, 10, 41-42 (2012).

10. Popejoy, A., “American Women in Science,” New Voices for Research, 09 March 2011. <http://newvoicesforresearch.blogspot.com/2011/03/american-women-in-science.html> (16 April 2012).

11. “U.S. News 2012 Best Engineering Schools Rankings and Reviews,” U.S. News, 2012. <http://grad-schools.usnews.rankingsandreviews.com/best-graduate-schools/top-engineering-schools/eng-rankings> (16 April 2012).

12. “Bergen County Academies: Academy for The Advancement of Science & Technology Class of 2012 College Profile, ” Bergen County Academies, March 2012. <http://bcts.bergen.org/images/stories/BCTS/pdf/AAST_profile%202012%20updated.pdf> (16 April 2012).

Acknowledgements• Alyssa Calabro, C.E.M.T• Dr. Howard Lerner, Superintendent of the Bergen County Technical School

District • Edmund Hayward, Director of Technology for the Bergen County Technical

School District• Russell Davis, Principal of the Bergen County Academies• The Administration of the Bergen County Technical School District• The Administration of the Bergen County Academies• The Bergen County Technical School District Board of Education• The Bergen County Board of Chosen Freeholders• The research faculty members: Dr. Robert Pergolizzi, Donna Leonardi, and

Dr. Deok-Yang Kim • Our collaborating research partners

• For more information, visit our website: www.bergen.org/nsil