Enhancing senior high school student engagement and academic performance using an inclusive and scalable inquiry-based program

The multi-disciplinary nature of science, technology, engineering, and math (STEM) careers often renders difficulty for high school students navigating from classroom knowledge to post-secondary pursuits. Discrepancies between the knowledge-based high school learning approach and the experiential approach of future studies leaves some students disillusioned by STEM. We present Discovery, a term-long inquiry-focused learning model delivered by STEM graduate students in collaboration with high school teachers, in the context of biomedical engineering. Entire classes of high school STEM students representing diverse cultural and socioeconomic backgrounds engaged in iterative, problem-based learning designed to emphasize critical thinking concomitantly within the secondary school and university environments. Assessment of grades and survey data suggested positive impact of this learning model on students’ STEM interests and engagement, notably in under-performing cohorts, as well as repeating cohorts that engage in the program on more than one occasion. Discovery presents a scalable platform that stimulates persistence in STEM learning, providing valuable learning opportunities and capturing cohorts of students that might otherwise be under-engaged in STEM.


Introduction
The goal of IBBME is to provide students with a real challenge in biomedical engineering, for which an introductory skill lab provides them with the technical ability to address the challenge in some way. In addition to the subject-specific technical skills that students will gain, the curriculum is also designed to give students highly transferable general scientific and engineering skills including: For each subject (i.e. biology, chemistry, and physics), we have included: • An outline of the research project for the purposes of logistical and curricular planning, and indicating the nature of the challenge, research question, expected methodology, and learning outcomes for students • A skill lab protocol in the style of a standard 3-hour undergraduate laboratory, adjusted for technical and theoretical level and designed to introduce experimental and analytical techniques necessary to complete the conceived project • A request for proposal (RFP) presented to the students outlining the motivation underlying the biomedical engineering challenge that the students are expected to contribute to, and to which the students must respond with a formal proposal S2 Appendix: Sample teaching materials for one term of IBBME Discovery 4 At the beginning of the skill lab, students were required by the facility to undergo a brief safety training that covered considerations of the facility in general, as well as safety procedures specific to the techniques and equipment in use and standard emergency procedures. Material safety data sheets (MSDS) for chemicals used were made available to all participants before the day of the skill lab. All graduate instructors participated in all department-mandated safety training modules.
After safety training, students typically took part in a 30 to 60 minute interactive lecture pertaining to the grand challenge of the semester, as well as introducing the theory and technical knowledge required to complete the skill lab protocol for the rest of the day. Students had been provided the skill lab protocol before attending, and they were given the rest of the day to complete the protocol as written, with instructor assistance as needed. At the end of the day, the RFP outlining the challenge was distributed to the students, and time was taken to discuss with students the nature of the project and potential avenues of inquiry.
Here we have provided sample documents for a past semester of Discovery, including outlines, skill labs, and RFPs for all three subjects. We have also included the template students use to complete the proposed research plan. The central theme of the semester was "Cardiovascular Disease: Engineering diagnostics and treatments". This served to constrain project directions, rationalize the skills and techniques covered, and motivate students.

Biology
Biology project outline Motivation: Currently in Canada there are over 1.6 million people with cardiovascular disease, and cardiac health is one of the leading causes of death in the developed world. These patients suffer from a significant reduction in their quality of life as reduction in heart function makes everyday life difficult.
Although healthcare continues to advance treatment, there is a need for continued focus on preventing, identifying and treating this prominent health problem.
Cardiotoxicity effects are the leading cause of pharmaceutical drug withdrawal from markets; and delays in regulatory approval. Thus, cardiac safety assessment has become an important regulatory step in drug development. Inappropriate drug formulations or excessive drug exposure can lead to cardiac toxic effects that include arrhythmia, cardiac electrical instability, and sudden cardiac death. Some of these toxicity effects can be regulated in a time-dependent and dose-dependent manner. We want to investigate the cardiotoxicity effects of an unknown chemical compound, Substance X; however, we will require the expertise of cell biologists to conduct experiments comparing Substance X to known cardiac toxic drugs.
Research question: How can we characterize the toxic effects of chemical compounds on cardiac cells?
Rationale: Knowledge of cardiac cell characteristics (beating frequency, morphology, calcium oscillations, etc.) allows us to distinguish between cardiac cells that are functioning normally and cardiac cells that are beginning to fail. Once there is an understanding of the differences between healthy and dysfunctional cardiac cells, we can start to evaluate the potential cardiac toxicity effects of chemical compounds on cardiac cells.

Skill-based activity:
A full day of introductory programming will include: • Laboratory Biosafety (lecture, demonstration, quiz) • Introduction to Pipetting & Serial Dilutions (lecture, pipetting practice) • Cell Morphology and Microscopy (lecture, microscopy demo, cell imaging) • Cell Staining -live/dead, DAPI (lecture, cell imaging) Required equipment: Students will be given plates of cardiac cells, access to fluorescence microscopes, and a chemical inventory to characterize the cardiac tissue and explore how cell characteristics are impacted by various chemical compounds.
Expected outcomes: By the end of the module, students will be able to: • Become familiar with basic laboratory practices and techniques 5. Repeat this serial dilution with tubes # 3-8.

What is the concentration of dye within each tube? What happens to the colour of each solution?
Protocol: Cell culture and microscopy practice For today's lab, you will be working with HeLa cells. You aren't expected to know the answers to the questions in the protocol ahead of time. Feel free to discuss with your instructors, and make sure to ask any questions you have. There are no stupid questions in a biology lab! What do you know about these cells? Where do they come from?

Cell Culture
1. Obtain two dishes of cells from your instructor. Label them both with your group number or names, and mark one as "control" and one as "room temperature".
2. Replace the cell culture media in both dishes as directed by your instructor, using sterile technique. Return the "control" plate to the incubator and leave the "room temperature" plate in the sterile flow hood before you go to lunch.

Based on what you know about cell culture, what do you think will happen to these cells? How could you test this hypothesis?
3. When you return from lunch, obtain the stock solution tubes of ethidium homodimer (EthD) and calcein-AM from your instructor. Make sure to leave the aluminum foil on the tubes.

Why is it important to shield these solutions from light?
4. Using sterile technique, add 5 µL of EthD and 2.5 µL of calcein-AM to 5 mL of phosphatebuffered saline (PBS) in a new 15 mL Falcon tube. This is your dye solution.
5. Remove the cell culture medium from both plates and add 2.5 mL of your dye solution to both plates. Allow them to incubate for 30-45 min.

Fluorescent Microscopy
1. With the instructor's help, look at your cells using the microscope. Image your plates using brightfield, phase contrast, and red and green fluorescent channels. Record representative images (at least 3 replicates per plate) in all modes using both 10X and 40X objectives. Note that you need to capture all channels in the same place before moving your sample. 4. Cell counting: From your 10X images, use a particle counting tool (Plugins > Analyze > Cell Counter) to count how many cells are fluorescing red, and how many are fluorescing green.
Calculate the percentage of dead cells for each sample. Cell counting: Use the plugin "Cell counter" to count cells in your images. Open the plugin: Plugins>Analyze>Cell counter and then click on your image, making it active. Click initialize in the plugin window, select a counter type and make sure you have one of the selection tools active (e.g. click on the "freehand selection" tool in the ImageJ toolbar). Click on all cells of interest.
If you make a mistake, you can press "delete" in the plugin and the last click of that type will be removed.
If you check the box "delete mode", you can click in the image to delete the closest marker of that type.
Obsolete: In newer installations of ImageJ, the Cell Counter plugin has been removed and is replaced by the native Multipoint Tool which is even easier to use. The plugin should be available on all lab computers but if you install ImageJ at home, this might not be the case. To run the multipoint tool, simply launch it from the toolbar, select a colour and cell type number and click away. (Alternately, try to open the plugin as normal and ImageJ will automatically launch Multipoint tool for you.)

Tracing cells:
Wand tool: Using the Wand Tool, you can click on a pixel and all connected pixels of similar intensity will be selected. This is good if there is high contrast between cells (e.g. a darker area between). If you double click the wand tool, you can try different methods to determine "neighbouring" pixels and you can set tolerance for variations. A tolerance of 0 means pixels need to have the same intensity to be selected.
Increasing the tolerance gives more leeway and pixels of similar intensity will be selected. This can be done live by selecting a cell and moving the slider. These modifications can be combined with the wand tool: select a cell using the wand tool and add or subtract areas with the freehand tool. .

Biology request for proposal
Currently in Canada there are over 1.6 million people with cardiovascular disease, and cardiac health is one of the leading causes of death in the developed world. These patients suffer from a significant reduction in their quality of life as reduction in heart function can make everyday life difficult. Although healthcare continues to advance treatment, there is a need for continued focus on preventing, identifying and treating this prominent health problem. We are looking for the next generation of great minds to help us improve the lives of these people around the world! Cardiotoxicity effects are the leading cause of pharmaceutical drug withdrawal from markets and delays in regulatory approval. Thus, cardiac safety assessment has become an important regulatory step in drug development. Inappropriate drug formulations or excessive drug exposure can lead to toxic effects including arrhythmia, cardiac electrical instability, and sudden cardiac death. Some of these toxicity effects can be regulated in a time-dependent and dose-dependent manner. We want to investigate the cardiotoxic effects of a newly developed chemical compound, Substance X. However, we will require the expertise of cell biologists to conduct experiments comparing Substance X to known chemicals and/or drugs.
We will be collecting proposals from multiple teams and will consider all options at a collective symposium. Please submit your project proposal using the RFP template provided. We have attached a list of the materials available in our facilities, so please take a look at these as you build your experimental plan. These will be reviewed by IBBME representatives who will meet with you to finalize the experimental details. You will then utilize our facilities to carry out your project and deliver a final presentation in poster format (template provided). We are looking forward to seeing the innovative solutions you will develop! S2 Appendix: Sample teaching materials for one term of IBBME Discovery Although healthcare continues to advance treatment, there is a need for continued focus on preventing, identifying and treating this prominent health problem.
Metoprolol is a beta-blocker that is frequently used to treat high blood pressure and tachycardia (chronically elevated heart rate). It can be taken by pill or intravenously but is a "dirty drug" in that it has numerous side effects, including depression, insomnia, fatigue, dangerously low heart rate, or respiratory depression, among others. Part of this toxicity can be avoided by tightly regulating the concentration of the drug within the body. We are working on bringing a new metaprolol treatment to market in either pill or implant form but need chemical engineering expertise to optimize the release of the drug within safe limits.
Research question: Can a hydrogel's properties be engineered to release a drug within safe limits over time?
Rationale: Knowledge of cardiac cell characteristics (beating frequency, morphology, calcium oscillations, etc.) allows us to distinguish between cardiac cells that are functioning normally and cardiac cells that are beginning to fail. Once there is an understanding of the differences between healthy and dysfunctional cardiac cells, we can start to evaluate the potential cardiac toxicity effects of chemical compounds on cardiac cells.
Skill-based activity: Students will compare prepared gelatin to alginate beads in their release kinetics of food colouring using a spectrophotometer and standard curve. Knowledge of the Beer-Lambert law and S2 Appendix: Sample teaching materials for one term of IBBME Discovery 19 chemical equilibria will be necessary. Students will gain experience with all relevant techniques and will learn to use Excel graphing to draw quantitative conclusions. Students will use FCF Brilliant Blue #1 (blue food colouring) for all activities. First, they will be required to create a serial dilution series of stock dye to find a dilution suitable for reading in a spectrophotometer. By using a known extinction coefficient, they will be able to calculate the stock concentration. Next, they will need to create a 5-point linear standard curve within the optimal range of the spectrophotometer based on their understanding of the relationship between absorbance and concentration. Finally, they will use this curve to measure the release of Blue #1 from premade alginate beads into media over time.
Project: Students will create hydrogels from alginate and/or gelatin and measure their release of blue dye over time. They will be able to modulate gel composition and density, form (size/shape), and initial amount of dye loaded. Using a standard curve and timepoint measurements, they will generate release profiles that will be loaded into a premade mathematical model of metoprolol absorption and clearance from the human body. This will allow them to assess the efficacy and safety of their hydrogel drug delivery system. • Pipettes

• Spectrophotometer
Expected outcomes: At the end of this module, students will be able to: • Apply concepts and use common equations in equilibrium and the Beer-Lambert Law.
• Understand the theory and limitations of a spectrophotometer.
• Understand hydrogel structure and function.
• Design a proposal to tailor the rate of drug release within a specified range, based on modifying common material properties of a hydrogel (size, shape, polymer content, starting concentration).

Relevant literature topics:
Custom protocols and presentations will be provided. Readings on equilibrium, conservation of matter, etc. in their own textbooks will be helpful for review. S2 Appendix: Sample teaching materials for one term of IBBME Discovery 21

Chemistry skill lab
To be completed during lecture: Metoprolol is a beta-blocker. What is the role of a beta-blocker in cardiac medicine?
What are three of the side-effects of metoprolol?
In general, how can we avoid side-effects while gaining desirable effects of a drug?

What properties of a hydrogel can be changed to tune drug release?
Today's activity: Today we will be modeling drug concentrations and controlled release using a blue dye (FCF Brilliant Blue #1, or blue food colouring). Blue #1 shows peak absorbance at 628 nm, has a molar mass of 792.84 g/mol, and an extinction coefficient of 130 000 L/mol/cm.

Sketch a plot of absorbance vs. wavelength for Blue #1:
Why should we measure absorbance at the peak wavelength?
1. Place 10x 1.6 mL epitubes in a rack. Add 2 drops of blue food colouring to one. This is your working dye stock solution.

What is the dilution of this solution in relation to the original dye stock?
Why should we use ultrapure water for spectrophotometry? 11. Add 200 µL from each tube to a well in your microplate, so that B1, B2, and B3 hold the triplicate samples made in step 8; C1, C2, and C3 hold the triplicates made in (9), etc.

Chemistry request for proposal
The Institute of Biomaterials and Biomedical Engineering (University of Toronto) is working to improve the lives of patients around the world. Currently in Canada there are over 1.6 million people with cardiovascular disease, and cardiac health is one of the leading causes of death in the developed world.
These patients suffer from a significant reduction in their quality of life as reduction in heart function makes everyday life difficult. Although healthcare continues to advance treatment, there is a need for continued focus on preventing, identifying and treating this prominent health problem. We are looking for the next generation of great minds to help us improve the lives of these people around the world! Metoprolol is a beta-blocker that is frequently used to treat high blood pressure and tachycardia (chronically elevated heart rate). It can be taken by pill or intravenously but is a "dirty drug" in that it has numerous side effects, including depression, insomnia, fatigue, dangerously low heart rate, or respiratory depression. Part of this toxicity can be avoided by tightly regulating the concentration of the drug within the body. We are working on bringing a new metaprolol treatment to market in either pill or implant form, but need chemical engineering expertise to optimize the release of the drug within safe limits. We want you to design a metoprolol-releasing hydrogel and quantify its rate of release to fall within safe limits.
We will be collecting proposals from multiple teams, and will consider all options at a collective symposium. Please submit your project proposal using the RFP template provided. We have attached a list of the materials available in our facilities, so please take a look at these as you build your experimental plan. These will be reviewed by IBBME representatives who will meet with you to finalize the experimental details. You will then utilize our facilities to carry out your project and deliver a final presentation in poster format (template provided). We are looking forward to seeing the innovative solutions you will develop!

Notes:
Be aware of experimental constraints You will have access to calcium and alginate solutions, food dye, materials to cast gels, microplates and spectrophotometers, and potentially other chemicals or materials available upon request.

Do your research
• How can we modify hydrogels to change delivery of a drug?
• What changes to current treatments would patients benefit from?

Engineering is an iterative and creative process
Some ideas you have might not be directly in line with the precise problem posed. There are plenty of issues in healthcare today, and given appropriate resources, you can adapt the project to your interest!

If your proposal is approved, be prepared to make it happen!
• Have a step-by-step protocol ready • Be prepared to troubleshoot difficulties as they arrive Although healthcare continues to advance treatment, there is a need for continued focus on preventing, identifying and treating this prominent health problem.
Improved resources for patients with cardiovascular disease to monitor their conditions would allow them to track parameters of interest, and schedule appointments or alert emergency services as needed if key metrics of cardiovascular performance change. A portable infrared plethysmograph capable of tracking pulsatile blood flow can be easily built using simple Arduino-compatible components.
Research question: Can we build a cost-effective infrared pulse sensor device to monitor cardiac function?
Rationale: Biomedical instrumentation is necessary to monitor and diagnose medical conditions.
Students will learn the scientific basis of biological signals and the technical principles involved in detecting and measuring them. Close monitoring of a condition is also necessary for optimized intervention.
This project would also complement the "Waves and Sound" and "Electricity and Magnetism" units from the grade 11 and 12 physics curricula: Skill-based activity: Several small tasks will teach the students some of the following concepts: • Fundamentals of circuits: basic electronic components, units and concepts associated with electricity, and voltage/current laws.
• Signal Processing: signal filtering and amplification • Coding: Writing code to calculate and display heart rate The skill-based learning activity will involve a guided-session where students are introduced to the Arduino, a popular microcontroller, which is effectively a very simple computer. Students will follow steps to setup the Arduino, learn about its various components and features and how to upload code to the Arduino. At the end of the skill laboratory, students will have created a simple circuit interfaced with the Arduino that allows them to control the blink frequency of an LED by adjusting a potentiometer (a variable resistor device). This will teach skills including basic circuit design, collecting signals into the Arduino (a process known as analog-to-digital conversion) and how to send signals from the Arduino to an LED.
Project: Students will create a prototype of an IR pulse sensor/plethysmography device for measuring pulsatile blood flow in an extremity. This device and the process of creating this device will expose students to biomedical device design and development. By combining the scientific principles of cardiac physiology and engineering students will see first hand how scientific research can be translated into an engineering device that can be used to improve patients' quality of life.

• 3D Printers
Expected outcomes: At the end of this module, students will be able to: • Understand basic electronic components and be able to plan and assemble rudimentary electronic circuits • Obtain, modify, and run basic Arduino code • Understand basic data acquisition and filtering processes • Understand principles of 3-dimensional CAD and additive manufacturing Relevant literature topics: Custom protocols and presentations will be provided. Readings on equilibrium, conservation of matter, etc. in their own textbooks will be helpful for review.

Physics skill lab
Purpose This lab will introduce several important microcontroller concepts and features. First, it introduces the concepts of inputs and outputs. Inputs can allow the Arduino to connect to sensors, which detect variables from its environment (such as temperature) or allow a user to interact with it through buttons or a potentiometer. On the other hand, outputs allow the microcontroller to control other components or devices in order to display information back to the user or run a system. In this lab, we will see how programming the microcontroller allows us to control the relationship between those inputs and outputs.

Materials
• PC + Arduino IDE • Arduino microcontroller + USB cable 1. Connect the positive rail on the breadboard to +5V on the Arduino and the ground rail to any GND pin on the Arduino.
2. On the breadboard: Following the diagram in Figure 1, connect the digital output pin D13, the 100Ω resistor, the LED, and the ground rail in series. Note that the LED has polarity a .
S2 Appendix: Sample teaching materials for one term of IBBME Discovery  These patients suffer from a significant reduction in their quality of life as reduction in heart function makes everyday life difficult. Although healthcare continues to advance treatment, there is a need for continued focus on preventing, identifying and treating this prominent health problem. We are looking for the next generation of great minds to help us improve the lives of these people around the world! Early detection of abnormal heart rhythms is important in order to provide timely and appropriate treatment for cardiac conditions. There are two main techniques that can be used to monitor a person's heart rhythm. These include measuring the electrical activity of the heart (an electrocardiogram) or detecting changes in the amount of oxygenated blood within a tissue region (e.g. a fingertip) based on the amount of infrared (IR) light that is reflected or transmitted (known as IR plethysmography). This works because the oxygenated blood contains higher levels of oxy-hemoglobin (hemoglobin is a compound in red blood cells which transports oxygen around the body), which reflects more infrared light than deoxygenated blood. We are looking for a cost-effective device to measure a patient's heart rate.
The design should be able to be easily prototyped and demonstrate the conceptual design of the device. The device should be comfortable, easy-to-use, and portable. It should be able to indicate the user's heart rate and alert them if there are measured abnormalities.
We will be collecting proposals from multiple teams and will consider all options at a collective symposium. Please submit your project proposal using the RFP template provided. We have attached a list of the materials available in our facilities, so please take a look at these as you build your experimental plan. These will be reviewed by IBBME representatives who will meet with you to finalize the experimental details. You will then utilize our facilities to carry out your project and deliver a final

Do your research
• Why is it important to monitor heart rate?
• What are some techniques that can be used to measure heart-rate?
• Compare the two most common techniques. What are their advantages and disadvantages?
• What is IR plethysmography (photoplethysmography)? How does it work?
• What are normal ranges for heart-rate?
• How can the device give feedback to the user?

If your proposal is approved, be prepared to make it happen!
Have detailed protocols ready and be aware of safety considerations around fabrication and electronics.

Research Proposal Template
The following is a condensed version of the proposal template students complete in response to their subject specific RFP each term. Blue text is descriptors that guides student writing into specific categories, which is removed prior to submission.

TEMPLATE INTRODUCTION
Use this template to respond to the "Request for Proposal" from IBBME Discovery, thereby developing a Experimental groups that will be considered.

RESPONSIBILITIES IN PROJECT MANAGEMENT
This section outlines when you will achieve each part of your proposed project and who is responsible for

S3 Appendix: Mark breakdown for student assessment in Discovery programming
Enhancing senior high school student engagement and academic performance using an inclusive and scalable inquiry-based program The following

CLIENT MEETING
Student presentation skills in virtual client meeting that pitches proposed workflow in response to subject specific request for proposals 5 3.4%

FINAL PROPOSAL
Group submission of planned workflow for the skill lab visits. Assessment of workplan completeness, appropriate research question, hypothesis, and expected outcomes. Description of groupwork breakdown clearly described 35 24.1%

PROGRESS REPORT
Reflection on first skill lab visit outcomes, summary of progress to date Updated plan of next steps to ensure research outcomes are complete at the end of next skill lab visit

S4 Appendix: Entrance and exit student surveys for program assessment of IBBME Discovery
Enhancing senior high school student engagement and academic performance using an inclusive and scalable inquiry-based program