Questioning the Research Question: A Collapsing House of Cards

Hey readers!

After some extensive meetings with Mrs. Haag last week (thank you so much, Mrs. Haag), I figured out how to organize my literature review in a logical and cohesive order compared to my previous blogpost. I came into this class with a set way of thinking about my research from my previous work, which was governing how I wrote my outline. I was having difficulty considering how someone new to the research needed the information presented to understand it.

RIP Bird Gang

Much like the Cardinals this past week, I felt like I had so many chances to win -- to make a great outline -- but I fell short. I threw an interception in the red zone (I'm looking directly at you, Carson Palmer). I couldn't pick up a forced fumble and ended up kicking it out of bounds (Ehem, Tyrann Mathieu). At least I didn't have any fans to disappoint (Wait, but are you, my blog readers, the fans in this complex and extraneous metaphor? In that case, I'm sorry!). Now, the focus moves on to redemption -- the LA Rams and an official literature review outline due 10/7.

In the meantime, I am forming my preliminary research question based on the information I have compiled and the gaps in the research I have found. Here's what I envision for my research question: "To what extent is HemaDrop™ a viable method for determining blood composition by making a uniform, solid, and analyzable film from microliters of blood?"

Here why I decided on this question:

1. Scope: The scope of my research has been quite narrow from the beginning, as I am working  on a specific technology (HemaDrop™). Additionally, this research doesn't start out with the aim to create the perfect formulation of HemaDrop. Instead, the goal is to assess the efficacy of the technology in its current form and provide potential improvements based on the conversation between the literature and the results of the feasibility experiments. In this way, the final product of my research will not be judged exactly on if I create a successful product for microliter blood analysis, as much more will be required than the scope of an AP Research project. 
2. Key Terms:
• HemaDrop™: method for solid-based microliter blood analysis 
• Viable: Provide necessary quantitative information about blood composition including elemental and molecular composition, as a flexible method for creating samples for different techniques. 
• Uniform: different spots on the sample should not yield different compositions. 
• Solid: Only solid films are analyzable in vacuum, which is what most techniques require 
3. Variables: For my experiments, I am testing the viability of the analysis based on the quantitative results. This is the response variable. In my research, the explanatory variable are the properties of the substrate (composition, coating, etc) and the analysis technique. Based on the substrate properties, once the composition of the blood or saline solution is determined, the composition values of they are known could be used to compute the accuracy of the measurements, along with the error. 
4. Research-ability: I had some researchability concerns about the amount of analysis techniques I would have to look into, since I envision my methods creating a bunch of samples prepared in different ways (e.g., type of coating, substrate, etc.). There would be far too many combinations to test. For this reason, after talking to Dr. Herbots (my mentor), I will test 3, or a maximum of 4, analysis techniques (the best ones I determine based on the literature review). I have several sources which describe the advantages and limitations of the analysis techniques. By limiting the number of analysis techniques I will examine, I can make sure that the experiments I conduct are not too many, but also capture the full complexity of the research to come to an insightful conclusion. I see myself answering the question by comparing the results (accuracy, precision, etc) of using analysis techniques on various samples with canine blood, saline, and human blood. The comparison of these techniques will demonstrate how viable HemaDrop substrates are for making a uniform film of blood that can be analyzed by a variety of techniques.
5. Gap: There has been no successful implementation of microliter blood analysis thus far. Theranos has attempted liquid microliter blood analysis, but failed. The limitations of milliliter blood analysis create a clear need for microliter blood analysis, so a clear gap exists in the biomedical industry and in the care for patients. This research aims to fill this void and successfully perform microliter blood analysis. By applying concepts like superhydrophilicity and vacuum-based analysis techniques, HemaDrop aims to create a uniform analyzable film of blood from drops.
6. Significance: The signficance of this research is the creation of microliter blood analysis. The reduced volumes of blood is reduced discomfort in patients who undergo blood testing. Additionally, the problem of hospital-acquired anemia will also be mitigated in critically ill patients. By using less blood, doctors also can test patients more often to assess their diagnosis and monitor their conditions. Finally, the cost of microliter blood analysis will be less than milliliter blood analysis. 

Let's get back to the NFC Championship Game and back to rolling with research!

Cheers,

YP

(863)

The Outline of a Lit Review

Hey readers!

Now that I have compiled about 15 sources about HemaDrop, it's time to start actually outlining my literature review -- it's all business. In case you forgot, I have been using the following subtopics to classify my sources: (1) Blood diagnostics, (2) Superhydrophilic films, (3) Blood Properties, (4) Substrates, (5) Analysis Techniques, and (6) Key Illnesses/Conditions.

Here's what I envision for my literature review:

Subtopic 1: Blood Diagnostics

I have to discuss blood diagnostics in general to introduce the general field of research I am investigating and describe its importance before talking about mechanisms behind blood analysis. Additionally, by explaining the current technology used for blood testing, I can highlight its evolution, needs, and limitations -- basically the gap.

• Premise 1: Blood testing is crucial for diagnosing illnesses, monitoring the conditions of patients, and providing healthcare to patients.
• Barthels, et al: Blood testing is the cornerstone of medicine, as it has allowed for diagnosing of many conditions based on markers in the blood.
• UK National Health Service Database of Illnesses and Blood Markers: E.g., conditions like diabetes, anemia, cancers, heart attacks, etc. all alter blood composition in a specific way, including increased glucose content, lack of Fe, presence of specific biomarkers, elevated troponin concentrations.
• Premise 2: Current blood tests require 7 mL of blood on average.
• Barthels, et al & Hamerling
• Examples include high-performance liquid chromatography (HPLC), spectroscopy of liquid vials of blood, testing conductivity of blood for hematocrit levels, etc.
• Premise 3: The current volume of blood required for testing (7 mL) causes patients discomfort, complications including hospital-acquired anemia (HAA), and can limit the number of tests possible for ill patients (thereby preventing diagnoses too).
• Bom, et al: Better testing procedures and advancements in diagnostics is necessary to stop HAA and unnecessary costs on the healthcare system.
• Thakkar, et al: Current blood diagnostic testing causes HAA, incurs unnecessary cost on the healthcare system, and reduces the quality of patient care. 
• Premise 1 + Premise 2 + Premise 3 --> Conclusion 1: A need exists for blood diagnostic technology that uses smaller volumes of blood to improve patient care, prevent HAA in critically ill patients, and reduce costs.
• Premise 4: Due to the importance of blood analysis, need for standardization, and ease of use needed for doctors and nurses, blood analysis techniques must: "use whole blood and a small sample volume, provide results in less than 1-15 min, maintain high accuracy (at least at current levels), have simple operation and inbuilt quality control, require minimal maintenance,  and use an IT interface."
• Barthels, et al: Point-of-care testing with glucose monitors and similar devices that can provide real-time and accurate monitoring of conditions are part of the next generation of medical devices.
• Premise 5: Previous attempts at blood analysis with smaller volumes (e.g., Theranos) have been unsuccessful.
• Kidd, et al: Theranos had errors of over 10%, the standard for biomedical testing
• Potentially could find more examples?
• Conclusion 1 + Premise 4 + Premise 5 --> Conclusion 2: If implemented, small volume blood analysis (microliter) must meet the aforementioned criteria.
• Premise 6: Both solid and liquid types of blood analysis exist, with liquid requiring little processing and potential lack of uniformity, but with solid allowing for vacuum-based analysis techniques and use of smaller volumes.
• Pershad, et al
• UK National Health Service Database of Illnesses and Blood Markers
• Conclusion 2 + Premise 6 --> Conclusion 3: Potentially a solid-based blood analysis technique, if it overcomes issues with processing time and uniformity, could use small volumes of blood for analysis with vacuum-based techniques.

Subtopic 2: Superhydrophilic Films

The mechanism behind the uniform, thin, solid film that HemaDrop creates is superhydrophilic films (or hyper-hydrophilic if you wish). The explanation of this technology follows naturally after the potential of solid-based blood diagnostics is introduced, since this property is what makes HemaDrop create a uniform film. 

• Premise 7: Superhydrophilic films are created by applying superhydrophilic coatings on surfaces to make them attract water.
• Wen, et al: Superhydrophilicity characterized by high contact angle and high CA hysteresis change by sliding angle -- SHFs can be created by high surface energy coatings and reducing surface roughness (relates to substrates) 
• Hosokawa, et al: "Extremely high receptivity to water adhesion" 

• Premise 8: Superhydrophilic films, because of the importance of polarity have a wide range of biomedical applications
• Hosokawa, et al
• Drelich, et al:
• "For example, surfaces of hydrophilic materials were roughened in the past to improve adhesion in composites, biocompatibility in implant devices, or simply to enhance spreading of liquid, even so these activities were not linked yet to superhydrophilicity" 
• Premise 9: The wettability/hydroaffinity of the surface on which a blood drop affects its uniformity and phase separation.
• Need more support for this -- maybe look at chromatography methods which use a polar mobile phase. (Note: I didn't think of this claim earlier, so good thing I'm outlining!)
• Subpremise 3.1: Since some elements of blood (e.g., the lipid cholesterol) are non-polar, phase separation will occur on hydrophilic samples between non-polar and polar elements.
• Premise 7 + Premise 8 + Premise 9 + Conclusion 3 --> Conclusion 4: The hydroaffinity of a surface can be manipulated to make it superhydrophilic before a small drop of blood is applied. The surface's properties (e.g., surface energy from hydrophilicity) will affect how the blood dries, potentially making the film uniform and analyzable (combatting a major problem with solid analysis of blood)
• Potential Gap -- Determining the optimal degree of wettability to minimize phase separation and heterogeneity, maximize analyzability

Subtopic 3: Blood Properties

Now that we have looked at the coating, we have to investigate the properties of what is actually drying on the surface. Specifically, properties of blood which prevent uniform drying and solid state analysis should be investigated, as they could affect accuracy of measurements or even prevent them.

• Premise 10: Blood contains cells of varying types (e.g., RBCs, WBCs, etc.) that all separate by hydroaffinity and density.
• Palta, et al.
• Can stay unrefrigerated for 24 hrs without separating.
• Premise 11: Blood coagulates in vitro very differently than in vivo, as it uses a cascade of proteins including thrombin, fibrinogen, and protein S, which are activated by Ca ions.
• Acharya, et al.
• Premise 12: Superhydrophilic films limit the aforementioned effects on the blood while preparing a HemaDrop sample.
• Hosokawa, et al & Pershad, et al.
• Premise 10 + Premise 11 + Premise 12 --> Conclusion 5: SHFs can control the natural tendencies of blood to coagulate and separate on surfaces.

Subtopic 4: Substrates

Substrates, although they end up being coated, are extremely important, as many analysis techniques (including Rutherford Backscattering Spectrometry) are affected by conductivity of the sample. Additionally, for commercialization, cost and ease of use are necessary considerations when preparing a sample for analysis. Thus, the, cost, conductivity, and other properties of the sample must be compared to determine the optimal sample type.

• Premise 13 (branching off Premise 7): The hydroaffinity of a surface is affected by the surface topography (roughness). not just the coating, so the substrate affects the degree of superhydrophilicity we are able to achieve.
• Wen, et al.
• Hosokawa, et al.
• Premise 14: Tests have been conducted on TiN/Si(100) and Si(100) surfaces using canine blood with and without the superhydrophilic coatings.
• Pershad, et al: Results published demonstrated a clear difference and affect of the SHF, but the substrates were expensive to use due to their lack of major topographical features (they were smooth)
• Premise 13 + Premise 14 --> Conclusion 6: A more cost-effective alternative, including microscope slides (e.g., borosilicate), should be investigated and the resulting hydroaffinity should be compared after treatment with SHF.
• Potential Gap -- The effectiveness of these more rough surfaces should be tested as an alternative to smoother wafers as tested in Pershad, et al.

Subtopic 5: Analysis Technniques

After looking at the sample preparation mechanisms, it is natural to look into how the samples should be analyzed. A comparison of the limitations, strengths, and properties of various microscopies and spectroscopies will be undertaken to determine which ones to investigate with HemaDrop during experimentation.

• Premise 15: Solid blood samples that are prepared uniformly on substrates can be analyzed in a vacuum, allowing for many applicable microscopies and spectroscopies.
• Mukhopadhay, et al: Describes the conditions necessary for analyzing samples in vacuum
• Premise 16: Atomic Force Microscopy, Scanning/Tunneling Microscopy, IR Fourier Transform Spectroscopy, Ruthterford Backscattering Spectrometry, Particle-Induced X-Ray Emission, X-Ray Photoelectron Spectroscopy, etc. are possible analysis techniques.
• Here's where I realize I need more sources to compare them -- I was having trouble finding good sources that have comparable information about all of them.
• Gap -- which techniques to consider? -- no conclusion yet.

Subtopic 6: Key Illnesses & Conditions

Finally, I want to investigate the specific markers in the blood of several illnesses and potentially simulate the disease in samples to determine whether the difference would be detected. To do this, I found a large database from the UK National Health Service. There are over 1000 illnesses, so to finish off I need to select which illnesses I want to specifically. I don't really have premises here, as I have just looked at diabetes, anemia, various types of cancer, and the monitoring of specific antibiotic concentrations as specific paths to test. I need to find studies who investigated diseases in a similar way that I am, perhaps after they found a new method of analysis.

My research question will then be something along these lines: "Can HemaDrop successfully create a uniform, solid, thin film and accurately analyze blood composition from microliters of blood?” Due to issues with IP, I can look into testing HemaDrop's viability rather than developing the product itself.

Well, I started writing and got very into it. So, there it is. Also, I apologize for no humor. I got too focused on content. Hopefully, I can start attacking the question parts I have outlined. I think if I can fill in those gaps, I'll have a solid understanding of what's going on in my field of study.

Cheers,

YP

(1, 598 -- RIP word limit)

Hospital-Acquired Anemia: A Text Convo

Hey readers!

This week, in class, we looked at John Oliver's most recent episode about charter schools, broke down his argument, and are creating our rebuttals against his points. Apart from the experience being very entertaining, I also enjoyed it because I realized that when I listened to John Oliver, I often simply assumed he was always right because I love watching his show. I guess AP Seminar skills have to applied at all times... My guard will now be up at all times!

Anyways, this week in my research project, I reviewed a lot of sources about current errors in laboratory diagnostics and technology. I also talked to one of my relatives who is a pathologist in the Phoenix area, and she said that I could come tour the laboratory where she works sometime in the next 2 weeks, which is very exciting for my literature review. It has been really hard to find exactly what instruments/techniques hospitals and laboratories are currently using for tests, since they vary so much. However, by looking at what currently is being used, I can establish a criteria for precision, speed, and form for analyzing HemaDrop samples. Since HemaDrop must be at least as good as what is currently available to be viable.

Two sources that I investigated were very very in conversation with each other. They concerned the issue of hospital-acquired anemia (HAA). The relevance of HAA to microliter blood analysis is that HAA often occurs in very sick patients at the hospital, since they require lots of blood testing to diagnose and monitor their conditions. However, these two sources, one a response to another, provide contradicting explanations of the causes of HAA. I somehow got access to their personal, private, and heated text conversation! Here it is! The links to the articles are here (Hospital-acquired Anemia: The Contribution of Diagnostic Blood Loss by Dr. Van der Bom, et al.) and here (Impact of an Educational Intervention on the Frequency of Daily Blood Test Orders for Hospitalized Patients by Dr. Rajiv Thakkar, et al.).

So it seems like these two contradicting sources agreed on the gap in the research -- HemaDrop! I think this conversation is really interesting because it describes the goals of all biomedical research: getting more information from less invasive procedures.

Hope you enjoyed the exclusive convo! I am very excited for this week as I will be hearing back from the pathologist, and I found a great source on analysis techniques.

Cheers,

YP

(673 -- including the text messages, which I wrote as well)

Lit Lit Review: Mini-dabbing or Mini-blood Analysis?

Hey readers!

Happy Labor Day! And we all know Labor Day means two things -- football is back, and it's time to review some literature...

This baby's love of football equals my love of literature reviews?

This week, we learned the essentials of a literature review, I worked on compiling and reading some sources, and the legend of the mini-dab was created. Since both microliter blood analysis and mini-dabbing are compact, smaller, and better versions of original blood analysis and dabbing respectively, mini-dabbing is the theme of this week's post!

Mini-Dab Nation, aka 6th Period AP Research.

Let's first look at my plan for my literature review. Last week, I outlined the subtopics into which my subtopic divides: Blood Diagnostics, Super-Hydrophilic Films, Analysis Techniques, Blood Properties, Substrates, and Key Illnesses/Conditions. These are the main sections I am envisioning for my literature review as well.

By using information about the limitations and consequences of current blood analysis techniques (e.g., hospital-acquired anemia from using large volumes of blood), I can establish the significance of my research with HemaDrop. Also, the subtopics are component parts of the research, which will still allow me to learn a lot and talk about pertinent information even though the research is pretty new and hasn't been explored yet. My plan is to investigate the viability of HemaDrop as a method for microliter blood analysis, but the information I find in my review of the literature on analysis techniques, substrates, and key illnesses/conditions will guide specifically what I will compare in my methods for my experiments.

When your list of goals triggers you hard.

Therefore, my literature review will serve the following purposes (inside the parentheses are the goals of a lit review about mini-dabbing): 

1. establish the limitations of current blood analysis techniques (basically the inadequacies of the current dab), 
2. demonstrate the need for and potential of microliter blood analysis (when you are in a confined space and are triggered), 
3. lay out the necessary criteria that a blood analysis technique must meet to be commercialized e,g., accuracy of current, large-volume methods (what will it take for mini-dabbing to go viral?),
4. explain the mechanisms of HemaDrop including super/hyper-hydrophilicity and phase separation (how the mini-dab mirrors the original dab), 
5. characterize the properties of blood when drying clotting in vitro compared to in vivo (I gave up trying to find parallels here)
6. find out normal/average human blood and canine blood composition  
7. determine pros/cons of various analysis techniques and find analysis techniques (vacuum-based) that HemaDrop samples can be used with to determine new information about blood composition (potentially via computer simulations first before testing),
8. and choose several key illnesses with specific markers in the blood for testing (methods could include comparative testing by adding markers to the blood).

This is also the order I will approach the subtopics in my lit review. One aspect of my literature review I have to be careful about is making sure my sources are in conversation, since the subtopics are all united seemingly only by HemaDrop. 

One specific source which I particularly enjoyed reading and which will definitely help me establish a gap in the research is "Senses, Sensors and Systems: A Journey Though the History of Laboratory Diagnosis" by Barthels and many other authors. "Senses, Sensors, and Systems" provides a thorough history of advances in laboratory diagnostics. I focused on the elements relevant to blood analysis.

First, the sections on blood lines and coagulation diagnostics explain the "cascade-and-arrest" mechanism of clotting of blood. Specifically, it explains all the proteins and triggering chemicals involved (fibrinogen, fibrin, thrombin, prothrombin, protein S, ionized Ca, etc.). The book even discusses the differences between in vivo (in the body) and in vitro (in lab settings) clotting. Next, it discusses near-patient diagnostics (point-of-care testing). For instance, it established the need for rapid identification of troponin concentration for heart attack patients. 

It even provided examples of physical concepts used in POCT. Finally, it discusses the requirements of all microtechnology that is POCT and stipulates the needs of blood diagnostics (use whole blood instead of just platelets or serum, yields quick results with high accuracy, uses small sample volumes, is simple to operate, has in-built quality control and minimal maintenance, and has an IT interface). Also, it cites a lot of articles in references about current methods of blood diagnostics which I had never heard of (e.g., dried blood spot chromatography), so I will look at those as well. Simply, the gap in the research is a device that fulfills those criteria.

Another exciting development is that my dad said he could arrange for me to meet up with/interview a pathologist and phlebotomist at his hospital to find out more about current blood analysis techniques. I'll keep you guys posted on that, as it becomes more concrete, but that information will be crucial for my literature review.

Finally, I'm all in for the John Oliver retort, as it would give us a chance to use our skills from Seminar and Research in a fun, engaging way. I'd be willing to work harder outside of school.

See you next week for some more great research!

Cheers,
YP

(836)