ASCLS Today Volume 33, Number 1

ASCLSToday Masthead 680

Volume 33, Number 1


Roslyn McQueen, PhD, CCRC, ASCLS President

As this is my first column of 2019, it is a good time to reflect on the past and plan for the future. In this month’s column, I will share my personal journey to becoming ASCLS president and discuss my plans for the rest of my presidential year.

I have been a continuous member of ASCLS for over 40 years, and I have never let my membership lapse. I have served on the local, state, regional, and national levels in various capacities from committee member, to committee chair, to state president, to Region IV director.

I was born Roslyn (no “a” in Roslyn) Wofford in Detroit, but have lived in Saginaw, Michigan, all my life. I have been married to Major General (Ret) Adolph McQueen for almost 40 years. I work at Hurley Medical Center in Flint and drive over 45 miles each way daily from Saginaw Township.

I attended Saginaw High School, graduating in the top 10 in a class of over 500. I received both my Bachelor of Science and doctorate from Michigan State University (MSU), College of Human Medicine, Department of Pathology. At that time, the Pathology Department was comprised of doctoral candidates from the College of Human Medicine, College of Osteopathic Medicine, and College of Veterinary Medicine. I was the only candidate from the College of Human Medicine.

My molecular research was performed in the laboratory of Dr. John Gerlach at MSU. Additionally, I received a Master of Arts from Central Michigan University, in Healthcare Administration.

First Laboratory Jobs
I was informed about the laboratory profession by my family physician but didn’t have any idea about what the profession entailed. Therefore, after my first term at MSU during the winter break, I went to every hospital in Saginaw and applied for a summer job in the laboratory. Of the three hospitals, only one was going to pay me, so I accepted a summer job in the laboratory at Saginaw General Hospital.

I was trained to perform the basic laboratory tests, starting with urinalysis. However, because I had a good work ethic and a strong desire to learn, the pathologist put me on a program to rotate to various departments whenever I returned from college. I was able to work in the lab during my summer and Christmas breaks and experience different departments. It was a very productive and enlightening experience.

St. Luke’s Hospital
After graduating from MSU, I was accepted into the Medical Technology Internship class at St. Luke’s Hospital in Saginaw. St. Luke’s was a very prestigious hospital where every technologist was ASCP certified. The medical technology program had the reputation of every student always passing the registry exam and scoring in the top 10 percent. I was fortunate enough to obtain the second highest grade in my class on the registry.

During my student year, I was introduced to the American Society for Medical Technology (which later became ASCLS) and the entire class became ASMT members. St. Luke’s interns were high achievers and very competitive. Our class participated in the Student Bowl during the Michigan Society for Medical Technology (later renamed ASCLS-MI) Annual Spring Meeting. Each member of the team was given a subject to focus on—mine was hematology/coagulation.

My team took first place in the Michigan Student Bowl and as state winners we were given a free trip to the ASCP national meeting in San Francisco. There, we competed against other teams on the national level and was fortunate enough to win the National Student Bowl. The Saginaw News wrote an article about our victory, naming our class the “Wiz Kids.”

Every tech (we referred to ourselves as “technologists” or “techs” back then, not medical laboratory scientists) at St. Luke’s was a member of the Saginaw Valley Society for Medical Technology (SVSMT). Our Education Coordinator/Lab Director Shirley Cresswell was an active member of the ASMT. She instilled in the class the fact that “medical technology was a profession, not a job,” therefore participation in our professional organization was mandatory. SVSMT was a very active society at the time. I served as treasurer, secretary, and eventually president of SVSMT.

Shirley Cresswell also encouraged me to participate in the Michigan Society for Medical Technology (MSMT) and invited me to attend board meetings with her. I started to attend MSMT Annual Meetings and eventually was recruited to serve on various annual meeting subcommittees. My first chairmanship appointment was workshop chair for the MSMT Spring Meeting. Later I was elected the MSMT District IX director, worked on several committees, became bylaws chair, and ultimately was elected MSMT president in 2002.

While serving as ASCLS Region IV director, Linda Gorman recommended me to serve on the ASCLS national Bylaws Committee. On the national level, I served on various committees, was appointed national bylaws chair, and eventually Linda Gorman suggested that I run for the position of Region IV director. While serving in my second term as Region IV director, Barbara Snyderman encouraged me to run for ASCLS president-elect.

I was hired at St. Luke’s hospital after graduation, worked as a staff tech, and was later promoted to Hematology Department supervisor. In this role, I worked with various heme-oncologists, residents, and other physicians. Later I was offered a job to work in research. Saginaw Cooperative Hospital was the Saginaw campus of MSU’s medical school where the university physicians established community campuses throughout the state. Ultimately, I left the clinical laboratory and, together with a colleague, started the Research Laboratory for Saginaw Cooperative Hospitals.

I have participated in various aspects of research starting with cancer research projects using mice assessing the efficacy of various drugs on the regression of tumor cells. This was performed under the direction of the heme-oncologist, who was director of internal medicine. Additionally, we performed in vitro susceptibility and synergism studies in infectious disease research under the direction of my boss, who was an infectious disease specialist and director of pediatric education.

In 1985 my boss, Dr. Ralph Gordon, was asked to assume the position of Director of Pediatric Education Residency Program at Hurley Medical Center, and he asked me to go with him and establish our research laboratory. There, I established the Pediatric Research and Education Laboratory. Through the years, we have submitted research papers that were published in leading scientific journals and made presentations at national, state, and local meetings. I conducted academic research at Hurley until 2000, when we changed focus to conducting industry-sponsored clinical trials.

Although, my primary focus is research, I have also provided teaching electives in infectious disease research and hematology to Hurley Medical Center pediatric residents and MSU medical students. I also serve as a hematology instructor for medical technology students at Hurley Medical Center and leukemia guest lecturer at Saginaw Valley State University. I have written chapters in hematology books for Bunny Rodak and J. Lynne Williams. I currently conduct clinical trials at Hurley Medical Center, as well as re-focus our efforts on academic research projects of molecular marker on red cells of patients with sickle cell disease.

I have been mentored by some of the most amazing individuals through the years from Michigan and throughout ASCLS. My education provided the initial skills and knowledge needed to begin my career, but ASCLS has provided me with the opportunities to become a professional. This mentorship has prepared me to assume the highest elected position in our illustrious organization. I do not take this lightly and am devoting every effort to fulfill the responsibilities associated with the office of president.

ASCEND - Exemplifying Sustainable Excellence in Laboratory Medicine
This year I have presented ASCEND as the platform for my 2018-19 presidential year, with the tagline, “Exemplifying Sustainable Excellence in Laboratory Medicine.”

ASCEND focuses on the changes that must occur as we meet the current challenges facing our organization, our profession, and our professionals. The letters in ASCEND stand for six key target areas.

A = Association Wellness
S = Sustaining the Strategic Map
C = Communication
E = Enhance Education
N = Networking
D = Diversity, and Leadership Development

Key platforms for this year include:

Constituent Society Wellness Taskforce. Under the Association Wellness platform, this new taskforce will take the data from the Root Cause Taskforce and establish a plan of action to support and strengthen our constituent societies.

ASCLS Annual Meeting – New Day, New Opportunities for Partnerships. An impending challenge facing ASCLS is the dissociation of our national meeting from the AACC Scientific Expo. Circumstances beyond our control resulted in our organization going out on our own, but this obstacle represents an opportunity for ASCLS to ASCEND to a new level. We look with optimism that this will open a new chapter in the life of our Society with the opportunity to sponsor an exceptional national meeting that will address the educational needs of our members.

This year’s meeting will be held in Charlotte, North Carolina, June 23-27. Our new partnership with the Association of Genetic Technologists (AGT) offers expanded and broadened education programming and access to a wider range of industry partners. We will have a vendor experience that will provide the face time our members seek with our industry partners. We encourage everyone to make plans to attend the new Annual Meeting. Also note that subsequent national meetings will be held in late June, instead of July. See you in Charlotte!

Communication Enhancements. This past fall we reinstituted Society News Now, our monthly e-newsletter, as one means of increasing communication with our membership. Susie Zanto established Society News Now to address the question of “What is ASCLS doing for me?” My goal is to continue her legacy and communicate activities that occur on a monthly or bi-monthly basis and provide information about what ASCLS is doing on behalf of our membership.

We have employed a director of marketing and communications and established a Marketing and Communications Committee. More information will be highlighted in future issues of ASCLS Today.

In 2019, ASCLS will continue to ASCEND to sustainable excellence through:
A: Working to identify and strength our struggling and/or inactive societies
S: Enhance the mission of our organization through our strategic plan
C: Increase communication
E: Enhance education
N: Increase our network within scientific associations
D: Provide leadership development for our diverse membership.

Roslyn McQueen is a research doctor at Hurley Medical Center in Flint, Michigan.


Courtney Bennetts, ASCLS Region IX Student Representative
Genaro Hernandez, Jr., MLT(ASCP), ASCLS Oregon Student Representative Region IX

ASCLS Region IX members gathered at the 2018 ASCLS Annual Meeting in Chicago.

ASCLS invests significant resources in student development to provide opportunities that can help them transition into highly qualified and successful professionals. This article details specific resources for students and shares how attending the ASCLS Annual Meeting, among other events, can help students develop professionally.

Developing Professional Membership Benefits
The ASCLS student membership is known as the Developing Professional membership. For a mere $24 (plus state dues) a student gets access to an enormous array of resources. Students are eligible for scholarships, travel grants, newsletters, discussion boards, student-centered activities, and a formal mentorship program.

Students can also volunteer at the local, state, and national level. By volunteering—even a little—students can begin to grow their support network, learn about opportunities (e.g., scholarships), and develop positive relationships with others who may provide a strong reference for a future application (e.g., a travel grant or job).

The best time to get involved is while we’re students, because we can follow in the footsteps of our current leaders to develop new innovative ideas on how to advocate for our field.

Genaro Hernandez, Jr., received the 2018 Diversity Advocacy Council Travel Grant and attended the 2018 ASCLS Annual Meeting in Chicago.

One Student’s Annual Meeting Experience
Genaro Hernandez, Jr., is a recent graduate and medical laboratory technician. He has volunteered in local and state ASCLS leadership positions, has attended several ASCLS meetings, and earned ASCLS scholarships and travel grants. He attended the 2018 ASCLS Annual Meeting in Chicago, and his experience there demonstrates the many benefits to students:

I am grateful to have attended the 2018 ASCLS Annual Meeting in Chicago. I truly enjoyed my experience and I hope to attend future Meetings. Here are some of my most memorable experiences from the meeting.

I met Daniel Haun, clinical instructor at Louisiana State University. He gave a fascinating presentation on virtual microscopy, which allows future medical laboratory techs to receive microscopy training completely online. Interestingly, instead of only providing a static image, techs are presented with a scannable image and context. Instructor Haun generates his own data using a customized microscope to collect images in the x, y, and z planes.

This was the first time I saw a medical laboratory tech create his own data. This was especially exciting to me because of my computer science background. I would like to work with medical laboratory data but most of the data is difficult to obtain. They are usually proprietary and/or contain personally identifiable information. Instructor Haun and I have been in contact since the Annual Meeting and he is helping me reconstruct his microscope, so I can generate my own images to work with. His presentation has encouraged me to use my interdisciplinary background and be attentive to potential informatic projects in my workplace.

Clarence Harris, head of international sales at US Bank, gave a powerful presentation on diversity and why it matters. Mr. Harris revealed that he was grateful for the opportunity to speak to us because he was indebted to the techs who provided STAT results when his wife was diagnosed with brain cancer. Those results were essential to her treatment.

He shared several important concepts. First, diversity without inclusion can be detrimental to an organization. Second, diversity and inclusion are essential for new ideas to surface. Third, those new ideas can help an organization to flourish. He noted that organizations must be open to change, or they will lose opportunities to remain viable. Mr. Harris provided real life examples of successful companies that followed these principles and unsuccessful companies that did not.

I was fortunate to receive a Diversity Advocacy Council (DAC) Travel Grant that allowed me to attend the Annual Meeting. I attended the DAC Meeting and listened while the council members earnestly discussed the delicate issue of how to manage and interpret transgender laboratory results. Attending the DAC Meeting prepared me to best serve transgender patients.

During the meeting, I asked my more experienced colleagues to recommend resources for developing leadership skills and attaining career goals. ASCLS Past President Debra Rodahl mentioned The 7 Habits of Highly Effective People, a book I have enjoyed since the meeting. Clarence Harris suggested learning from other successful people such as Zig Ziglar, Tom Hopkins, and Brian Tracy. These interactions have been helpful and have led me to explore additional related books and resources.

The afore mentioned are among my most memorable experiences from the 2018 ASCLS Annual Meeting—there are many more. I am proud to be a member of ASCLS and I am grateful for all the members who have supported my professional development. My goal is to remain active in ASCLS by continuing to serve in leadership roles. I look forward to the next Annual Meeting.

Opportunities for Students Close to Home
Students do not need to travel far to participate in ASCLS since local and state activities exist. These orchestrated events are crucial for networking and allow members of nearby communities to come together as one.

The future of our profession and its voice are the reasons we find it important to attend and volunteer. We want students to reach out, research, and find out when they can attend the events that have shaped so many of us into leaders. We, as developing professionals, want to advocate for students and motivate others to engage in our organization to bring laboratory professionals together on all levels for future generations.

Given the abundant opportunities for students to develop as professionals and leaders in ASCLS, we encourage all medical laboratory students to participate in the organization at any level they can. They will always find a knowledgeable long-term member willing to offer advice and support. If we can inspire and motivate students to take the step and pursue a position, then we have completed our mission.

Thank you, ASCLS!

Courtney Bennetts is a student in the University of Alaska Anchorage Clinical/Medical Laboratory Science/Research and Allied Professions Program in Anchorage. Genaro Hernandez, Jr., is a medical laboratory technician at Adventist Health in Portland, Oregon.



Floyd Josephat, EdD, MT(ASCP), ASCLS Education Scientific Assembly Chair

The CPA program is an advanced-practice program for the medical laboratory science profession with emphasis on the clinical aspects of laboratory medicine. Photo courtesy UAB.

Sam Cook mentioned about 50 years ago, “A Change is Gonna Come.” Laboratory medicine is also undergoing rapid changes driven by new technologies such as molecular and genomic testing. As new tests develop and become more complex, it is imperative that we prepare graduates to address issues they may encounter as a result of these changes.

Preparing students for the future should be our ultimate goal. This should involve a rigorous curriculum realignment or the addition of signature programs to address future clinical needs. The University of Alabama at Birmingham (UAB) Clinical Laboratory Science program has recognized there is a change coming and is addressing this pressing need by establishing a signature program to embrace the future of the clinical laboratory. The Clinical Laboratory Science program has developed a Clinical Pathologist Assistant (CPA) program to strategically prepare students for these changes.

Program Designed to Deliver Quality Patient Care
The CPA program is an advanced-practice program for the medical laboratory science profession with emphasis on the clinical aspects of laboratory medicine. Currently, professionally-relevant degree options are extremely limited for anyone wishing to continue their education once they have received their entry-level certification to work in an accredited laboratory. Additionally, the practice of laboratory medicine is sub-optimal due to inadequate collaboration among physicians, patients, nurses, and medical laboratory scientists where laboratory testing is concerned. Laboratory testing consists of many over- and under-utilized tests, leading to wasted money and resources and decreased quality of patient care.

Graduates of the CPA program would alleviate some of these communication problems by their ability to synthesize clinical and laboratory data and provide a narrative interpretation to help guide and support the decision-making process by the healthcare team, including patients, based upon medical evidence. The CPA program at UAB is also designed to prepare individuals with job skills directly related to one of the overarching mission pillars of UAB—“to deliver the highest quality patient care that reflects our ability to translate discoveries into revolutionary therapies in one of the nation’s largest academic medical centers.” This program will aid in improving the quality of patient care, as laboratory tests make up a large proportion of diagnostic tests used to guide patient care.

“It is estimated that more than seven billion clinical laboratory tests are performed in the United States each year, providing critical data that saves time, money, and lives by enabling early detection and prevention of disease,” said Peter G. Anderson, DVM, PhD, interim chair of the Department of Clinical and Diagnostic Sciences and professor of pathology at UAB Medicine. “We will prepare our students to serve as resources for clinicians and hospital staff, so they can choose wisely when ordering tests and interpreting the results. This will become especially important as more and more new tests become available.”

Employment Outlook
Employment opportunities for graduates of the CPA program initially will be expanded job responsibilities within their current organizations until the full scope of a clinical pathologist assistant job is defined and the job title becomes widely used. The College of American Pathologists, the leading organization of board-certified pathologists, has a current initiative to define clinical pathology job competencies for the CPA role. This proposed degree offers educational enhancement for current professionals that will enable them to perform advanced skill roles in the laboratory. For example, this may include serving on medical diagnostic teams with the anticipation of advancing into CPA jobs as they emerge.

These individuals will have the skill sets needed to assist the pathologist in his or her daily tasks including conducting test work-ups, interpreting data, providing consulting on appropriate test utilization and result interpretation, and other advanced practice job tasks. Graduates will be well prepared for these job tasks and consulting roles—to physicians, physician assistants, nurses, and other advanced practice providers—and as a result, will directly contribute to improving the quality of patient care from the critical aspect of laboratory testing.

Floyd Josephat is associate professor and program director for Clinical Laboratory Science and Clinical Pathologist Assistant Programs at the University of Alabama at Birmingham.


Franki-Marie Herdt, MLS(ASCP)CM, ASCLS Ascending Professionals Forum Region VIII Representative

I first stepped into a laboratory as a child in biology class, as most kids do. I loved the hands-on projects and real-world information we were given. Getting real answers to “Why is the sky blue?” was thrilling. As I got older, I continued to be drawn to the sciences. I took numerous science classes throughout high school—physics, biology, and zoology. I hadn’t specialized within a specific field of science until trying to decide on a college major.

Medical and crime shows directed me toward the type of science I know and love today. I decided on a degree in microbiology after going over all the different things I loved about my science classes—I really enjoyed sections of the classes that involved any microscopic work. After choosing microbiology, I started looking for a school that was close (but not too close) to home to pursue it further.

Surprisingly, this wasn’t as common of a field as I thought. After visiting a few local colleges, I decided on the University of Wyoming for my undergraduate degree in microbiology. Being a school deeply grounded in agriculture, my microbiology degree had a lot of veterinary background instead of medical. I believe this helped give me a well-rounded experience and knowledge base.

After graduating college, I had no idea what I wanted to do. I had worked at the State Veterinary Laboratory during most of my college career, so I was familiar with the research side and veterinary aspects of microbiology. I reviewed some of the classes I took my senior year hoping to narrow my search for my ideal job. One class that specifically caught my eye was Prion Biology. I was tasked with presenting on the Prion disease, Kuru, which helped me dive into medicine and the medical field. I slowly realized I wanted to go into more of the medical side of microbiology.

I applied at numerous hospitals around my area hoping to work in one of the laboratories. I discovered the laboratory I am currently at after many job searches online. I applied for one of the technologist positions, but I wasn’t qualified. Thankfully, since I had substantial laboratory background from the veterinary laboratory, they hired me as a lab assistant on the evening shift. I loved it! I got to work hands on with patient samples, including setting up samples in microbiology. However, after a while, I soon realized this wasn’t my forever job.

While working there, I noticed the work that the technologists were doing alongside me. They were analyzing hematology, chemistry, blood bank, and microbiology specimens and helping physicians make a diagnosis. I loved watching them work and eventually wanted a piece of the action. I wanted to see the organisms and know more about the diseases they were causing.

After getting to know my coworkers, I asked them how they got their technologist jobs. They told me about the certification process and the schools offering the programs. I knew I needed to find a program that allowed me to work while taking classes since I was already out of school and had bills to pay. I searched through many online programs that all had different requirements. Weber State University ended up being the right choice for me since it was an online program and clinicals were mixed with the courses instead of having it all at the end of the year.

While in school for the year, I did my clinicals concurrent with the classwork. This helped me get hands-on experience while learning the subject in that same week. I believe it helped me retain the knowledge a lot better since I was an online student. The clinicals also allowed me to get to know my laboratory better. I now knew what the technologists needed for testing and helped answer their questions before they asked them. It helped me be a better laboratory assistant for them.

I eventually was able to transfer into the Microbiology Department at my lab as a technologist and found my forever home. I absolutely love what I do, and my coworkers are just as passionate, which makes our jobs a lot more enjoyable. Who knew I would enjoy learning something new every day? Even though we work with specimens most people would find repulsive, I wouldn’t have it any other way. Getting to participate more with physicians and infection control gives me an excitement with which to come into work every day. I can see the difference we get to make for every single patient. The next hill to climb is to hopefully become the lead in my department.

There are many ways to get where I am today, and no way is better or worse than another. Some people benefit from an immersed program where others strive in a more freedom-based program. Once everyone starts to understand this and respect each other, the communications between inner-lab departments and the rest of the hospital will come together.

If I learned anything from this experience, it was that you will eventually find your forever home. When I was trying to figure out what I wanted to major in at college, my father told me to find something I love. The job will fall in place after that. I am proof of that advice. You don’t want to be stuck in a job you don’t love, so strive to find your perfect job and life. Realizing that every life experience is a stepping stone to something better can help to get through the boring classes, hard jobs, and unpleasant situations.

Franki-Marie Herdt is a medical technologist in microbiology at Cheyenne Regional Medical Center in Cheyenne, Wyoming.

Learn more about the ASCLS Developing Professionals Forum and the Ascending Professionals Forum.



Gerald D. Redwine, PhD, MT(ASCP), ASCLS Chemistry/Urinalysis Scientific Assembly Chair

Technology has taken cognition to the highest levels of learning in our labs, especially the technology for enzymes over the last three years on newly-acquired spectrophotometers. More than 30 years of using the faithful Gifford spectrophotometer slowly came to an end following impossible- or difficult-to-replace parts. But the replacement Lambda 265 photodiode array UV Vis spectrophotometer by PerkinElmer has more than made up for it. In vogue with today’s tech-savvy student, the technology adds new dimensions of learning and discovery; impossible in past chemistry labs. As a side note, it is also capable of doing certain molecular labs, but that is for other professors to discover.

Figure 1 Analyzer and laptop, comparable L x W, and weight.
Figure 2 Real-time measurements at 15-second internals.

Following the initial setup, the need for support was nonexistent. Working with the laptop computer program and the small footprint analyzer (Figure 1) is relatively simple. We added other accessories such as the flow cell with a peristaltic pump (not shown) that sets between the laptop and analyzer. We also added a “Water Jacketed Single Cell Holder,” located between the horseshoe-shaped analyzer, and its temperature regulated “Peltier Controlled Fluid Circulator,” seen to the right of the analyzer. The analyzer is truly an open system, not only for reagents but regarding its open spectral analysis, worry-free of stray light due to its high energy xenon flash lamp.

Altogether it means that for the first time students can see, in real time, endpoint and kinetic reactions as the computer presents or graphically tracks the reactions at programmed intervals of choice to completion (Figures 2, 3). This ability accentuated learning beyond recall and comprehension to the higher level of application in Bloom’s taxonomy.

The days of recording point-by-point enzymatic activity on graph paper are over because students can right click, copy, and paste any or all resulting data or graphics into one or several media like Excel (Table 1), Word, or PDF for future reference. Students also can simply click “Export,” with some selectivity to similar media.

Even more exciting, students in the included example observed in real time the enzymatic activity. They zoomed in during and following the kinetic reaction and witnessed the positive (increasing) variance in the alkaline phosphatase (AP/ALP) activity while remaining compliant with the manufacturer’s test requirement (Figure 2). Cognitive discussions ensued concerning the abstract concept of Lambert-Beer’s Law and the resulting equation (Absorbance = 2-log%T)1,2 relating absorbency to transmitted light.

The ability to export data into an Excel file (Table 1) made the analysis a snap and facilitated transitioning to even higher levels of cognition. They selected the most stable minute from Figure 2, compared it to values in Table 1, and completed their calculation to quantitate the AP activity.

Table 1 Exampled Alkaline Phosphatase (AP)

Time (Sec) 405.000nm ΔA ΔA/min
0 0.574    
15 0.581 0.007  
30 0.586 0.005  
45 0.593 0.007  
60 0.599 0.006 0.025
75 0.608 0.009  
90 0.611 0.003  
105 0.620 0.009  
120 0.626 0.006 0.028
135 0.631 0.005  
150 0.638 0.007  
165 0.646 0.008  
180 0.654 0.008 0.028


U/L=  ΔA/min × 1000 × 1000 × tf × tvr
             Molar Absorptivity x lp × sv

  • ΔA/min – change of absorptivity/min
  • Molar Absorptivity for Alkaline Phosphatase, 18,880
  • 1000 – converts mL to a liter
  • 1000 – converts millimoles to micromoles
  • tf – temperature factor, usually 1.00
  • trv – total reaction volume
  • lp – light path in centimeters, 1.0
  • sv – sample volume in mL

Results in SI Units/Liter (U/L):
U/L= 0.028 × 1000 × 1000 × 1 × 1.02 mL = 76 U/L (Ref.30 -165)
                         18,880 x 0.02 mL

Figure 3. Extended real-time measurements at 15-second internals

In the past, manually graphing point-to-point reactions was the only choice without the real-time monitoring capability, which limited the desired learning outcome to a calculated result. However, with technology, although the goal of getting a result from the analysis was complete, learning continued.

Reflecting on the meaning of all they had accomplished catapulted the students to the next higher level of cognition in Bloom’s taxonomy, synthesis, by predicting that similar reactions would occur with other enzymatic reactions. With assistance, students then moved into the highest level of cognition, evaluation. They focused on the AP measurement frequency of 405 nm in an experiment, then the spectral evaluation at two points. The beginning and the end of the analysis, with an absorptivity of 0.574, and 0.654 respectively, as indicated in Table 1, facilitated the learning.

Admittedly, students needed guidance in expanding their knowledge into the realm of evaluation, accomplished with a “what if” experiment. Testing to see what extending the number of minutes of testing (Figure 3) would accomplish succeeded in reaching the highest level of cognition, evaluation. So, what did they discover? Three things quickly became obvious, which stimulated other learning investigations.

First, they found that a relatively small quantity of patient AP enzymes in the zero order (excess substrate) reaction did not exhaust it, as seen by the continual increase of product. Secondly, the longer the reaction, the greater the instability. Thirdly, the longer the reaction, the less predictable the rate of production. The visual results also begged for a future investigation of the lag phase by starting the measurement shortly after mixing the patient sample with the substrate.

Figure 4. Left slightly-upper 3D view, full spectrum scan, 405 nm indicated.
Figure 5. Top 3D view, full spectrum scan, 405 nm indicated.
Figure 6. Left lower 3D view, full spectrum scan, 405 nm indicated.
Figure 7. Full spectrum scan truncated at 900 nm, 405 nm indicated.
Figure 8. Subtle changes measurable differences.

In the meantime, the ability for students to graphically observe the enzymatic reactions in real time helped facilitate learning at the highest level of cognition and encouraged them to delve into research of spectral analysis; in short, it facilitated critical thinking. Students discovered that the spectrum was examinable not only in 3D but any angle could be examined, each with their advantage. For example, Figure 4 revealed to the students that absorbances beyond 2A were possible and had a very similar pattern throughout the spectrum from 200 nm to 1100 nm. That is, most of the activity was between 200 nm and slightly more than 400 nm. Also, how and why the manufacturer decided to use 405 nm as the point of measurements became a point of interest.

From the top (Figure 5) the spectral scan reveals a similar pattern over time. That is, some pre-activity, followed by heavy activity, and resolve to post activity. Nevertheless, similar results came from the products beginning with the initial timing and continued throughout the three minutes. Surprisingly, perhaps the best view was the lower view (not bottom) in Figure 6 that revealed several interesting facts including the reason for selecting the wavelength at 405 nm for measurement.

The first note of interest is the increasingly larger negative values and increasingly larger negative spikes in the UV range over time. A careful study of these negative values, especially the spikes, seems to coincide with the positive varianceand peaks seen in Figure 3. This coincidence indicates that enzymatic activity at the measured frequency is mirrored negatively in the UV range. Interesting.

The next notable point is the color code scheme in a 3D object with divisions based upon absorptivity. Having two shades of green (0A-2A) seemingly representing go or good; yellow and orange (2A-3.5A), representing caution or warm; red (3.5A-4A) representing stop or hot; and two shades of blue (0A- -2.5A) in the UV range representing cold and negative. The darker green is between 0A and 1A and is the range used for our analysis. But why? This question brings us to the most informative presentation from this (Figure 4) vantage point. At the indicated 405 nm frequency there is a smooth curve representing a dependable change in absorbance (ΔA) ideal for the measurement of AP enzymatic activity. However, that is not the complete answer; that requires the focus on the frequency of 405 nm, resulting in other ways technology makes the study of enzymes cognitively stimulating.

With a look at the 3D models in our rearview mirror, a more informative look at the normal spectral scan is possible. With a look at the indicated 405 nm arrow in Figure 7, perhaps a bit of reflection helps us to recall that activities here, as seen in Figure 3, are mimicked in the UV range, as seen in Figure 6 and now Figure 7. We also see that before the negative activity in the UV range there is a positive activity, which is a point of interest begging an investigation; but that is beyond the scope of this report. Nevertheless, at this point, we have more questions than answers for reasons the manufacturer chose 405 nm as indicated by the arrow. Why not choose 400 nm? It seemed just as good. Or better yet, why not choose a range at the peak that is before 400 nm? The answer to the last question is more directly answerable.

We note that the peak (not before seen) is around 2A or 0.1 to 1%T (transmittance) using the Lambert-Beer’s equation (Absorbance = 2-log%T) for determining absorptivity, and subtle differences could result in large errors. Therefore, our conclusion correlates with research literature stating that small changes at low %T relates to large changes in absorbances and creates greater opportunity for error.1 Even more perilous is a sharp peak that would produce less- or impossibly-reliable results. Therefore, based on our results, greater than 1%T and less than 100%T, usually approaching 1A is desirable in the measurement of the AP enzyme and coincide with medical laboratory science measurements in general. More succinct research shows that 36.8%T or 0.434A presents the least error, but between .1A and .7A or 20%T to 80%T the error is minimum.1

The mystery is finally solved by zooming in on the area of interest, the 405 nm wavelength chosen by the manufacturer and enlarged in Figure 8. Depicted in this figure are the initial reading (purple) and the final reading at 180 seconds (red) at the estimated 405 nm (green) indications. Close examination revealed that the slope moves slightly to the right over time, which brings the absorbance reading increasingly towards an absorbance of one (1A). This view further explains why the manufacturer chose the 400 nm (black dots). The slope of the line is too steep, which would cause more difficulty in making a distinction between absorptivity. Finally, seen is the 405 nm is just before the reduced enzymatic activity.

One last thought, it is known that the accuracy of absorbance is dependent upon a ratio of the natural bandwidth to the spectral bandwidth of less than 10% to produce 99.5% accuracy.1 Therefore, we are led to believe that the diode array of the Lambda 265 is so discerning on an otherwise forebodingly steep slope that it can repetitively produce accurate results. So far that has proven to be the case.

One could ask other questions, such as, how could the manufacturer pinpoint this precise location without technology, or did they have similar technology? One may also speculate that a study of the lag phase as suggested earlier would produce a similar pattern, or would it? Is the background noise in the UV range (190nm~380nm)3 the result of the enzymatic activity on the substrate, or does the substrate cause it? Doing this experiment would prove our speculative statements made concerning the relationships of the positive peaks of activity seen in Figure 3 and the negative ones in Figure 6 in the UV range.

With the technology of the Lambda 265 analyzer, answering these questions are made easy by monitoring the spectrum prior, during, and following the addition of the patient sample containing the alkaline phosphatase (AP). Additional questions are answerable through this approach, such as, “What is the spectral pattern while the reagent is reaching optimum temperature, also, what about the patient sample while it does the same when mixed into the substrate?” Why should students not answer these questions for themselves, rather than relying upon others for the answers? It is called research, brought about by curiosity, aiding the students in the highest levels of cognition—analysis and evaluation—easily accomplished without endangerment with the aid of technology.


  1. Burtis CAA, Edward R.; Bruns, David E. Tietz Fundamentals of Clinical Chemistry. 6 ed 2008.
  2. Bishop ML, Fody, Edward P.; Schoeff, Larry E. Clinical Chemistry: Principles, Techniques, and Correlations 7ed: Lippincott Williams & Wilkins, a Wolters Kluwer Business; 2013.
  3. Sciences PLaA. Lambda 265 Users Guide. In: Service LSa, ed. Shelton, CT: PerkinElmer:8.

Gerald Redwine is associate professor at Texas State University Clinical Laboratory Science program in San Marcos, Texas.