Immunological tools: Engaging students in the use and analysis of flow cytometry and enzyme‐linked immunosorbent assay (ELISA)

Abstract

Flow cytometry and enzyme-linked immunosorbent assay (ELISA) are commonly used techniques associated with clinical and research applications within the immunology and medical fields. The use of these techniques is becoming increasingly valuable in many life science and engineering disciplines as well. Herein, we report the development and evaluation of a novel half-semester course that focused on introducing undergraduate and graduate students to advance conceptual and technical skills associated with flow cytometry and ELISA, with emphasis on applications, experimental design, and data analysis. This course was offered in the North Carolina State University Biotechnology Program over three semesters and consisted of weekly lectures and laboratories. Students performed and/or analyzed flow cytometry and ELISA in three separate laboratory exercises:(1) identification of transgenic zebrafish hematopoietic cells, (2) analysis of transfection efficiency, and (3) analysis of cytokine production upon lipopolysaccharide stimulation. Student learning outcomes were achieved as demonstrated by multiple means of assessment, including three laboratory reports, a data analysis laboratory practicum, and a cumulative final exam. Further, anonymous student selfassessment revealed increased student confidence in the knowledge and skill sets defined in the learning outcomes.

Evaluation of99mTc-HYNIC-VCAM-1scFvas a Potential Qualitative and Semiquantitative Probe Targeting Various Tumors

“…There are various methods to detect protein expression using molecular biological techniques [ 21 ], such as immunohistochemistry, immunofluorescence, western blotting, and ELISA. Among them, ELISA is mainly applied for the detection of secretory proteins [ 22 ]. Although the specificity of western blotting is high, the procedure is complex.…”

Acetic acid alleviates the inflammatory response and liver injury in septic mice by increasing the expression of TRIM40

“…no. BGK06804) levels in ascites were detected using ELISA kits (PeproTech, Inc., Rocky Hill, NJ, USA) as previously reported (32). The values are recorded as ng/mg of protein.…”

A full semester flow cytometry course improves graduate and undergraduate student confidence

“…Ott et al report about a course with an in‐depth study of immunological tools, including half of a semester focused on flow cytometry. By delving deeper into flow cytometry, the students were able to develop expertise and were exposed to numerous applications . The undergraduate students reported increases in confidence across the board and had high satisfaction with the half semester flow cytometry course.…”

“…However, a solid understanding of the physical technology, the biological principles, and the data processing is necessary for students to effectively gather data with a flow cytometer . One previous publication detailing the effectiveness of an 8 week course cites student feedback that due to the complexity of the topic they wanted the course to be an entire semester …”