OCR Text |
Show 61 college of pharmacy Fluorescence microscopy is a commonly used technique to assess cell viability in response to vari-ous environmental conditions, including drug cytotoxicity. In most methods, dye-marked cells are counted under fluorescence microscopy, yielding an accurate assay, but a time-intensive process requiring multiple fields to be counted manually. This precludes high-throughput analysis for mul-tiple screens. Alternatively, flow cytometry can be employed to determine cell viability quickly and effectively; however, methods development for flow cytometry can be extensive, requiring validation of new methods to properly stain and count cells. Additionally, substantial time and training must be invested to ensure proper instrument settings, such as flow rate and compensation, adjusted for each stain and cell type. Alternatively, the most common cell counting method uses a haemocytometer as a quick, direct way to count cells by eye and determine percent viability with the aid of a trypan blue stain. This method, however, tests only a small volume of the total culture (usually ~1-10%) prompting questions of how accurately haemocytometers measure cell viability in toxicity testing. This research aims to compare three common cell counting methods for assessing viability (fluorescence microsco-py, flow cytometry, and haemocytometry) to determine method accuracy and efficiency for measuring cytotoxicity of macrophages in response to antibiotic dosing. Preliminary experimentation indicates that the development of application-specific cell staining protocols is a crucial first step in assessing high-throughput cell counting methodologies. COMPARISON OF COMMON CELL-COUNTING METHODS FOR ACCURATE ASSESSMENT OF ANTIBIOTIC-INDUCED CYTOTOXICITY IN MACROPHAGES Sherry Davidoff, Dolly Holt, Benjamin Brooks, (Amanda Brooks, David Grainger) Department of Bioengineering Department of Pharmaceutics and Pharmaceutical Chemistry University of Utah UNDERGRADUATE RESEARCH ABSTRACTS Sherry Davidoff Dolly Holt Benjamin Brooks Amanda Brooks David Grainger |