Archived Abstracts
Using Image J to Evaluate Cell Number in Cultured RAW 264.7 Macrophages
Location
Room 232, Schewel Hall
Access Type
Campus Access Only
Entry Number
72
Start Date
4-8-2020 1:30 PM
End Date
4-8-2020 1:45 PM
Department
Biology
Abstract
A comparison of the response of RAW 264.7 cells to bone marrow derived macrophages (BMDM) stimulated with bacterial lipopolysaccharide O55:B5 from E. coli (LPS) intended to determine if the cell line and primary cells demonstrated equivalent performance in nitric oxide production. Differential growth rates inhibited direct comparisons. The purpose of this study is to count the macrophage cells on a 24-well plate using the software ImageJ. ImageJ is a processing system used to count objects in images. We use ImageJ to count macrophage cells by uploading pictures of macrophages taken during experiments. Images of higher microscopic magnifications allowed for more accurate cell counts. Images under lower magnifications were too dense to count. At lower magnifications (200X) the software could not differentiate individual cells causing clusters to be counted as one large cell. Pictures with higher magnifications were differentiated. After counting cells, the total amount of cells in the 24-well plate was calculated. The diameter of the well was used to make these calculations. Comparison of RAW 264.7 cells to BMDM allows for effective assessment of RAW 264.7 cells as a substitute for BMDM and led to a reduction in the need for animals to study macrophage function.
Faculty Mentor(s)
Dr. David O. Freier
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Using Image J to Evaluate Cell Number in Cultured RAW 264.7 Macrophages
Room 232, Schewel Hall
A comparison of the response of RAW 264.7 cells to bone marrow derived macrophages (BMDM) stimulated with bacterial lipopolysaccharide O55:B5 from E. coli (LPS) intended to determine if the cell line and primary cells demonstrated equivalent performance in nitric oxide production. Differential growth rates inhibited direct comparisons. The purpose of this study is to count the macrophage cells on a 24-well plate using the software ImageJ. ImageJ is a processing system used to count objects in images. We use ImageJ to count macrophage cells by uploading pictures of macrophages taken during experiments. Images of higher microscopic magnifications allowed for more accurate cell counts. Images under lower magnifications were too dense to count. At lower magnifications (200X) the software could not differentiate individual cells causing clusters to be counted as one large cell. Pictures with higher magnifications were differentiated. After counting cells, the total amount of cells in the 24-well plate was calculated. The diameter of the well was used to make these calculations. Comparison of RAW 264.7 cells to BMDM allows for effective assessment of RAW 264.7 cells as a substitute for BMDM and led to a reduction in the need for animals to study macrophage function.