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Improvements in Methods for Detection of Properties of High Density Lipoprotein
Author(s) -
Yousefian Omid,
Hough Greg,
Hama Susan
Publication year - 2010
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.24.1_supplement.lb434
Subject(s) - enumeration , monocyte , chemotaxis , cell counting , high density lipoprotein , lipoprotein , medicine , chemistry , mathematics , cell , immunology , cholesterol , biochemistry , receptor , combinatorics , cell cycle
Objective We have developed an assay in which monocyte migration through filters, determines the level of inflammatory properties of low density lipoprotein (LDL) and high density lipoprotein (HDL) protective capacity. Automated monocyte enumeration demonstrated this property of HDL. Methods Cultured artery wall cells were exposed to either media alone, media plus LDL, or media plus LDL and HDL. The media from the treated cells was assayed in a monocyte chemotaxis assay. Image‐Pro Plus 5 software was used to automate the quantification of migrated monocytes. The accuracy of the data obtained from automated counting of images with 20X objective was compared with manual counting and tested by a two‐tailed paired t‐test. An integrated tool, Macro, was used to perform the automated enumeration on several images with 4X objective (the entire field). Summary of Results HDL, when presented to co‐cultures of artery wall cells together with LDL, significantly reduced the increased monocyte transmigration due to LDL alone. The statistical results, comparing the manual and automatic enumeration, demonstrated accuracy and precision (p=0.02, df=11, α= 0.05). Conclusion The results demonstrate that automated counting can replace manual counting, which should result in higher‐throughput of measurement of monocyte chemotaxis being used to determine the level of HDL proinflammatory or anti inflammatory properties.