Use of a Multiwell Assembly for Chemotaxis and Evaluation by Enzyme-Linked Immunosorbent Assay (ELISA)

Sorg, Clemens; Ramb, Christiane; Overwien, Bruno; Feige, Ulrich

doi:10.1016/s0171-2985(11)80030-6

Cited by 9 publications

(2 citation statements)

References 13 publications

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“…Although this technique provides an estimation of the relative leukocyte chemotactic activity of soluble substances, it is suggested that it gives partial and sometimes misleading information, and should be supplemented with other methods including visual assays [8]. Consequently, the Boyden chamber technique has been modified over the years [9], [10], [11]; however, none of these modifications allow the quantification of total migrating leukocytes concurrently with the determination of their phenotype.…”

Section: Introductionmentioning

confidence: 99%

Combined Boyden-Flow Cytometry Assay Improves Quantification and Provides Phenotypification of Leukocyte Chemotaxis

2011

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Chemotaxis has been studied by classical methods that measure chemotactic and random motility responses in vitro, but these methods do not evaluate the total number and phenotype of migrating leukocytes simultaneously. Our objective was to develop and validate a novel assay, combined Boyden-flow cytometry chemotaxis assay (CBFCA), for simultaneous quantification and phenotypification of migrating leukocytes. CBFCA exhibited several important advantages in comparison to the classic Boyden chemotaxis assay (CBCA): 1) improved precision (intra-assay coefficients of variation (CVs): CBFCA-4.7 and 4.8% vs. CBCA-30.1 and 17.3%; inter-observer CVs: CBFCA-3.6% vs. CBCA 30.1%); 2) increased recovery of cells, which increased assay to provide increased sensitivity; 3) high specificity for determining the phenotype of migrating/attracted leukocytes; and 4) reduced performance time (CBFCA 120 min vs. CBCA 265 min). Other advantages of CBFCA are: 5) robustness, 6) linearity, 7) eliminated requirement for albumin and, importantly, 8) enabled recovery of migrating leukocytes for subsequent studies. This latter feature is of great benefit in the study of migrating leukocyte subsets. We conclude that the CBFCA is a novel and improved technique for experiments focused on understanding leukocyte trafficking during the inflammatory response.

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Section: Introductionmentioning

confidence: 99%

Combined Boyden-Flow Cytometry Assay Improves Quantification and Provides Phenotypification of Leukocyte Chemotaxis

2011

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“…Abbreviations: fMLP, N-formyl-L-methionyl-L-ieucine-Lphenylalanine; LFA-1, leukocyte function associated antigen-1; PVP, polyvinylpyrrolidone. ing counting the cells under a microscope, or automatically using an Optomax image analyzer (M inkin et al, 1985), or using ELISA and radio labeling techniques (Gallin et al, 1973;Sorg et al, 1982;Sims et al, 1985).…”

Section: Introductionmentioning

confidence: 99%

Rapid densitometric determination of cell migration and cell adhesion in a microchemotaxis chamber

Klomp

Velde

Figdor

1989

Journal of Immunological Methods

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Chemotaxis: Basic aspects of methodology, mechanisms and pathology

Czarnetzki

Kownatzki

Dierich³

et al. 1983

Arch Dermatol Res

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This review is an attempt to discuss the basic conceptual tools that are a prerequisite for any clinical study of chemotactic defects. They include familiarity with definitions of common terms and with the merits and drawbacks of the several possible in vivo and in vitro assay methods. Cellular mechanisms involved in locomotion are complex. They include basic requirements for cell metabolism as well as receptor recognition, attachment to surfaces and contraction of the cytoskeleton of the cell. Of the many chemotactic factors reported, few are well characterized and universally agreed upon. Similarly, with the use of more stringent criteria, a number of clinical defects of chemotaxis have proven transitory or even artifactual.

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Use of a Multiwell Assembly for Chemotaxis and Evaluation by Enzyme-Linked Immunosorbent Assay (ELISA)

Cited by 9 publications

References 13 publications

Combined Boyden-Flow Cytometry Assay Improves Quantification and Provides Phenotypification of Leukocyte Chemotaxis

Combined Boyden-Flow Cytometry Assay Improves Quantification and Provides Phenotypification of Leukocyte Chemotaxis

Rapid densitometric determination of cell migration and cell adhesion in a microchemotaxis chamber

Chemotaxis: Basic aspects of methodology, mechanisms and pathology

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