Assessment of phagocytic activity of cultured macrophages using fluorescence microscopy and flow cytometry

Lokesh Sharma, Wenjun Wu, Sanjay L. Dholakiya, Samir Gorasiya, Jiao Wu, Ravikumar Sitapara, Vivek Patel, Mao Wang, Michelle Zur, Shloka Reddy, Nathan Siegelaub, Katrina Bamba, Frank A. Barile, Lin L. Mantell

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

Phagocytosis is the process by which phagocytes, including macrophages, neutrophils and monocytes, engulf and kill invading pathogens, remove foreign particles, and clear cell debris. Phagocytes and their ability to phagocytose are an important part of the innate immune system and are critical for homeostasis of the host. Impairment in phagocytosis has been associated with numerous diseases and disorders. Different cytokines have been shown to affect the phagocytic process. Cytokines including TNFα, IL-1β, GM-CSF, and TGF-β1 were found to promote phagocytosis, whereas high mobility group box-1 (HMGB1) inhibited the phagocytic function of macrophages. Here, we describe two commonly used methods to assess the phagocytic function of cultured macrophages, which can easily be applied to other phagocytes. Each method is based on the extent of engulfment of FITC-labeled latex minibeads by macrophages under different conditions. Phagocytic activity can be assessed either by counting individual cells using a fluorescence microscope or measuring fluorescence intensity using a flow cytometer.

Original languageEnglish (US)
Pages (from-to)137-145
Number of pages9
JournalMethods in Molecular Biology
Volume1172
DOIs
StatePublished - Jan 1 2014

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Fluorescence Microscopy
Phagocytosis
Flow Cytometry
Phagocytes
Macrophages
Fluorescence
Cytokines
Macrophage Colony-Stimulating Factor
Fluorescein-5-isothiocyanate
Latex
Granulocyte-Macrophage Colony-Stimulating Factor
Interleukin-1
Monocytes
Immune System
Neutrophils
Homeostasis

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Genetics

Cite this

Sharma, L., Wu, W., Dholakiya, S. L., Gorasiya, S., Wu, J., Sitapara, R., ... Mantell, L. L. (2014). Assessment of phagocytic activity of cultured macrophages using fluorescence microscopy and flow cytometry. Methods in Molecular Biology, 1172, 137-145. https://doi.org/10.1007/978-1-4939-0928-5_12

Assessment of phagocytic activity of cultured macrophages using fluorescence microscopy and flow cytometry. / Sharma, Lokesh; Wu, Wenjun; Dholakiya, Sanjay L.; Gorasiya, Samir; Wu, Jiao; Sitapara, Ravikumar; Patel, Vivek; Wang, Mao; Zur, Michelle; Reddy, Shloka; Siegelaub, Nathan; Bamba, Katrina; Barile, Frank A.; Mantell, Lin L.

In: Methods in Molecular Biology, Vol. 1172, 01.01.2014, p. 137-145.

Research output: Contribution to journalArticle

Sharma, L, Wu, W, Dholakiya, SL, Gorasiya, S, Wu, J, Sitapara, R, Patel, V, Wang, M, Zur, M, Reddy, S, Siegelaub, N, Bamba, K, Barile, FA & Mantell, LL 2014, 'Assessment of phagocytic activity of cultured macrophages using fluorescence microscopy and flow cytometry', Methods in Molecular Biology, vol. 1172, pp. 137-145. https://doi.org/10.1007/978-1-4939-0928-5_12
Sharma, Lokesh ; Wu, Wenjun ; Dholakiya, Sanjay L. ; Gorasiya, Samir ; Wu, Jiao ; Sitapara, Ravikumar ; Patel, Vivek ; Wang, Mao ; Zur, Michelle ; Reddy, Shloka ; Siegelaub, Nathan ; Bamba, Katrina ; Barile, Frank A. ; Mantell, Lin L. / Assessment of phagocytic activity of cultured macrophages using fluorescence microscopy and flow cytometry. In: Methods in Molecular Biology. 2014 ; Vol. 1172. pp. 137-145.
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