In Vitro Phagocytosis Assay: A Live Imaging Based Method to Study Real-time Astrocyte Mediated Synaptosome Phagocytosis
Published: April 30, 2023
Abstract
Source: Byun, Y. G., Chung, W. S. A Novel In Vitro Live-imaging Assay of Astrocyte-mediated Phagocytosis Using pH Indicator-conjugated Synaptosomes. J. Vis. Exp. (2018)
In this video, we evaluate the phagocytic capacity of purified astrocytes in vitro by live imaging assay. The assay serves as an effective screening platform to identify chemical compounds that can enhance or inhibit the phagocytic capacity of astrocytes.
Protocol
1. Phagocytosis Live-imaging Assay (Figure 1) Prepare a 24-well plate (or any plate/dish that is prepared for live-imaging) in which astrocytes are confluent (generally, 7 to 10 days of incubation after the purification is ideal for stabilizing the cells). Remove the media in each well and wash the wells with 1 mL of Dulbecco’s phosphate buffer saline (DPBS), three times. To minimize air exposure, wash the wells quickly. Add 300 µL of immunopann…
Representative Results
Figure 1. Schematic of phagocytosis live-imaging assay.
Divulgations
The authors have nothing to disclose.
Materials
| pHrodo red, succinimidyl ester | Molecula probes | P36600 | For pH indicator conjugation |
| Fetal bovine serum (FBS) | Gibco | 16000-044 | |
| DMEM | Gibco | 11960-044 | |
| dPBS | Welgene | LB001-02 | |
| Phagocytosis live imaging assay | |||
| Juli stage | NanoEntek | ||
| Time Series Analyzer V3 plugins | https://imagej.nih.gov/ij/plugins/ time-series.html |
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Citer Cet Article
In Vitro Phagocytosis Assay: A Live Imaging Based Method to Study Real-time Astrocyte Mediated Synaptosome Phagocytosis. J. Vis. Exp. (Pending Publication), e20694, doi: (2023).