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Phagocytosis

Phagocytosis is the process by which cells engulf and internalize solid particles, macromolecules, or other matter from their surrounding environment.
This cellular process is crucial for immune function, tissue homeostasis, and pathogen clearance.
Phagocytes, such as macrophages and neutrophils, play a key role in phagocytosis, engulfing and digesting foreign substances, cellular debris, and infectious agents.
Studying the mechanisms and regulation of phagocytosis is essential for understanding immune responses, inflammation, and the pathogenesis of various diseases.
Advances in research tools and techniques, like PubCompare.ai, can help identify the most reproducible and accurate phagocytosis protocols, boosting productivity and accelerating scientific discoveries in this important area of cell biology.

Most cited protocols related to «Phagocytosis»

1
Caspase-1 Regulates Alzheimer's Pathology
Cited 27 times
Caspase-1 activation of human and mouse brain tissue were analyzed by Western blot of cleaved caspase-1. IL-1β was quantified by ELISA. Microglial ASC speck formation was detected by immunohistochemistry. All mice were on C57/Bl6 background, including WT, NLRP3−/−,27 (link), APP/PS15 (link), APP/PS1/NLRP3−/−, Caspase-1−/−,28 (link), APP/PS1/Caspase-1−/− and were analyzed for cognitive function using the Morris Water Maze, the object recognition test and open field behavioural testing. Synaptic plasticity was determined by measuring long term potentiation (LTP) in acutely isolated hippocampal slices. Spine density was assessed by analyzing mid apical dendritic sections of pyramidal CA1 neurons. Cerebral Aβ load was determined by thioflavin-S-histochemistry of serial sections. Sequential extraction of homogenized brains by radio-immunoprecipitation assay, sodium dodecyl sulfate buffer and formic acid was employed to determine Aβ levels. Aβ nitration was determined by ELISA and immunohistochemistry using a specific antibodies against 3NTyr10 (link)-Aβ25 (link). Western blot detection was used to analyze the protein levels of APP, CTFs, Aβ, BACE1, IDE and NOS2. Inflammasome activation was confirmed by detection of ASC speck formation in microglia isolated from adult mouse. Microglial Aβ phagocytosis was determined after peripheral injection of methoxy-XO4, isolation of microglia and subsequent FACS analysis. Confirmatory immunocytochemistry was performed using antibody IC16 and the lysosomal marker LAMP2. Plaque morphology and microglial Aβ uptake was analyzed by coimmunostaining with Iba-1, methoxy-XO4 and IC16. mRNA levels of IDE, NEP, M1 and M2 markers were determined either from sorted microglia or from brain tissue by qPCR.
Heneka M.T., Kummer M.P., Stutz A., Delekate A., Schwartz S., Saecker A., Griep A., Axt D., Remus A., Tzeng T.C., Gelpi E., Halle A., Korte M., Latz E, & Golenbock D. (2012). NLRP3 is activated in Alzheimer´s disease and contributes to pathology in APP/PS1 mice. Nature, 493(7434), 674-678.
Publication 2012
Adult Antibodies BACE1 protein, human Biological Assay Brain Buffers Caspase 1 Cognition Dendrites Enzyme-Linked Immunosorbent Assay formic acid Histocytochemistry Homo sapiens Immunocytochemistry Immunoglobulins Immunohistochemistry Immunoprecipitation Inflammasomes Interleukin-1 beta isolation LAMP2 protein, human Long-Term Potentiation Lysosomes Mice, Laboratory Microglia Morris Water Maze Test Neuronal Plasticity Nitrates Nitric Oxide Synthase Type II Phagocytosis Proteins Pyramidal Cells RNA, Messenger Senile Plaques Sulfate, Sodium Dodecyl thioflavine Tissues Vertebral Column Western Blotting
2
Quantifying Phagocytic Bead Internalization
Cited 12 times
To ensure that phagocytosis measured by flow cytometry resulted in actual bead uptake, and not bead attachment alone, we performed imaging flow cytometry to assess the amount of bead attachment and internalization. Immune complexes were formed with HIVIG, IVIG and no antibody control as described above. WBCs were incubated with immune complexes at 4 °C and 37 °C for 1 h. Cells from 12 wells were then pooled, washed and stained with CD66b-AF647 (Biolegend) and DAPI (Invitrogen). Bead attachment and internalization by CD66b+ cells were visualized using an ImageStreamX MkII (EMD Millipore) across 500,000 independent neutrophil events (60× objective, 405, 488 and 642 nm lasers, with extended depth of focus) and quantified using IDEAS 6 software utilizing the internalization module. An internalization feature was employed describing the ratio of fluorescence intensity of the beads inside the cell against whole cell intensity, with positive scores representing greater internalization.
Karsten C.B., Mehta N., Shin S.A., Diefenbach T.J., Slein M.D., Karpinski W., Irvine E.B., Broge T., Suscovich T.J, & Alter G. (2019). A versatile high-throughput assay to characterize antibody-mediated neutrophil phagocytosis. Journal of Immunological Methods, 471, 46-56.
Publication 2019
Alexa Fluor 647 CEACAM8 protein, human Cells Complex, Immune DAPI Flow Cytometry Fluorescence HIV hyperimmune globulin Immunoglobulins Intravenous Immunoglobulins Leukocytes Neutrophil Phagocytosis
3
Statistical Analysis of iPSC-RPE Patch Validation
Cited 11 times
Statistical analysis for in vitro functional validation of iPSC-RPE-patch was performed using R-software and where applicable the Dunn.test package. Data was first assessed for normality by determining data skewness, kurtosis, and q-q plots. All gene expression, TER, shape metrics, and phagocytosis data were found to have skewness or kurtosis values outside of a −1 to 1 range and showed significant deviance in q-q plots and thus were treated as non-normal. Dunn’s test was therefore used for reporting multiple pairwise comparisons after a Kruskal-Wallis test for stochastic dominance among k groups was performed. A Bonferroni-Dunn correction was used for all pair-wise comparisons and an adjusted p-value < 0.05 (*), 0.01 (**), and 0.001 (***) were considered significance. For flow cytometry data in figure 1D, quartile regression models and an ANOVA were utilized to determine differences in slope from 0. For principle component analysis data from phagocytosis, TER, and gene expression profiles across all days and clones was scaled from 0 to 1 using the total pooled data for each metric. PCA was performed and clustering was shown based on k-nearest neighbors. Bootstrap hierarcial cluster of PCA was performed to show similarity between different iRPE samples from three donors (*p<0.05). A linear mixed model (LME) and ANOVA was performed to determine if statistical differences between different transplant groups in pigs. The equation function used in the MATLAB fitlme function was: Data ~ Week + Group*Component + (1|Pig_Name). A linear regression of the AUC mfERG waveform component was also performed in Graphpad and the elevation and y-intercepts determined. A p-value of 0.05 or less was determined to be statistical significant for both the LME and the linear regression. ANOVA was used to perform statistics on OKN data, *p<0.05, **p<0.01 were considered significant. To count the number of nuclei (DAPI) in the implant region (either empty scaffold or iRPE-Patch) in pigs was normalized to the corresponding healthy region on the same section. The same distance across the retina (~ 400 μm) was used for the implant area and the corresponding healthy area to count. In RCS rats a ~ 500 μm region was used for counting the number of nuclei in the ONL. An unpaired two-tailed t-test was performed for TUNEL positive nuclei and the number of nuclei in ONL of RCS and pigs, and resulted in a significant difference between the groups (** p<0.01, and ***p<0.001).
Sharma R., Khristov V., Rising A., Jha B.S., Dejene R., Hotaling N., Li Y., Stoddard J., Stankewicz C., Wan Q., Zhang C., Campos M.M., Miyagishima K.J., McGaughey D., Villasmil R., Mattapallil M., Stanzel B., Qian H., Wong W., Chase L., Charles S., McGill T., Miller S., Maminishkis A., Amaral J, & Bharti K. (2019). Clinical-Grade Stem Cell-Derived Retinal Pigment Epithelium Patch Rescues Retinal Degeneration in Rodents and Pigs. Science translational medicine, 11(475), eaat5580.
Publication 2019
Cell Nucleus Clone Cells DAPI Donors Flow Cytometry Gene Expression Grafts Induced Pluripotent Stem Cells In Situ Nick-End Labeling neuro-oncological ventral antigen 2, human Phagocytosis Pigs Rattus norvegicus Retina
4
Phagocytosis Assay of K. pneumoniae
Cited 10 times
The phagocytosis assay of K. pneumoniae strains was performed by an established method (23 (link)). An inoculum of 107 CFU was mixed with 106 freshly isolated human neutrophils or mouse macrophage cell line RAW-264.7 in RPMI 1640 media (23 (link)). After 15 min of incubation, extracellular bacteria were washed out, and 100 μg/ml gentamicin was added to kill any remaining extracellular bacteria. Cell-associated bacteria were quantified at zero and 30 min after the end of incubation by plating on LB agar plates after lysis of the cells with 0.1% Triton X-100. Initial experiments established that the magA knockout mutants were no more sensitive to the detergent than the wild type. To determine whether cell-associated bacteria counted by the aforementioned method were indeed ingested by human neutrophils, fluorescence microscopy using confocal microscope was performed. Before fluorescence microscopy, plasmid GFPuv (CLONTECH Laboratories Inc.), which carries a gene that encodes green fluorescent protein, was electroporated into selected K. pneumoniae strains.
Fang C.T., Chuang Y.P., Shun C.T., Chang S.C, & Wang J.T. (2004). A Novel Virulence Gene in Klebsiella pneumoniae Strains Causing Primary Liver Abscess and Septic Metastatic Complications. The Journal of Experimental Medicine, 199(5), 697-705.
Publication 2004
Agar Bacteria Biological Assay Cells Detergents Genes Gentamicin Green Fluorescent Proteins Homo sapiens Klebsiella pneumoniae Macrophage Microscopy, Confocal Microscopy, Fluorescence Mus Neutrophil Phagocytosis Plasmids RAW 264.7 Cells Strains Triton X-100
5
Neutrophil Functional Assays Compendium
Cited 9 times

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Publication 2012
Adoptive Transfer Bacteria Biological Assay Cells Chemiluminescence Chemotaxis Differentiations, Cell F-Actin GLRX protein, human HL-60 Cells isolation Lentivirus Luminol Neutrophil Peritonitis Phagocytosis Phalloidine RNA, Small Interfering Stain, Giemsa Superoxides

Most recents protocols related to «Phagocytosis»

1
Microglial Phagocytosis in Neurodegeneration
Not available on PMC !

Example 7

WT or STAT1−/− microglia cells were incubated with fluorescent beads to test their phagocytotic ability. To examine the effect of secreted factors in medium, both cells were kept in medium from WT cells or in medium from STAT1−/− cells. Imaging and quantification showed phagocytosis was similar in all conditions tested (FIGS. 17A and 17B). We also measured mRNA of SV2a and CCR2, key genes involved in phagocytosis process. Their expression levels were similar between APP/PS1 and APP/PS1/STAT1−/− (FIG. 17C and FIG. 17D).

US11873321B2. Compositions and methods for treating alzheimer's disease (2024-01-16). Generos Biopharma Ltd. [CN]. Inventors: Xin-Yuan Fu [US], Yi Zhou [SG].
Patent 2024
Cells Genes Microglia Phagocytosis RNA, Messenger STAT1 protein, human
2
Quantifying Microglial Aβ Phagocytosis
Microglial phagocytosis was evaluated by quantifying the degree of internalization of the fluorescent HiLyte Fluor 488-labeled Aβ1-42 peptide (AnaSpec, AS-60479-01). Cells were seeded in 8-well LabTek removable chamber slides. After treatment, 1 μM HiLyte Fluor 488-labeled Aβ peptides was added and incubated for 4 h at 37 °C in a 5% CO2 atmosphere. Then, cells were washed three times with PBS to arrest uptake, and plasma membrane/cytosol and nuclei were labeled with CellMask™ Orange stain (1:20,000; Thermo Fisher Sci., C10045) and DRAQ5™ (1:500; Thermo Fisher Sci., 62251), respectively, for 5 min. Finally, cells were fixed with 4% PFA and observed under a Leica TCS SPE confocal laser scanning microscope (Leica Microsystems) using a 63x/1.32–0.60 oil PH3 CS objective and a confocal pinhole set at 1 Airy unit. From each well, three non-overlapping images from the top, middle and bottom areas were randomly taken. The ImageJ software [34 (link)] was used to calculate the CTCF by applying the following formula: CTCF = Integrated Density − (Area of selected cell × Mean fluorescence of background readings).
de Dios C., Abadin X., Roca-Agujetas V., Jimenez-Martinez M., Morales A., Trullas R., Mari M, & Colell A. (2023). Inflammasome activation under high cholesterol load triggers a protective microglial phenotype while promoting neuronal pyroptosis. Translational Neurodegeneration, 12, 10.
Publication 2023
Atmosphere Cardiac Arrest Cell Nucleus Cells CTCF protein, human Cytosol Fluorescence GART protein, human Microglia Microscopy, Confocal Peptides Phagocytosis Plasma Membrane SpeA protein, Streptococcus pyogenes Stains
3
Quantifying Microglial Aβ Phagocytosis
Microglial phagocytosis was evaluated by quantifying the degree of internalization of the fluorescent HiLyte Fluor 488-labeled Aβ1-42 peptide (AnaSpec, AS-60479-01). Cells were seeded in 8-well LabTek removable chamber slides. After treatment, 1 μM HiLyte Fluor 488-labeled Aβ peptides was added and incubated for 4 h at 37 °C in a 5% CO2 atmosphere. Then, cells were washed three times with PBS to arrest uptake, and plasma membrane/cytosol and nuclei were labeled with CellMask™ Orange stain (1:20,000; Thermo Fisher Sci., C10045) and DRAQ5™ (1:500; Thermo Fisher Sci., 62251), respectively, for 5 min. Finally, cells were fixed with 4% PFA and observed under a Leica TCS SPE confocal laser scanning microscope (Leica Microsystems) using a 63x/1.32–0.60 oil PH3 CS objective and a confocal pinhole set at 1 Airy unit. From each well, three non-overlapping images from the top, middle and bottom areas were randomly taken. The ImageJ software [34 (link)] was used to calculate the CTCF by applying the following formula: CTCF = Integrated Density − (Area of selected cell × Mean fluorescence of background readings).
de Dios C., Abadin X., Roca-Agujetas V., Jimenez-Martinez M., Morales A., Trullas R., Mari M, & Colell A. (2023). Inflammasome activation under high cholesterol load triggers a protective microglial phenotype while promoting neuronal pyroptosis. Translational Neurodegeneration, 12, 10.
Publication 2023
Atmosphere Cardiac Arrest Cell Nucleus Cells CTCF protein, human Cytosol Fluorescence GART protein, human Microglia Microscopy, Confocal Peptides Phagocytosis Plasma Membrane SpeA protein, Streptococcus pyogenes Stains
4
Macrophage Phagocytosis Assay with IR-61

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Publication 2023
Agar Bacteria Cells Escherichia coli Flow Cytometry Fluorescence Gentamicin Infection Macrophage Microscopy Phagocytosis Staphylococcus aureus Infection Sterility, Reproductive Technique, Dilution Triton X-100
5
Transcriptomic Profiling of Microglia
FACS-sorted microglia cells were flash frozen in dry ice into 300 μl of RNAprotect cell reagent (QIAGEN). Total RNA was extracted using RNeasy mini kit (QIAGEN). RNA samples were sent to the Institute for Genome Sciences at University of Maryland School of Medicine for RNA quality test and RNA-seq. The RNA quality was analyzed using low-input Pico chip on Agilent 2100 Bioanalyzer. Amplification of cDNA from the total RNA samples were performed using Ovation RNA-seq system kit (Tecan), followed by low input RNA-seq library preparation. RNA-seq of all libraries was performed on Illumina HiSeq4000. The data were converted to FastQ format.
Bioinformatics analyses were performed on the High Performance Computing Facility taki CPU cluster at University of Maryland Baltimore County. Data quality was analyzed using FastQC v0.11.8 (available at: http://bioinformatics.babraham.ac.uk/projects/fastqc/). Ribosomal RNA were filtered using SortMeRNA v2.1b (60 (link)). Transcript-level abundance was quantified using the quasi-mapping–based mode in Salmon v1.0.0 (61 (link)), mapping to GRCm38 (mm10) mouse reference genome. Transcript-level abundance were aggregated to gene-level abundance using tximport (62 (link)), and DESeq2 (63 (link)) was used for differential expression analyses. PCA was performed using the mixOmics package (64 (link)). Tximport, DESeq2, and mixOmics packages were run in R v3.6.1 in RStudio v1.2. Pathway enrichment analysis were performed following the protocols described by Reimand et al. (65 (link)). Enriched biological pathways in each group were identified using g:Profiler (66 (link)) based on Gene Ontology biological process database. The biological pathway networks were built on the basis of the g:Profiler results using Cytoscape v3.8.0 (67 (link)). SLEA was performed as previously described at (https://genomemedicine.biomedcentral.com/articles/10.1186/gm327). Briefly, the mean expression of 10,000 random sets of genes the same size as the scored gene set were calculated as used as the null distribution for each sample. The mean expression of the scored gene set was then calculated and transformed to z score based on the null distribution (SLEA score). Gene sets were used from the Gene Ontology Biological Process database for phagocytosis (M16307), regulation of autophagy (M10281), and regulation of ROS biosynthetic process (M15379). DAM/senescence gene set was used from Hu et al. (68 (link)).
Ritzel R.M., Li Y., Jiao Y., Lei Z., Doran S.J., He J., Shahror R.A., Henry R.J., Khan R., Tan C., Liu S., Stoica B.A., Faden A.I., Szeto G., Loane D.J, & Wu J. (2023). Brain injury accelerates the onset of a reversible age-related microglial phenotype associated with inflammatory neurodegeneration. Science Advances, 9(10), eadd1101.
Publication 2023
Anabolism Autophagy Biological Processes Biopharmaceuticals CA-19-9 Antigen Cells DNA, Complementary DNA Chips DNA Library Dry Ice Freezing Gene Expression Genes Genome Microglia Mus Phagocytosis Ribosomal RNA RNA-Seq Salmo salar

Top products related to «Phagocytosis»

1
Cytochalasin d by Merck Group
Sourced in United States, Germany, Macao, United Kingdom, China, Italy, Canada, Sao Tome and Principe
Cytochalasin D is a laboratory reagent that inhibits actin polymerization. It is commonly used in cell biology research to disrupt the cytoskeleton and study its role in cellular processes.
2
Facscalibur by BD
Sourced in United States, Germany, United Kingdom, China, Canada, Japan, Italy, France, Belgium, Switzerland, Singapore, Uruguay, Australia, Spain, Poland, India, Austria, Denmark, Netherlands, Jersey, Finland, Sweden
The FACSCalibur is a flow cytometry system designed for multi-parameter analysis of cells and other particles. It features a blue (488 nm) and a red (635 nm) laser for excitation of fluorescent dyes. The instrument is capable of detecting forward scatter, side scatter, and up to four fluorescent parameters simultaneously.
3
Fetal bovine serum (fbs) by Thermo Fisher Scientific
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Fetal Bovine Serum (FBS) is a cell culture supplement derived from the blood of bovine fetuses. FBS provides a source of proteins, growth factors, and other components that support the growth and maintenance of various cell types in in vitro cell culture applications.
4
Facscanto 2 by BD
Sourced in United States, Germany, United Kingdom, Belgium, China, Australia, France, Japan, Italy, Spain, Switzerland, Canada, Uruguay, Netherlands, Czechia, Jersey, Brazil, Denmark, Singapore, Austria, India, Panama
The FACSCanto II is a flow cytometer instrument designed for multi-parameter analysis of single cells. It features a solid-state diode laser and up to four fluorescence detectors for simultaneous measurement of multiple cellular parameters.
5
Vybrant phagocytosis assay kit by Thermo Fisher Scientific
Sourced in United States
The Vybrant Phagocytosis Assay Kit is a fluorescence-based assay designed to measure the phagocytic activity of cells. The kit provides pre-labeled fluorescent particles that can be engulfed by phagocytic cells, and a fluorescence-based detection method to quantify the phagocytic activity.
6
Phagocytosis assay kit by Cayman Chemical
Sourced in United States
The Phagocytosis Assay Kit is a laboratory tool designed to measure the ability of phagocytic cells, such as macrophages or neutrophils, to engulf and internalize particles or microorganisms. The kit provides the necessary components to facilitate the phagocytosis process and quantify the extent of particle uptake by the target cells.
7
Penicillin streptomycin by Thermo Fisher Scientific
Sourced in United States, Germany, United Kingdom, China, Canada, France, Japan, Australia, Switzerland, Israel, Italy, Belgium, Austria, Spain, Gabon, Ireland, New Zealand, Sweden, Netherlands, Denmark, Brazil, Macao, India, Singapore, Poland, Argentina, Cameroon, Uruguay, Morocco, Panama, Colombia, Holy See (Vatican City State), Hungary, Norway, Portugal, Mexico, Thailand, Palestine, State of, Finland, Moldova, Republic of, Jamaica, Czechia
Penicillin/streptomycin is a commonly used antibiotic solution for cell culture applications. It contains a combination of penicillin and streptomycin, which are broad-spectrum antibiotics that inhibit the growth of both Gram-positive and Gram-negative bacteria.
8
Lipopolysaccharides (lps) by Merck Group
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The LPS laboratory equipment is a high-precision device used for various applications in scientific research and laboratory settings. It is designed to accurately measure and monitor specific parameters essential for various experimental procedures. The core function of the LPS is to provide reliable and consistent data collection, ensuring the integrity of research results. No further details or interpretations can be provided while maintaining an unbiased and factual approach.
9
Facscalibur flow cytometer by BD
Sourced in United States, Germany, United Kingdom, China, Canada, Japan, Belgium, France, Spain, Italy, Australia, Finland, Poland, Switzerland, Cameroon, Uruguay, Denmark, Jersey, Moldova, Republic of, Singapore, India, Brazil
The FACSCalibur flow cytometer is a compact and versatile instrument designed for multiparameter analysis of cells and particles. It employs laser-based technology to rapidly measure and analyze the physical and fluorescent characteristics of cells or other particles as they flow in a fluid stream. The FACSCalibur can detect and quantify a wide range of cellular properties, making it a valuable tool for various applications in biology, immunology, and clinical research.
10
Rpmi 1640 by Thermo Fisher Scientific
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RPMI 1640 is a common cell culture medium used for the in vitro cultivation of a variety of cells, including human and animal cells. It provides a balanced salt solution and a source of essential nutrients and growth factors to support cell growth and proliferation.

More about "Phagocytosis"

Phagocytosis is the crucial cellular process by which specialized immune cells, known as phagocytes, engulf and internalize solid particles, macromolecules, or other matter from their surrounding environment.
This fundamental biological mechanism is essential for immune function, tissue homeostasis, and the clearance of pathogens, cellular debris, and foreign substances.
Key phagocytes involved in this process include macrophages and neutrophils, which play a vital role in engulfing and digesting these materials.
Understanding the intricate mechanisms and regulation of phagocytosis is crucial for unraveling the complexities of immune responses, inflammation, and the pathogenesis of various diseases.
Researchers often utilize tools and techniques like the FACSCalibur flow cytometer, Vybrant Phagocytosis Assay Kit, and Phagocytosis Assay Kit to study phagocytic activity.
Culturing cells in RPMI 1640 medium supplemented with FBS and Penicillin/streptomycin can also support these investigations.
Additionally, substances like Cytochalasin D and LPS can be employed to modulate phagocytosis and elucidate its underlying pathways.
Advances in research tools and techniques, such as PubCompare.ai, can greatly enhance phagocytosis studies by helping researchers identify the most reproducible and accurate protocols from the available literature, preprints, and patents.
This AI-driven approach boosts productivity and accelerates scientific discoveries in this critical area of cell biology, empowering researchers to take their phagocytosis research to new heights.