Profiling single-cell level phagocytic activity distribution with blood lactate levels

Abstract

Investigating the relationship between neutrophil phagocytic activity and blood lactate levels by employing single-cell data.

Bioelectronic Sensor with Magnetic Modulation to Quantify Phagocytic Activity of Blood Cells Employing Machine Learning

“…These particles are roughly the size of an average bacterial cell (∼0.5 μm mean diameter) and coated with the same ligand (IgG) that would be present on a bacterial cell as the immune system gets activated and produces antibodies in response to bacterial infection. Opsonized microparticles were also used in the past by many researchers to quantify the phagocytic activity of blood cells instead of using specific pathogens. ,− In the antibody-mediated phagocytosis process, the receptor molecules (i.e., CD64) on the neutrophils will bind to the particles in the sample and begin the phagocytosis process including internalization and killing. However, since the particles were made of a non-organic ferrous material, they cannot be digested by the cells and will instead remain indefinitely trapped within a vesicle.…”

“…Neutrophils, the most abundant phagocytes present in the human blood, were isolated from each blood sample. An isolation protocol developed to achieve this is based on our earlier studies . First, the whole blood samples were diluted 50% with 1× phosphate-buffered saline (PBS).…”

“…An isolation protocol developed to achieve this is based on our earlier studies. 23 First, the whole blood samples were diluted 50% with 1× phosphate-buffered saline (PBS). A total of 0.8 mL of these diluted whole blood samples was pipetted on top of 0.6 mL of Ficoll Plaque density gradient, as shown on the left side of Figure S6, and the samples were centrifuged at 400g (∼3920 m/s 2 ) for 25 min.…”

Antibody-functionalized aluminum oxide-coated particles targeting neutrophil receptors in a multifrequency microfluidic impedance cytometer

“…The previous step was repeated until a neutrophil pellet formed, which was resuspended in RPMI 1640 with 50 μL of cells to 5 mL of media. 49…”

“…The previous step was repeated until a neutrophil pellet formed, which was resuspended in RPMI 1640 with 50 μL of cells to 5 mL of media. 49 Once prepared, a 200 μL solution of either 10 nm Al 2 O 3 coated-Janus microparticles with functionalized anti-CD11b antibody (10 nm MOJPs/anti-CD11b) or 30 nm Al 2 O 3 coated-Janus microparticles with functionalized anti-CD66b antibody (30 nm MOJPs/anti-CD66b) at 5.92 × 10 7 particles per mL each were mixed with 1 mL of cells diluted in 1× PBS followed by 1 hour incubation. For samples without MOJPs added (cells alone), the isolated neutrophils were diluted in 1× PBS and incubated for 1 hour.…”