Applies ‘local neighborhood processing’ in order to compensate for uneven background on fluoro plates. This way the spot volume is quantified, independent of the local background.

Small volume, higher throughput.

Blue, red, green or silver substrat Elispot.

Detection antibody, enzyme-conjugate and precipitating substrate.

"The enzyme-linked immune absorbent spot (ELISpot) is a type of assay that focuses on quantitatively measuring the frequency of cytokine secretion for a single cell."

[https://en.wikipedia.org/wiki/ELISpot]

Blue and red Elispot substrate (mixed color violet)

Dual cytokine secretion.

"Using DNA recombinant technology, scientists combine the RFP gene to a another gene that produces a protein that they want to study, and then they insert the complex into a cell." [https://embryo.asu.edu/pages/green-fluorescent-protein but with RFP instead of GFP]

"Typically, type 1 alveolar cells comprise the major gas exchange surface of the alveolus and are integral to the maintenance of the permeability barrier function of the alveolar membrane. Type 2 pneumocytesare the progenitors of type 1 cells and are responsible for surfactant production and homeostasis." [https://www.sciencedirect.com/topics/medicine-and-dentistry/alveolar-type-i-cells]

The alveolar epitheliumis a major target in toxic exposures of the lung because of its structural delicacy and proximity to inhaled toxicants. Type II epithelial cellsare important in maintaining the integrity of alveolar epitheliumand normal lung function.

"Cytomegalovirus (CMV) (from the Greek cyto-, "cell," and megalo-, "large") is a genus of viruses in the order Herpesvirales." [https://en.wikipedia.org/wiki/Cytomegalovirus]

MCMV mutants, tdTomato, Cherry, eGFP fibroplasts

"The glycoprotein gO (UL74) of human cytomegalovirus (HCMV) forms a complex with gH/gL. Virus mutants with a deletion of gO show a defect in secondary envelopment with the consequence that virus spread is restricted to a cell-associated pathway." [https://www.ncbi.nlm.nih.gov/pubmed/20181688]

"Human embryonic kidney 293 cells, also often referred to as HEK 293, HEK-293, 293 cells, or less precisely as HEK cells, are a specific cell line originally derived from human embryonic kidney cellsgrown in tissue culture." [https://en.wikipedia.org/wiki/HEK_293_cells]

"Natural killer cells, or NK cells, are a type of cytotoxic lymphocyte critical to the innate immune system. Typically, immune cells detect major histocompatibility complex (MHC) presented on infected cell surfaces, triggering cytokine release, causing lysis or apoptosis." [https://en.wikipedia.org/wiki/Natural_killer_cell]

"Natural killer cells, or NK cells, are a type of cytotoxic lymphocyte critical to the innate immune system. Typically, immune cells detect major histocompatibility complex (MHC) presented on infected cell surfaces, triggering cytokine release, causing lysis or apoptosis." [https://en.wikipedia.org/wiki/Natural_killer_cell]

"The spleen is an organ found in virtually all vertebrates. Similar in structure to a large lymph node, it acts primarily as a blood filter. A study published in 2009 using mice found that the red pulp of the spleen forms a reservoir that contains over half of the body's monocytes." [https://en.wikipedia.org/wiki/Spleen]

"The focus forming assay (FFA) is a variation of the plaque assay, but instead of relying on cell lysis in order to detect plaque formation, the FFA employs immunostaining techniques using fluorescently labeled antibodies specific for a viral antigen to detect infected host cells and infectious virus particles before an actual plaque is formed." [https://en.wikipedia.org/wiki/Virus_quantification]

"The focus forming assay (FFA) is a variation of the plaque assay, but instead of relying on cell lysis in order to detect plaque formation, the FFA employs immunostaining techniques using fluorescently labeled antibodies specific for a viral antigen to detect infected host cells and infectious virus particles before an actual plaque is formed." [https://en.wikipedia.org/wiki/Virus_quantification]

"Using DNA recombinant technology, scientists combine the GFP gene to a another gene that produces a protein that they want to study, and then they insert the complex into a cell." [https://embryo.asu.edu/pages/green-fluorescent-protein]

"The TCID50 (Median Tissue Culture Infectious Dose) is one of the methods used when verifying viral titer. TCID50 signifies the concentration at which 50% of the cells are infected when a test tube or well plate upon which cells have been cultured is inoculated with a diluted solution of viral fluid." [https://www.zeomic.co.jp/en/glossary/virus/71]

Cells were infected and covered with an overlay. Surviving cells stained with CV. Cells killed by the virus.

Cells were infected and covered with an overlay. Surviving cells stained with CV. Cells killed by the virus.

"The plaque reduction neutralization test is used to quantify the titer of neutralizing antibody for a virus. The serum sample or solution of antibody to be tested is diluted and mixed with a viral suspension." [https://en.wikipedia.org/wiki/Plaque_reduction_neutralization_test]

"An organoid is a miniaturized and simplified version of an organ produced in vitro in three dimensions that shows realistic micro-anatomy. The technique for growing organoids has rapidly improved since the early 2010s." [https://en.wikipedia.org/wiki/Organoid]

A segment of DNA introduced to some other organism. See [https://en.wikipedia.org/wiki/Transgene]