In Vitro Cell Viability by the Lactate Dehydrogenase Assay LDH 2

In this experiment we will determine the activity of the cytoplasmic enzyme lactate dehydrogenase (LDH) released by damaged cells after exposure to the geometric concentration of different nano particles.

Learning Objectives (ILOs)
1- Successfully handle the required instruments and consumables needed in the experiment.
2- Check the confluence and count cell under the microscope.
3- Dilute the cells to a specific count suitable for seeding in the 96-well plate .
4- Calculate the concentration of tested chemicals and prepare the calculated doses in the cell culture medium.
5- Aspirate the old medium and add the new medium containing the tested chemicals in the appropriate wells.
6- Add the LDH assay substrate solution to cells and read the results using the microplate reader after incubation of cells.
7- Prepare desired concentrations of LDH standard solution in cell culture medium, and draw the standard curve.
8- Read the results of LDH and calculate the cytotoxicity percent for cells exposed to different doess of tested chemicals.
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