The intracellular calcium assay is a method for the determination of intracellular calcium. It has a long history of use in many fields of biology and medicine, where it is a valuable method for testing the effects of drugs, chemicals and other biological agents on cellular calcium levels. It is also used extensively to test the efficacy of medical drugs in modulating the intracellular calcium level. The most widely used technique for this is the alkaline phosphate precipitation method, whereby alkaline phosphate is mixed with a cell culture medium and then mixed with the sample of interest. The final mixture is then subjected to an ultra-violet light, which will eliminate any dissolved molecules that are present in the sample.
Benefits Associated With The Intracellular Calcium Assay
There are several advantages associated with the intracellular calcium assay: the test is sensitive to small changes in the concentration of intracellular calcium; normally no change in the concentration of intracellular calcium is detected when cells are frozen; normally the tested cells are plucked and frozen immediately after collection; the procedure is simple, requiring no special equipment or skill; and reliable results are obtained. It is possible to prepare and use the same test in different types of cells, including both the living and the dead. In this manner, the test is applicable to a wide range of different applications.
The most commonly used method for this type of assay is the standard glass microscope/liquid stage hybrid method. Briefly, glass slide suspension cells are placed on slides and lids are used to prevent them from being scratched. A solution of dye is added on top of the suspension cells, which are allowed to become immersed in the dye solution overnight. On the day following the overnight treatment, the dye concentration in the suspension cells is detected and the concentration of each particular dye in the intracellular calcium culture medium is determined.
Usage In Medical Research
The next type of intracellular calcium assays employed in medical research is the automated cell washing system. In this method, cells are washed in chlorinated water; a high throughput filter, containing multiple carbon blocks, is then fitted over the wells. A light chemical colorant is added, followed by high-throughput immunofluorescence methods to identify the individual cells. The final step of this method is to pass the cells through a potassium permeation network and observe if the light chemical colorant interacts with the antigens that elicit a personal immunity response. The major advantage of this method is the speed of completing the assay; the sample is passed through the filter, incubated for a few hours in the laboratory, and the final measurement performed on a day in the lab when the sample is returned at the temperature of the incubation vessel.
The third intracellular calcium assays commonly used in medical research are the local library system and the polymyalgia rheumatica library system. The local library system combines two different systems: local, which is a highly selective and durable liquid suspension; and mar ants, which contain various sources of local and have been studied extensively using the high throughput screening methods. The combination allows researchers to combine two different assays in one well-designed system. This method also allows researchers to test the effects of multiple substances on a single cell without requiring two separate solutions. The local library system uses local tubes with vials; multiple vials can be used simultaneously to generate a higher level of sensitivity.
The polymyalgia rheumatica assay has established efficacy and reproducibility compared to the other two established assays using freshly detached suspension cells. To make this method even more suitable, researchers coating the plates with different substances that can induce immunity (for instance, purified antibodies or purified phytoestrogens). This method may be a potential treatment for the disease, although further studies must be conducted to determine its effectiveness and safety profile.
