Hi Narender,

Typically you load the cells with a cell-permeable fluorophore like BAPTA. It binds to intracellular Ca and this changes its emission spectrum. You can use any read-out: fluorescent ELISA or flow cytometry, whichever is available. There is a variety of Ca--binding fluorophores on the market, and the protocols are easy.

Regards,

Yelena 

Physiol Rev. 1999 Oct;79(4):1089-125.

Measurement of intracellular calcium.

Takahashi A1, Camacho P, Lechleiter JD, Herman B.

Author information

Abstract

To a certain extent, all cellular, physiological, and pathological phenomena that occur in cells are accompanied by ionic changes. The development of techniques allowing the measurement of such ion activities has contributed substantially to our understanding of normal and abnormal cellular function. Digital video microscopy, confocal laser scanning microscopy, and more recently multiphoton microscopy have allowed the precise spatial analysis of intracellular ion activity at the subcellular level in addition to measurement of its concentration. It is well known that Ca2+ regulates numerous physiological cellular phenomena as a second messenger as well as triggering pathological events such as cell injury and death. A number of methods have been developed to measure intracellular Ca2+. In this review, we summarize the advantages and pitfalls of a variety of Ca2+ indicators used in both optical and nonoptical techniques employed for measuring intracellular Ca2+ concentration.

A blood calcium test is ordered to screen for, diagnose, and monitor a range of conditions relating to the bones, heart, nerves, kidneys, and teeth. The test may also be ordered if a person has symptoms of a parathyroid disorder, malabsorption, or an overactive thyroid.

A total calcium level is often measured as part of a routine health screening. It is included in the comprehensive metabolic panel (CMP) and the basic metabolic panel (BMP), groups of tests that are performed together to diagnose or monitor a variety of conditions.

When an abnormal total calcium result is obtained, it is viewed as an indicator of an underlying problem. To help diagnose the underlying problem, additional tests are often done to measure ionized calcium, urine calcium, phosphorous, magnesium, vitamin D, parathyroid hormone (PTH) and PTH-related peptide (PTHrP). PTH and vitamin D are responsible for maintaining calcium concentrations in the blood within a narrow range of values.

If the calcium is abnormal, measuring calcium and PTH together can help determine whether the parathyroid glands are functioning normally. Measuring urine calcium can help determine whether the kidneys are excreting the proper amount of calcium, and testing for vitamin D, phosphorus, and/or magnesium can help determine whether other deficiencies or excesses exist. Frequently, the balance among these different substances (and the changes in them) is just as important as the concentrations.

Calcium can be used as a diagnostic test if a person has symptoms that suggest:

Kidney stones

Bone disease

Neurologic disorders

The total calcium test is the test most frequently ordered to evaluate calcium status. In most cases, it is a good reflection of the amount of free calcium present in the blood since the balance between free and bound is usually stable and predictable. However, in some people, the balance between bound and free calcium is disturbed and total calcium is not a good reflection of calcium status. In these circumstances, the measurement of ionized calcium may be necessary. Some conditions where ionized calcium should be the test of choice include: critically ill patients, those who are receiving blood transfusions or intravenous fluids, patients undergoing major surgery, and people with blood protein abnormalities like low albumin.

Large fluctuations in ionized calcium can cause the heart to slow down or to beat too rapidly, can cause muscles to go into spasm (tetany), and can cause confusion or even coma. In those who are critically ill, it can be extremely important to monitor the ionized calcium level in order to be able to treat and prevent serious complications.