4. Here is an immunoassay to measure the explosive trinitrotoluene (TNT) in soil. An organic solvent is used to extract TNT from the soil and then the solvent is analyzed using a flow cytometer. A flow cytometer is a device used to count small particles that flow through a narrow tube past a detector. The particles are irradiated with a green laser and the fluorescence from each particle is measured as it flows past the detector. Step 1. Antibodies that bind TNT are chemically attached to 5-μm latex beads. Step 2. The beads are incubated with fluorescent derivative of TNT to saturate the antibodies and excess TNT is washed away. The beads are suspended in aqueous detergent. Step 3. A volume of 5 μL of the suspension is added to 100 μL of sample or standard containing TNT. TNT in the sample displaces some of the fluorescent TNT. Step 4. An aliquot is injected into the flow cytometer, which measured the fluorescence of the individual beads as they pass by the detector. a) The figure shows the median fluorescence intensity ± the standard deviation of the signal in the range 0.01 -10,000 ppb TNT. Explain why the fluorescence intensity decreases with increasing concentration of TNT. The fluoresnce intensity decreases with increasing concentration TNT as less antibodies are around to detect?
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4. Here is an immunoassay to measure the explosive trinitrotoluene (TNT) in soil. An organic solvent is used to extract TNT from the soil and then the solvent is analyzed using a flow cytometer. A flow cytometer is a device used to count small particles that flow through a narrow tube past a detector. The particles are irradiated with a green laser and the fluorescence from each particle is measured as it flows past the detector.
Step 1. Antibodies that bind TNT are chemically attached to 5-μm latex beads.
Step 2. The beads are incubated with fluorescent derivative of TNT to saturate the antibodies and excess TNT is washed away. The beads are suspended in aqueous detergent.
Step 3. A volume of 5 μL of the suspension is added to 100 μL of sample or standard containing TNT. TNT in the sample displaces some of the fluorescent TNT.
Step 4. An aliquot is injected into the flow cytometer, which measured the fluorescence of the individual beads as they pass by the detector.
a) The figure shows the median fluorescence intensity ± the standard deviation of the signal in the range 0.01 -10,000 ppb TNT. Explain why the fluorescence intensity decreases with increasing concentration of TNT.
The fluoresnce intensity decreases with increasing concentration TNT as less antibodies are around to detect?
Immunoassay is a biochemical technique that can detect the concentration of target molecules by antibody binding. This technique is highly sensitive and can detect a minimal concentration of the target molecules.
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Explain why the fluorescence signal decreases as the concentration of TNT in the sample increases.
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