You perform an ELISA to test hospital soap samples for contamination with Pseudomonas aeruginosa. Other than the different molecule of interest, this ELISA works exactly like the one you saw in the video. You run the soap sample, a negative control, and a positive control, all in duplicate. How would you interpret the following results?   Laboratory Notebook: ELISA – Pseudomonas aeruginosa Well Contents Observation at 15 min Result 1 Negative control     2 Soap sample     3 Positive control     4 Negative control     5 Soap sample     6 Positive control     7 Empty well     8 Empty well           The soap is POSITIVE for the Pseudomonas aeruginosa antigen. You cannot interpret the soap data, because the NEGATIVE control didn’t work. The soap is NEGATIVE for the Pseudomonas aeruginosa antigen. You cannot interpret the soap data, because the POSITIVE control didn’t work.

Basic Clinical Laboratory Techniques 6E
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ISBN:9781133893943
Author:ESTRIDGE
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Chapter4: Basic Immunology And Immunohematology
Section4.3: Tests For Rheumatoid Factors
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You perform an ELISA to test hospital soap samples for contamination with Pseudomonas aeruginosa. Other than the different molecule of interest, this ELISA works exactly like the one you saw in the video. You run the soap sample, a negative control, and a positive control, all in duplicate. How would you interpret the following results?

 

Laboratory Notebook: ELISA – Pseudomonas aeruginosa
Well Contents Observation at 15 min Result
1 Negative control    
2 Soap sample    
3 Positive control    
4 Negative control    
5 Soap sample    
6 Positive control    
7 Empty well    
8 Empty well    

 

 

 

The soap is POSITIVE for the Pseudomonas aeruginosa antigen.
You cannot interpret the soap data, because the NEGATIVE control didn’t work.
The soap is NEGATIVE for the Pseudomonas aeruginosa antigen.
You cannot interpret the soap data, because the POSITIVE control didn’t work.
The image showcases a series of cuvettes commonly used in spectrophotometric analysis, held in a testing rack. Each of the eight cuvettes is filled with a solution of varying concentrations, indicated by the different intensities of the blue-green color in the fluid inside them. The cuvettes are labeled with the numbers 1 through 8 written in blue marker on their upper parts.

### Detailed Explanation:

1. **Cuvettes and Their Use**: 
   - Cuvettes are small, rectangular containers used in spectrophotometry to hold samples. They are designed to fit into a spectrophotometer, an instrument that measures how much light a sample absorbs.

2. **Numbering and Concentration**:
   - Each cuvette in the image is clearly numbered (1 to 8) with blue ink. The numbers appear to correlate with the concentrations of the solutions they contain. Typically, such numbering would be part of a method to systematically analyze different concentration levels of a substance.

3. **Color intensities**:
   - The color variation from light to dark blue-green indicates different concentration levels of a solute in the solvent. Cuvettes 1 and 2 display lighter shades, suggesting lower concentrations, whereas cuvettes 3 through 6 show progressively darker shades, indicative of higher concentrations. Cuvettes 7 and 8 appear to be empty, likely serving as controls or blanks.

4. **Spectrophotometric Analysis**:
   - During spectrophotometric analysis, light is passed through these solutions, and the amount of light absorbed by the solution is measured. The absorbance data can then be used to determine the concentration of the solute in each cuvette, following Beer’s Law. This principle states that the absorbance is directly proportional to the concentration of the absorbing species in the solution.

### Educational Context:

Understanding how to prepare and analyze samples using spectrophotometry is crucial in various scientific fields, including biochemistry, molecular biology, and environmental science. The setup shown in the image can be part of a laboratory exercise for students to learn how to correlate the intensity of color (absorbance) with the concentration of a solution.

### Conclusion:

By conducting experiments with cuvettes containing solutions of known concentrations, students can create a standard curve, which is essential for determining the concentration of unknown samples in future experiments. The image exhibits a practical example of setting up such an experiment, emphasizing the importance of careful
Transcribed Image Text:The image showcases a series of cuvettes commonly used in spectrophotometric analysis, held in a testing rack. Each of the eight cuvettes is filled with a solution of varying concentrations, indicated by the different intensities of the blue-green color in the fluid inside them. The cuvettes are labeled with the numbers 1 through 8 written in blue marker on their upper parts. ### Detailed Explanation: 1. **Cuvettes and Their Use**: - Cuvettes are small, rectangular containers used in spectrophotometry to hold samples. They are designed to fit into a spectrophotometer, an instrument that measures how much light a sample absorbs. 2. **Numbering and Concentration**: - Each cuvette in the image is clearly numbered (1 to 8) with blue ink. The numbers appear to correlate with the concentrations of the solutions they contain. Typically, such numbering would be part of a method to systematically analyze different concentration levels of a substance. 3. **Color intensities**: - The color variation from light to dark blue-green indicates different concentration levels of a solute in the solvent. Cuvettes 1 and 2 display lighter shades, suggesting lower concentrations, whereas cuvettes 3 through 6 show progressively darker shades, indicative of higher concentrations. Cuvettes 7 and 8 appear to be empty, likely serving as controls or blanks. 4. **Spectrophotometric Analysis**: - During spectrophotometric analysis, light is passed through these solutions, and the amount of light absorbed by the solution is measured. The absorbance data can then be used to determine the concentration of the solute in each cuvette, following Beer’s Law. This principle states that the absorbance is directly proportional to the concentration of the absorbing species in the solution. ### Educational Context: Understanding how to prepare and analyze samples using spectrophotometry is crucial in various scientific fields, including biochemistry, molecular biology, and environmental science. The setup shown in the image can be part of a laboratory exercise for students to learn how to correlate the intensity of color (absorbance) with the concentration of a solution. ### Conclusion: By conducting experiments with cuvettes containing solutions of known concentrations, students can create a standard curve, which is essential for determining the concentration of unknown samples in future experiments. The image exhibits a practical example of setting up such an experiment, emphasizing the importance of careful
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