This set will teach you about the basic instrumentation covered on the MLT/MLS ASCP registry exam. We will go over multiple different analyzers and the methodologies for each test.
Terms in this set (30)
Explain the principle of a normal sandwich ELISA
The primary (or capture) antibody is bound to a solid phase. Then the sample is introduced and the antigen binds to the primary antibody. Then the cuvette is washed out and only the bound antibody and antigen are left. Then a labeled secondary (or test) antibody is added which binds to the antigen then the label is detected.
an excess of antibodies which results in a lack of agglutination/precipitation despite the presence of an antigen. The abundance of antibodies causes each antigen to bind a different antibody, which impairs the ability for the antibodies to crosslink and cause agglutination/precipitation. One antibody must bind two different antigens to crosslink.
an excess of antigen which results in a lack of agglutination/precipitation despite the presence of an antigen. The abundance of antigen floods the binding sites of the antibodies and impairs the ability to crosslink and cause agglutination/precipitation.
How do interfering antibodies cause false negatives?
They bind to the antigen instead of the test antibody
How to interfering antibodies cause false positives?
They bind to something that you're not interested in which is attached to the solid phase. Then if the testing antibody is a labeled anti-IgG, then the interfering antibody will be detected and produce a false signal
What is the capture antibody attached to?
What is a good analogy fo adsorption?
Boats with metal sides floating in the water with magnets along the sides. The speed of the current and attraction to the magnets determine if the boat will float downstream or to the side
Define the Rf in Thin Layer Chromatography
The distance that the solute traveled divided by the distance that the solvent front traveled.
List the parts of an HPLC in order
Explain the general principles of Mass spectrometry
Ionization, Acceleration, Deflection, Detection
Describe the costs of automation in clinical chemistry
Potential reduction in workforce
Describe the benefits of automation in clinical chemistry
What is the principle behind spectrophotometry?
Production of light by a light source, diffraction of light, passed through an adjustible aperture, through the sample, and detected at specific wavelengths.
Detection of light is much more precise than the human eye.
Describe how Beer's law relates to clinical chemistry
The relationship between the absorption of light by a solution is proportional to the concentration of the target in that solution
Transmittance (%T) = Transmitted light (T)/ Incident light (To)
Absorption (A) = abc
We have to re-learn our ABCs
a = molar absorptivity constant
b = light path through the solution
c = concentration of solute
What is the equation from Absorbance to %T?
A = 2-log%T
Hint: it's a virtual guarantee to get a question on this on the ASCP registry exam
Why do we want to keep absorbance values below 1?
Because we lose sensitivity in the test when absorbance values increase above 1.
What is the Absorbance if the transmittance of a specimen blank is 0.981 and the sample is 0.647?
%T = Transmitted light / Incident light
66% = 0.647 / 0.981 x 100
A = 2 - log %T
2.18 = 2 - (-0.181)
Measures the light blocked or reflected by particles in suspension. Uses the same principle as a standard spectrohpotometer The image on the right is more turbid than the image on the left
Incident light is NOT measured. Reflected light is measured at 90˚ also called side scatter or at 45˚ called forward scatter
Measures current between two electrodes. Used in the clarke pO2 electrode
Measures the potential (duh) between two electrodes. Used in the pCO2 (Severinghaus) and pH electrodes
Quantifies the amount of substance that can be oxidized. Results in complete analyte consumption
Quantifies the amount of substance that generates a current. Results in NO analyte consumption
What stands out in a pH meter?
contains a selectively permeable glass membrane and a Ag/AgCl reference electrode
What semipermeable membrane allows for K+ to cross into the measuring chamber?
Valinomycin is the key word for this and is used in K+ ion-selective electrodes
Explain the principle behind electrophoresis
Think of it as dragging a bunch of rocks across a big field.
It is easier to move the lighter ones farther
If they are the right shape for rolling, they will move farther as well.
The Larger and Lower charged molecules don't move very far from the application site, but the highly charged and smaller molecules shoot out like a rocket.
Name the four colligative properties
Colligative properties are dependent upon what?
The concentration of the solute, NOT the identity of the solute
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