Densitometer Operation and Principle

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Questions and Answers

What is the primary focus of the interpretation of serum protein electrophoresis?

The subsets of proteins and their relative quantity (correct)

The pH of the buffer solution

The absolute quantity of serum proteins

The type of electrodes used

Which protein component of serum has the largest peak?

Albumin (correct)

Alpha-1 globulin

Beta-2 globulin

Globulin

What is the purpose of using a specific buffer with pH 8.5 in serum protein electrophoresis?

To alter the pH of the serum

To increase the resolution of the electrophoresis

To separate the β fraction into two zones (correct)

To separate albumin from globulins

Which of the following is closest to the negative electrode in a typical serum protein electrophoresis?

Gamma globulin Signup and view all the answers

What is the result of changes in the relative concentration of fractions in serum protein electrophoresis?

Easy recognition of pathological disorders Signup and view all the answers

Which of the following is NOT a subset of globulins in serum protein electrophoresis?

Albumin Signup and view all the answers

What is the main characteristic of the nephrotic pattern in serum electrophoretic patterns?

A decrease in albumin and γ-globulin Signup and view all the answers

What is the effect of the damaged nephron on the serum proteins in nephrotic syndrome?

A loss of albumin and other low molecular weight proteins Signup and view all the answers

Which of the following proteins is increased in the nephrotic pattern?

α2-globulin Signup and view all the answers

What is the role of the densitometer in serum electrophoretic patterns?

To measure light transmittance through a solid sample Signup and view all the answers

How is quantitation performed in serum electrophoretic patterns?

By determining the area of each band as a percent of the total area Signup and view all the answers

What is the function of microprocessors in serum electrophoretic patterns?

To automatically integrate and compute the area under each peak Signup and view all the answers

What is the significance of Arne Tiselius' research?

He researched on electrophoresis and adsorption analysis, especially concerning the complex nature of serum proteins Signup and view all the answers

What is the principle behind electrophoresis?

Separation of molecules based on their charge, shape, and size Signup and view all the answers

What is the application of electrophoresis in biochemistry and molecular biology?

To separate and identify macromolecules based on size, shape, or isoelectric point Signup and view all the answers

What is the purpose of applying an electric field in electrophoresis?

To migrate charged molecules through a gel matrix Signup and view all the answers

What is the advantage of electrophoresis over other separation techniques?

It is highly versatile and can separate molecules based on size, shape, or isoelectric point Signup and view all the answers

What is the significance of serum protein electrophoresis?

It is used to analyze serum proteins, which can aid in diagnosis of various diseases Signup and view all the answers

Study Notes

Serum Protein Electrophoresis

Serum protein electrophoresis results depend on the fractions of two major types of protein: albumin and globulins.
Albumin is the major protein component of serum, produced by the liver under normal physiologic conditions.
Globulins comprise a smaller fraction of the total serum protein content.

Typical Normal Pattern

Albumin forms the largest peak, closest to the positive electrode.
The next components (globulins) are labeled alpha1, alpha2, beta, and gamma, with the gamma peak being closest to the negative electrode.
The use of a specific buffer (pH 8.5) allows separation of the β fraction into two zones: β1 and β2.

Densitometry

A densitometer is a special type of spectrophotometer used to measure light transmittance through a solid sample (electrophoretic strip).
The electrophoretic strip is moved through a measuring optical system, and absorbance of each band is measured.
The area of each fraction is displayed on a strip chart recorder, with each peak representing an individual band on the electrophoretic strip.
Quantitation is performed by determining the area of each band as a percent of the total area for that scan.

Abnormal Patterns

Nephrotic syndrome pattern results from loss of serum proteins through damaged nephrons.
Characterized by a decrease in albumin, α1, and gamma globulins, and an increase in certain large molecular weight proteins (macroglobulin, IgM, lipoproteins).
α2 globulin band is prominently increased.

Theoretical Principles

Electrophoresis is a versatile tool used to separate and identify macromolecules (DNA, RNA, proteins) based on size, shape, or isoelectric point.
Separation of molecules by electrophoresis is based on the fact that charged molecules migrate through a gel matrix upon application of an electric field.

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Description

Learn about the working principle and operation of a densitometer, a special type of spectrophotometer used to measure light transmittance through solid samples like electrophoretic strips. Understand how it measures absorbance and displays results on a strip chart recorder.