Content1. What is electrophoresis apparatus?
1.1 Working principle of electrophoresis apparatus
1.2 History of electrophoresis apparatus2. Type of electrophoresis apparatus
2.1 Gel electrophoresis apparatus3. Use of electrophoresis apparatus
3.1 Notes on the use of electrophoresis apparatus
3.2 Common troubleshooting of electrophoresis apparatus4. How to buy electrophoresis apparatus?
What is electrophoresis apparatus?
is an instrument that realizes electrophoretic analysis
. It generally consists of a power supply, electrophoresis tank, detection unit, etc. The so-called electrophoresis refers to the movement of charged particles in the electric field. Different substances move at different speeds in the electric field because of their different charges and molecular weights, according to which different substances can be analyzed qualitatively or quantitatively, a certain mixture can be analyzed in components or individual components can be extracted and prepared.The electrophoresis apparatus
can be used for qualitative or quantitative analysis
of different substances, component analysis of a certain mixture, or extraction and preparation of individual components. It is often used in laboratory tests in clinical medicine or scientific experimental studies.
Working principle of electrophoresis apparatus
The particles of substances that can adsorb charged masses or have dissociable groups in solution, such as proteins and amino acids, are bound to be attracted by electrodes with opposite electrical properties and move in a DC electric field under certain pH conditions. In addition to the charged state and electric field strength, the moving speed of particles of different substances in the electric field is also related to the particles' size, shape, and medium viscosity. According to this feature, the application of electrophoresis can be a qualitative or quantitative analysis of different substances or a certain mixture for component analysis or individual component extraction preparation, electrophoresis instrument is designed and manufactured based on the above principle.
There are many factors affecting electrophoresis, mainly the amount of charge of the separated substance, the strength of the electric field, the pH and ionic strength of the buffer, and the chemical inertness of the supporting medium.
History of electrophoresis apparatus
Since the first commercialized boundary-shifting electrophoresis system was developed by Swedish physical chemist Professor Tiselius in 1946, the development of electrophoresis analyzers has been extremely rapid. Since the 1970s, more and more automated electrophoresis analyzers have been introduced into clinical laboratories and have been playing an increasingly important role in the clinical diagnosis and treatment of various diseases.
A. Early stage (before ~1970)
The two main types of supporting media are cellulose acetate film (referred to as acetate film) and agarose gel, and electrophoresis apparatuses in this period were mostly used for the analysis of proteins such as serum proteins, lipoproteins, hemoglobin, and other items.
B. Middle stage (1970~1990)
In this period, electrophoresis apparatuses were mainly visible light/fluorescent dual system automatic electrophoresis scanners. The introduction of fluorescent reagents and fluorescent scanners greatly facilitated the determination of lactate dehydrogenase (LD) and creatine kinase (CK) isoenzymes in clinical practice.
C. Development stage (1990~present)
In this period, electrophoresis apparatuses were based on automation.
Type of electrophoresis apparatus
Gel electrophoresis apparatus
The electrophoresis is divided into free electrophoresis and zonal electrophoresis according to whether a support medium is used in electrophoresis.
Free electrophoresis does not use a support medium, and electrophoresis is performed in solution. This type of electrophoresis is further divided into two categories: non-free interface electrophoresis and free interface electrophoresis. Non-free interface electrophoresis means that all charged particles (such as various cells) suspended in the solution move after electrification and no interface appears, such as microelectrophoresis. In free interface electrophoresis, the separated substances are concentrated in a certain layer and form their interface for qualitative or quantitative analysis. Free interface electrophoresis requires expensive and sophisticated current instruments and is used only in a few special electrophoresis applications such as isoelectric focusing electrophoresis and isotropic electrophoresis.
All zonal electrophoresis use support media, which are divided into filter paper electrophoresis, acetate membrane electrophoresis, thin layer electrophoresis, and gel electrophoresis depending on the support media. In addition, the different forms of devices of support medium can be divided into horizontal plate electrophoresis, vertical plate electrophoresis, vertical disk electrophoresis, capillary electrophoresis, bridge electrophoresis, and continuous flow electrophoresis, etc.
Several common types of electrophoresis apparatus are as follows.
A. Fully automatic fluorescence/visible light dual system electrophoresis instrument
Automatic fluorescence/visible light dual system electrophoresis instrument has a fluorescence/visible light dual system, using fluorescent reagent items, such as CK, and LD isoenzymes. It has the advantages of high sensitivity, and high accuracy and adopts high pressure and low-temperature system, which is very fast.
B. Automatic vinyl membrane
The fully automated vinyl film electrophoresis instrument is a visible light single system. It uses a vinyl membrane electrophoresis sheet and has the advantage of higher automation. This instrument is mostly used for clinical routine serum protein electrophoresis analysis.
C. Automatic agarose electrophoresis instrument
The automatic agarose electrophoresis instrument is a visible light single system. It uses agarose gel electrophoresis film, which has the advantage of high sensitivity and can be used for low-concentration protein tests, such as urine protein and cerebrospinal fluid protein. The separation of isoenzymes is also quite good, and it can do more items with higher sensitivity.
D. Automatic electrophoresis analysis system
The electrophoresis instrument concentrates the advantages of the above instruments, automatically spotting, electrophoresis, coloring (or staining, decolorization), and drying. It can be used for various electrophoresis sheets, including agar sheets, acetate sheets, polyacrylamide, etc. It adopts the dual system of visible light and fluorescence color presentation, which is a more ideal electrophoresis instrument.
Use of electrophoresis apparatus
a. Connect the two electrodes of the electrophoresis tank to the DC output of the electrophoresis instrument with wires, pay attention not to reverse the polarity.
b. Adjust the power switch of the electrophoresis instrument to the off position, turn the voltage knob to the minimum, and choose the voltage and current range according to the working need.
c. Turn on the power, slowly rotate the voltage adjusting knob until the required voltage is reached, set the electrophoresis termination time, and then start electrophoresis.
d. After the work is finished, turn the knobs and switches to zero or off, and dial out the electrophoresis plug.
Notes on the use of electrophoresis apparatus
a. After the electrophoresis apparatus is energized into working condition, it is forbidden for the human body to touch the electrode, electrophoresis object, and other possible electrically charged parts, and not to take and put things in the electrophoresis tank, if necessary, the power should be disconnected first to avoid electric shock. At the same time, it is required that the instrument must have a good grounding end to prevent leakage.
b. After the instrument is energized, do not temporarily add or dial the output wire plug to prevent short-circuit phenomenon, although the instrument is equipped with an internal fuse, the short-circuit phenomenon may still lead to instrument damage.
c. Since the resistance value of different media supports is different, the electric flow through the electrophoresis is also different, and its swimming speed and the time required to swim to the endpoint are also different, so the electrophoresis of different media supports should not be carried out on the same electrophoresis instrument at the same time.
d. When the total current does not exceed the rated current of the instrument (maximum current range), it is possible to use multiple slots in association, but be careful not to overload, otherwise it will easily affect the instrument’s life.
e. Under some special circumstances, especially when the instrument needs to check the electrophoresis input, it is allowed to start the machine at no load in the state of voltage stabilization, but in the state of current stabilization, the load must be connected before starting the machine, otherwise the voltmeter pointer will jump significantly, which will easily cause unnecessary man-made machine damage.
f. If abnormal phenomena are found during use, such as loud noise, discharge, or abnormal odor, the power shall be immediately cut off and overhauled to avoid accidents.
Common troubleshooting of electrophoresis apparatus
A. The output of the electrophoresis apparatus does not reach the set value
The output value state of the electrophoresis apparatus follows "Ohm's law": voltage U=current I×(electrophoresis tank) resistance R
If the resistance R is relatively constant, any one parameter of U, I, and P (power P=current I×voltage U) is constant, other parameters are also constant; and if any one parameter changes, other parameters also change proportionally.
If the output voltage U of the electrophoresis instrument does not reach the preset value, you should first observe whether I or P has been constant, or has reached the maximum I or P specified by the electrophoresis instrument (JY electrophoresis instruments all have clear indicator signs). If the limit value has not yet been reached, turn up the setting of already constant I or P (to the limit value if necessary) to be able to increase the voltage output.
If the current I of the electrophoresis instrument does not reach the preset value, adjust the voltage U or power P. If the power P of the electrophoresis instrument does not reach the preset value, adjust the voltage U or current I.
B. Computer-controlled electrophoresis instrument over-voltage alarm
a. Check whether it is used with no load.
b. Whether the electrophoresis tank is not filled with buffer.
c. Whether the platinum wire of the electrophoresis tank is broken.
C. Overcurrent protection
a. Whether there is a short circuit in the electrophoresis tank.
b. Whether the buffer is wrongly selected.
D. Leakage protection
a. Whether there is liquid spilled into the inside of the instrument or on the output interface.
b. Whether there is a lot of dust falling into the inside of the instrument.
How to buy electrophoresis apparatus?
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