PT full wavelength enzyme-linked immunosorbent assay (ELISA) readerIt is a high-precision analytical instrument that integrates multiple detection modes such as UV visible absorption, fluorescence, chemiluminescence, etc. It is widely used in fields such as life sciences, clinical diagnosis, drug development, and food safety. Its core advantages lie in wide wavelength coverage (usually 190-1000nm) and multifunctional detection capabilities, which can meet diverse needs from basic scientific research to industrial production. It is based on the law of light absorption (Bill Lambert law), emitting light of a specific wavelength through a light source. After being absorbed by the sample, the detector measures the remaining light intensity and calculates the absorbance (A) to determine the concentration of the target molecule.

　　PT full wavelength enzyme-linked immunosorbent assay (ELISA) readerThe operation needs to be combined with experimental requirements (such as detection mode, sample type) for parameter setting and process control. The core steps include "preparation setting detection data processing". The following is the general operation process:

1、 Preparation before operation

Sample and reagent preparation

Ensure that the samples to be tested (such as ELISA plates, nucleic acid solutions, cell suspensions, etc.) have been processed according to the experimental protocol, and that there are no bubbles or droplets hanging on the walls of the microplate (if there are bubbles, gently tap the plate wall to remove them; for hanging droplets, use paper towels to dry the edges).

Prepare blank controls, standard samples, and quality control samples, and arrange them in a predetermined layout on a microplate (it is recommended to record the hole layout to avoid confusion).

Instrument and environmental inspection

Confirm that the power cord of the enzyme-linked immunosorbent assay (ELISA) reader is connected properly. Before starting up, check if the sample chamber is clean (without dust or liquid residue) and wipe it with a dust-free cloth if necessary.

If the experiment requires temperature control (such as incubation at 37 ℃) or shaking, confirm in advance that the relevant functions of the instrument are normal (such as whether the temperature control module can reach the set temperature).

Turn on the computer and power of the enzyme-linked immunosorbent assay (ELISA) reader, start the supporting operating software, and wait for the instrument to complete its self-test (usually showing a "ready" status).

2、 Detection method setting

Select detection mode

Select the corresponding detection mode in the software based on the type of experiment:

Absorption light detection (such as ELISA, nucleic acid quantification): The detection wavelength (such as 450nm main wavelength, 630nm reference wavelength) and bandwidth (default 1-5nm) need to be set.

Fluorescence detection: It is necessary to set the excitation wavelength (such as 485nm), emission wavelength (such as 535nm), and select the top/bottom reading (liquid samples usually choose the top, cell cultures can choose the bottom).

Chemiluminescence detection: No wavelength setting is required, simply select the "chemiluminescence" mode, and be careful to turn off the sample chamber light source to reduce interference.

Dynamics detection: Additional detection time intervals (such as reading every 10 seconds), total detection duration (such as 30 minutes), and temperature control (if constant temperature reaction is required) need to be set.

Set microplate parameters

Select the specifications of the microplate (such as 96 well plate, 384 well plate) and define the detection range (such as "full plate detection" or specifying specific well positions, such as A1-H12).

If edge effects or partial hole positions need to be excluded, mark "not detected" holes in the software.

Auxiliary function settings

Temperature control: If the reaction requires constant temperature (such as enzymatic reaction at 37 ℃), set the target temperature (usually with an error of ± 0.1 ℃) and wait for the instrument to preheat to the set temperature.

Vibration: When mixing the sample before testing, set the vibration mode (such as linear vibration), speed (such as 500rpm), and time (such as 10 seconds).

Reference wavelength correction (absorption light detection): Turn on "dual wavelength detection" and subtract background interference (such as microplate scratches and bubbles) from the reference wavelength (such as 630nm) to improve data accuracy.

3、 Sample testing

Loading microplate

Gently place the microplate onto the tray in the sample chamber, ensuring that the edge of the plate aligns with the positioning groove of the tray (to avoid reading errors caused by positional deviation), and close the sample chamber door.

Startup detection

Click "Start Detection" in the software, and the instrument will automatically run according to the preset method. The screen will display the detection progress in real time (such as completed hole positions and current readings).

During the testing process, avoid touching the instrument or opening the sample chamber door to prevent interference with the stability of the optical path.

4、 Data Processing and Export

View real-time results

After the detection is completed, the software automatically generates raw data (such as absorbance values, fluorescence intensity) and displays it in the form of tables or heat maps.

You can view single well data, average values (such as the mean of standard replicate wells), or generate standard curves (if standard concentration is set).

data analysis

Process data according to experimental requirements:

Quantitative analysis (such as ELISA): The software automatically calculates the unknown sample concentration by fitting the standard curve (linear, four parameter fitting, etc.) with the standard concentration and corresponding signal values.

Ratio calculation (such as nucleic acid purity): Automatically calculate the 260/280nm and 260/230nm ratios in absorption light detection to evaluate sample purity.

Dynamics curve: Generate a "signal value time" curve, which can calculate parameters such as reaction rate and half-life.

Data export and saving

Save the data in the software's default format (such as. csv,. txt), or export it to Excel or PDF format for report writing.

It is recommended to save the detection method template at the same time (the same experiment can be directly called next time without the need for repeated settings).

5、 Organize after operation

Turn off the instrument

After the testing is completed, remove the microplate and clean the sample chamber (if there is any liquid leakage, wipe it promptly).

Turn off the enzyme-linked immunosorbent assay software, instrument power, and computer power in sequence.

Recording and Cleaning

Record experimental information (such as testing date, instrument model, method parameters) for easy data traceability.

Regularly clean the sample room and trays (wipe with 75% alcohol to avoid residual corrosive reagents), and cover them with dust covers when not in use for a long time.