CytoFLEX Ready to Use Daily QC Quality Control Fluorescent Microsphere Instrument Calibration Sample

Flow cytometry is an important cell analysis technique that can be used for cell counting, phenotype analysis, and cell sorting. Quality control is crucial before conducting flow cytometry to ensure the accuracy and reliability of the experiment. This article will provide a detailed introduction to the quality control methods of flow cytometry.

1. Fluid system calibration

The fluid system includes flow meters and pressure sensors. Firstly, calibrate the flow meter and pressure sensor by measuring the actual flow rate and pressure. Inject standard fluid into the flowmeter, measure the flow rate, and calibrate it. Inject standard pressure into the pressure sensor and measure the output value, then calibrate.

2. Laser system calibration

The purpose of calibrating a laser system is to determine the position of the laser beam in order to obtain accurate optical readings in the sample container. Fix the calibration bead in the sample tube of the flow cytometer, irradiate the calibration bead with laser and determine the position of the laser beam.

3. Optical system calibration

Calibrating the optical system is key to ensuring accurate optical readings during sample analysis. The calibration optical system includes a detector and a synchronizer. The detector is responsible for measuring the number and size of immunostaining particles in the sample. The synchronizer is responsible for ensuring that the detector measures the sample at the correct frequency.

4. Standard particle calibration

The final step in calibrating a flow cytometer is to standardize the sample using a standard particle calibrator. Use spacious sample tubes to test standard particles at different concentrations. After calibrating the standard curve, the flow cytometer can accurately calculate the number and characteristics of specific immunostaining particles in the sample.

Flow cytometer calibration

1. Clean the sample chamber of the flow cytometer and check if the laser and optical system are working properly.

2. Use particle standard substances to check the accuracy of the scattering, fluorescence, and time parameters of the cell analyzer.

3. Clean and calibrate the liquid system of the flow cytometer to ensure accurate control of flow rate and sample size.

Quality Control of Flow Cytometer Performance

a. Regularly check the performance parameters of the flow cytometer, including scattering sensitivity, fluorescence detection sensitivity, and time resolution.

b. Use calibration particles to verify the accuracy and consistency of flow rate,

c. According to the experimental requirements, use appropriate quality inspection particles to adjust the resolution and intensity

Sample preparation

1. Uniformly suspend cell samples to avoid cell aggregation and deposition. This can be achieved through centrifugation and resuspension of the sample.

2. Use reliable cell staining methods to ensure accuracy and consistency of staining.

3. Adjust the concentration of the sample to avoid excessive focusing and loss of signal due to sparse cells.

Quality control of sample purity

a. Use appropriate negative and positive controls to evaluate the purity of staining. For example, using unstained cells as negative controls and cells with known positive markers as positive controls

b. Regularly check the consistency of staining results to ensure that negative control cells have no positive markers and positive control cells have expected positive markers

CytoFLEX Ready to Use Daily QC Quality Control Fluorescent Microsphere Instrument Calibration SampleChinese introduction

CytoFLEX ready to use daily QC fluorescent microspheres are a suspension of 3 μ m fluorescent microspheres, which can be used for optical alignment of flow cytometers and daily validation of flow systems. This product is provided in pre diluted ready to use form, with fluorescence emission wavelengths between 355nm and 800nm when excited at the following wavelengths: 355nm, 375nm, 405nm, 488nm, 561nm, and 638nm. The mixture is suspended in an aqueous medium containing surfactants and stabilizers. C65719 replaces CytoFLEX daily QC fluorescent ball B53230 and is suitable for CytoFLEX cell analyzer platform.

CytoFLEX Ready to Use Daily QC Quality Control Fluorescent Microsphere Instrument Calibration SampleEnglish introduction

CytoFLEX Ready to Use Daily QC Fluorospheres is a suspension of 3 µm fluorescent microspheres, which may be used for daily verification of the flow cytometer's optical alignment and fluidics system. This product is provided in a pre-diluted, ready-to-use format, with a fluorescence emission between 355 nm to 800 nm when excited at: 355 nm, 375 nm, 405 nm, 488 nm, 561 nm, and 638 nm. The mixture is suspended in an aqueous medium containing surfactants and stabilizers. Click here for CytoFLEX Ready to Use Daily QC Fluorospheres Instructions for Use. This product replaces the CytoFLEX Daily QC Fluorospheres, B53230, and should be used with the CytoFLEX cytometer platform.

CytoFLEX Ready to Use Daily QC Quality Control Fluorescent Microsphere Instrument Calibration Sampleoperating procedure

1. Use the filter set recommended by the manufacturer for detecting the appropriate fluorescence parameters (refer to the CytoFLEX Platform IFU).

2. Ensure that the target value file for the specific lot of CytoFLEX Ready to Use Daily QC Fluorospheres has been loaded into the flow cytometer software. Refer to your CytoFLEX Platform IFU for instructions on importing your lot-specific target value file.

3. Vigorously mix the CytoFLEX Ready to Use Daily QC Fluorospheres vial using a vortex mixer for 2-3 seconds.

4. Add:

4. 10 drops of fluorospheres to a tube if using Semi Automatic (tube) Sample Injection Mode.

b. 3-4 drops of fluorospheres to a well for either Standard 96-well plate or Deep-well plate for the plate loader injection mode.

5. Run the Quality Control (QC) protocol per the instructions in your CytoFLEX Platform (IFU).

CytoFLEX Ready to Use Daily QC Quality Control Fluorescent Microsphere Instrument Calibration SamplePrecautions

1. The product is a ready to use product that does not require additional preparation and can be used directly.

If packaged and sealed, use up within 5 days.

3. Fluorescent microspheres will naturally settle after prolonged standing. Ensure that the fluorescent ball is thoroughly resuspended and shaken before use.

4. Do not use fluorescent balls that have exceeded the expiration date on the sample bottle label.

5. The product should be refrigerated and not frozen.

When handling this reagent, please use Good Laboratory Practice (GLP).

7. This reagent contains preservatives. If skin or eye contact occurs, please rinse thoroughly with water in a timely manner.

The purpose of the flow cytometer calibration report is to record detailed information about all flow cytometer calibration processes. The report should include the following information:

1. Date and time

2. Fluid system calibration data

3. Laser system calibration data

4. Optical system calibration data

5. Standard particle calibration data

6. Calibration bead calibration data

7. Any problems or abnormal situations discovered during the calibration process

8. Name and signature of the technical personnel performing the calibration