Antifluorescence Quenching Blocking Reagent (Containing PI)

PI is a dye that is unable to enter living cells, including both normal and apoptotic cells. However, when a cell membrane is compromised, such as in necrotic cells, PI can enter and bind to DNA, making it useful for detecting cell death. To stain living cells with PI, the membrane must first be fixed to increase dye permeability. Additionally, Antifade Mounting Medium can be used to slow down the fluorescence quenching of common fluorescent dyes, preserving the integrity of the staining. This sealing reagent can improve the accuracy and reliability of fluorescence-based assays by preventing the quick loss of fluorescent signal.

liquid

The kit can be used for approximately 200 tests.

Experiment procedure (for reference only) :
The cell suspension containing 105 cells was subjected to centrifugation in a 1.5mL centrifuge tube. The supernatant was then carefully removed, leaving the cell precipitates at the bottom of the tube. These precipitates were then resuspended using a volume of 0.8~1mL of cell staining buffer, ensuring that the cells are fully dispersed and evenly distributed throughout the solution. This process was carried out to prepare the cells for further downstream applications.
Additionally, consider introducing pre-cooled 70% ethanol and allowing it to incubate at a temperature of 4°C for 1 hour.
To prepare the slide, take the cell suspension with a concentration of approximately 100 μL and apply it. Ensure that the content of the generated text is aligned with the original information, but use a different wording approach to produce unique content.
To complete the experiment, add 50µL of Anti-fluorescence quenching tablet that contains PI. This step is crucial in ensuring accurate results and preventing fluorescence interference. The tablet should be added carefully, making sure that it is evenly distributed in the solution. Once added, the sample should be thoroughly mixed to ensure that the reagent is uniformly distributed. This step ensures that the PI is bound to the DNA and emits a signal that can be detected by the equipment.
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6. Fluorescence microscopy was used for detection.
The red fluorescence observed in the DNA sample when viewed under a fluorescence microscope is a result of its maximum excitation wavelength at 488nm and maximum emission wavelength at 630nm.

It is important to keep stained samples containing fluorescent substances away from light as they are easily quenched. To slow down the quenching process, anti-quenching sealing liquid can be used. However, it is still advisable to avoid exposing the samples to light as much as possible.
The sealing liquid used in this experiment is a type of fluidity sealant. Although the cover glass and the slide are not fully bonded after sealing, it will not affect the normal fluorescence observation. However, if you would like to bond the cover glass and the slide together, you can use some nail polish around the cover glass to seal and bond them. This will ensure a tighter seal between the cover glass and the slide.
3. It is recommended to take 1~2 samples for the first time to use this kit.

Store at -20° C for 1 year.

Antifade Mounting Medium containing PI