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Cat. No.: AOPIA-100 (for Set A, 100T)

Cat. No.: AOPIA-500 (for Set A, 500T)

Cat. No.: AOPIB-100 (for Set B, 100T)

Cat. No.: AOPIB-500 (for Set B, 500T)

Cat. No.: AOPIC-100 (for Set C, 100T)

Cat. No.: AOPIC-500 (for Set C, 500T)

Description

AO/PI Double Staining Kit for Cell Counter, also known as AO/PI dye or AO/PI staining solution, is a reagent kit designed for quick and convenient detection of cell viability. It combines acridine orange (AO) staining solution and propidium iodide (PI) staining solution in an optimized ratio, based on dual staining of cell nuclei with DNA probes.

This product line includes Set A, Set B, and Set C, suitable for fluorescence cell counters with different fluorescence intensity requirements. The fluorescence strength ranges from weak in Set A to strong in Set C. For first-time purchasers, it is recommended to start with Set B. Typically, parameters of the fluorescence cell counter can be adjusted to accommodate different fluorescence intensities. If these three sets are not suitable for a specific fluorescence cell counter, you can opt for the AO/PI Double Staining Kit (Test Version, Cell Counter Specific) (C2021) to test the appropriate amount of fluorescent dye, and if necessary, contact us for customization.

AO is a classic, highly sensitive fluorescent dye that can stain DNA and RNA. It is membrane-permeable, allowing it to pass through cell membranes and stain DNA and RNA. AO binds to nucleic acids in two ways: intercalation and electrostatic attraction. Intercalation occurs when AO inserts itself between the base pairs of double-stranded DNA, emitting green fluorescence at an emission peak of 530 nm when excited. Electrostatic attraction happens when the positively charged AO binds to the negatively charged phosphate groups of RNA or single-stranded DNA, emitting red fluorescence at an emission peak of 640 nm, with small amounts emitting orange or red fluorescence. In this kit, AO binds to nucleic acids through intercalation, resulting in green fluorescence upon excitation. After AO staining, the nuclei of normal cells exhibit uniform green or yellow-green fluorescence. In apoptotic cells, due to chromatin condensation or fragmentation into uneven-sized fragments forming apoptotic bodies, AO stains them with dense yellow-green fluorescence or yellow-green granular fragments. In necrotic cells, the yellow fluorescence weakens or disappears.

PI is a non-cell membrane-permeable fluorescent dye. After binding to DNA, it has a maximum excitation wavelength of 535 nm and a maximum emission wavelength of 617 nm. It cannot penetrate intact cell membranes and can only reach the nuclei of necrotic cells due to their compromised membrane integrity, binding to the DNA double helix to form PI-DNA complexes that emit red fluorescence. Therefore, PI only stains dead cells and is used to distinguish normal cells from necrotic cells. It is important to note that apoptotic cells cannot be stained by PI due to their intact cell membranes, only staining during the late stages of apoptosis when secondary necrosis occurs.

Features

• This series of kits is easy to use and provides quick detection. The kits offer ready-to-use AO/PI double staining reagents. To use, simply mix the reagents with the cell samples in a 1:1 ratio. This eliminates the cumbersome process of preparing and diluting various solutions, reducing potential errors during preparation and dilution and improving the accuracy of detection. The entire staining process can be completed within 5 minutes.
• The series offers products with different detection sensitivities for users to choose from, suitable for various cell counters. Different brands of fluorescence cell counters have varying fluorescence detection parameters, which can be adjusted accordingly. Therefore, this series of products is divided into three sets based on staining intensity: Set A, Set B, and Set C, with green fluorescence staining intensity increasing sequentially. Users can select the appropriate set according to the characteristics of the cell counter used in their experiments.

Storage

• Store at 4ºC in the dark, valid for one year.
• Store at -20ºC in the dark, it can be stored for a longer period.
• Store at room temperature in the dark, valid for one month.

Precautions

• Fluorescent dyes are prone to quenching. Please keep them in the dark to slow down fluorescence quenching.
• This product is for scientific research purposes only and must be used by professionals. It is not intended for clinical diagnosis or treatment, food, or drug usage, and should not be stored in a typical household.