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RWD How to Achieve Precise Cell Counting with AOPI Staining

RWD How to Achieve Precise Cell Counting with AOPI Staining
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RWD How to Achieve Precise Cell Counting with AOPI Staining - 1

How to Achieve Precise Cell Counting with AO/PI Staining? Cell concentration and viability detection is one of the most basic cell culture processes, and cell staining is required to perform cell counting and viability assays. The commonly used stains include trypan blue and acridine orange/propidium iodide (AO/PI). Trypan Blue Staining Trypan blue is a stain commonly used to distinguish live cells from dead cells. Since the membrane of live cells is selectively permeable and can block trypan blue, live cells will not be stained. The membrane of dead cells is more permeable, so dead cells can be stained blue by trypan blue. If trypan blue staining is adopted in the experiment, the viability and concentration of the cell sample can be quickly calculated by observing and counting the stained and unstained cells under the microscope. For more accurate cell-counting, trypan blue is not an ideal method. Since it usually stains the debris of primary cells and residual red blood cells in PBMCs. Besides, there is no standard for the preparation of dye solution, so that different solution may lead to different counting results. In addition, trypan blue is toxic to cultured cells, and it may destroy the cells if staining for a long time. However, in practical applications, the actual results will be different due to the different cell types, staining conditions and parameters of detection instruments, in that case, it is hard to guarantee stable staining results. trypan blue staining -1-

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RWD How to Achieve Precise Cell Counting with AOPI Staining - 2

AO/PI Staining The fluorescent dye AO can penetrate the cell membrane and stain DNA and RNA, respectively, producing green fluorescence(DNA) and orange fluorescence(RNA). The fluorescent dye PI is a nuclear staining reagent that can stain DNA, which is commonly used for the detection of apoptosis. It is an analog of ethidium bromide, and emits red fluorescence after embedding itself in double-stranded DNA. PI cannot pass through the living cell membrane, but can only pass through the broken cell membrane to stain the nucleus. Since debris, impurities, or mammalian red blood cells do not...

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RWD How to Achieve Precise Cell Counting with AOPI Staining - 3

well, then incubate for 5 min (final concentration: PI, 25 μg/mL; AO, 5 μg/mL); 3.Gently mix the solution, then pipette 10 μL of stained cells into the cell counting plate for analysis. RWD Automated Cell Counter is recommended. This device can accurately and rapidly distinguish living cells from dead cells stained with AO/PI dual fluorescence by one-click counting. cell counting using RWD Automated Cell Counter By Counter (https://www.rwdstco.com/product-item/automatedcellcounter/) can deliver accurate and stable cell staining counting. It is widely used in multiple application scenarios...

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