Rhodamine 123 is a fluorescent dye that can selectively stain live cell mitochondria through the cell membrane, showing yellow-green fluorescence. It is widely used to detect mitochondrial membrane potential and is also commonly used to detect apoptosis. It can quickly pass through cell membranes, can be captured by active mitochondria in just a few minutes, and does not have any toxicity to cells.
Rh123 is used to stain many kinds of cells, including plant cells and bacteria. Due to the correlation between the amount of ATP in the cell and the fluorescence intensity of Rh123, Rh123 was used to detect ATP in the cell. Rh123 is also used in cancer research.
Rhodamine 123 dyeing procedure:
(1) Dissolve 0.4mgRh123 in 1mL DMSO to prepare 1mMRh123-DMSO solution.
(2) Prepare the cells with a glass slide. The number of cells should be 5 × 104 ~ 5 × 105 cells / mL.
(3) Incubate the slide and wash the cells with PBS or Hank's solution.
(4) Dilute 1mM Rh123 solution with culture medium to prepare 1 ~ 20μM Rh123 buffer.
(5) Add Rh123 buffer to the slide and incubate at 37 ° C for 30 minutes to 1 hour.
(6) Remove the Rh123 buffer and wash the cells with the culture medium (after washing the cells for fixation, add 10% formalin buffer and incubate for 15-20 minutes, then wash with PBS).
(7) Observe the cells with a fluorescence microscope with a fluorescein filter.
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