Experimental principle Mitochondria are an important organelle in cells and a place where cells breathe. The energy required for the activities of the cells is mainly provided by mitochondrial respiration. In vivo staining is a staining method that uses non-toxic or less toxic stains to truly reveal certain structures in living cells with little impact on cell life. Jenner Green B is a specific living body stain for the line body. The cytochrome oxidase in the mitochondria keeps the dye in a oxidized state, which is blue-green, and the dye is reduced in the surrounding cytoplasm to become a colorless state. Experimental reagent 1. 1/300 Jenus Green B dye solution 2. Ringer's solution (for mammals) Laboratory equipment Microscope 2. A set of surgical equipment 3. Dissection disk 4. Small plate 5. Slide 6. Cover slip 7. Absorbent paper 8. 10ml syringe 9. Straw Experimental Materials Electron micrograph of a rabbit, mitochondria. Experimental procedure 1. Liver staining of rabbit liver cell mitochondria 1) The rabbit was sacrificed by air embolization, placed in an anatomical tray, and the abdominal cavity was quickly opened. 2) Take a small piece (2~3mm3) of the liver tissue with thin edge of the rabbit liver, put it into the dish containing Ringer's solution, wash the blood (lightly pressed with tweezers), and suck the Ringer's solution with a straw to make it in the plate. Add 1/300 Jenus Green B dye solution to expose the upper surface of the tissue block to the outside of the dye solution, so that the mitochondrial enzyme system in the cell can be fully oxidized, which is beneficial to maintain the oxidation state of the dye and color the mitochondria. . When the edge of the tissue block is dyed in a basket, it usually takes 30 minutes. 3) After staining, the tissue block is moved onto the slide, and the tissue block is broken with tweezers, and some cells or cell populations are detached from the tissue block. Remove the slightly larger tissue block and leave the free cells or cell population on the slide. Add a drop of Ringer's solution and cover the cover to remove excess water. 2. Results 1) Microscopic observation, many mitochondria in the liver cytoplasm are stained blue-green and granular. 2) Light microscopy section observation of mitochondria The rabbit liver cells stained with Jenus Green B were sectioned by light microscopy, and the mitochondria in the liver cells were blue-green particles. 3) Electron microscopic observation of mitochondria The morphology and number of mitochondria are different in different cells. Mitochondria have a variety of shapes, such as round, oval, dumbbell and rod. The number of mitochondria is related to the cell type and the physiological state of the cells, and the mitochondria are mostly concentrated in areas where the physiological functions of the cells are strong. The number and distribution of mitochondrial ridges is varied. It is generally aligned perpendicular to the long axis of the mitochondria, but ridges arranged parallel to the long axis of the mitochondria are also visible. The transverse section of the ridge is saclike or tubular. The number of ridges is strongly dependent on the strength of cellular ventilatory function. Shanghai J.Shine Co.,Ltd , https://www.jshinechem.com