Method for analyzing cartilage tissue
A cartilage tissue and cartilage matrix technology, applied in the field of cartilage tissue analysis, can solve the problems of cartilage tissue sample consumption, one-sided cartilage information, small slice area and thickness, etc., to achieve real-time monitoring of cartilage repair effect, comprehensive and accurate cartilage information, Avoid wasteful effects
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Embodiment 1
[0117] (1) Using a two-photon confocal microscope to perform an autofluorescence scanning experiment on the cartilage matrix in the cartilage tissue of the animal model, and obtain the autofluorescence excitation conditions of the cartilage matrix.
[0118] 1) Take fresh knee joints of Sprague Dawley (SD) rats, dissect the knee joints, and remove soft tissues such as muscles, fascia and fat around the knee joints. The surface of the knee joint was washed with normal saline, and the subchondral bone was trimmed to obtain a flat cartilage tissue sample of the SD rat knee joint, which was used for detection by two-photon confocal microscopy. Among them, care should be taken to protect the articular surface from damage during the process.
[0119]2) Transfer the cartilage tissue sample of the SD rat knee joint into a confocal vessel, with the part to be tested facing downward, and perform fluorescence detection experiments on it using a Leica SP8 inverted two-photon confocal micro...
Embodiment 2
[0145] In this example, the normal cartilage tissue samples scanned by the confocal microscope in Example 1 were subjected to conventional fluorescent staining of cell nuclei and cytoskeleton, and the stained normal cartilage tissue samples were fluorescently stained using a Leica SP8 two-photon confocal microscope. The scanning process obtains the fluorescence images of the nuclei and cytoskeleton at different levels in the normal cartilage tissue sample, so as to further observe the distribution and shape of the chondrocytes in the sample, and further provide information for the analysis of the cartilage tissue.
[0146] The specific steps of sample preparation are as follows: sample fixation: 10% neutral formaldehyde fixes the sample for 12 hours, and rinses with running water for 24 hours. Stain with 140 nm rhodamine-phalloidin for 12 hours at 4°C in the dark. Phosphate Buffered Saline (Phosphate Buffered Saline, pH=7.4) solution was soaked for 24 hours, and the medium was...
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