Method for marking cells by using protein fluorescent probes
A fluorescent probe and cell technology, applied in animal cells, vertebrate cells, artificial cell constructs, etc., can solve problems such as high cost, limited development and research, and large cell damage
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Embodiment 1
[0063] Example 1: Fluorescence of Probe Suspension
[0064] Take 0.096g of graft copolymer with a graft degree of 30 and dissolve it in a centrifuge tube filled with 5mL of sterilized water, and disperse the polymer with an ultrasonic cleaner. During ultrasonic dispersion, the ultrasonic cleaner stops for 10 minutes every 20 minutes of work. Ultrasonic time is about 7h. Then, under a fluorescent microscope, ultraviolet excitation was used to observe the fluorescence of the copolymer suspension, such as figure 1 As shown, the particle size of the polymer particles can be dispersed to about 500nm, and the blue fluorescence intensity is very strong, which lays the foundation for the co-incubation of the culture liquid suspension and the cells.
Embodiment 2
[0065] Example 2: Fluorescence in cells when graft copolymer was co-cultured with CHL cell tumor for 6 hours
[0066] CHL cells were co-cultured with high glucose medium. When the cells are passaged, the cells are made into a cell suspension and distributed to six-well plates covered with coverslips. in CO 2 After culturing in the incubator for 24 hours, the medium was changed, and the new culture medium was a suspension in which the copolymer was dissolved. After the cells and the culture medium suspension continued to co-incubate for 6 hours, the culture medium was discarded, washed with PBS 3 times for about 5 min each time, and shaken on a shaker. Take out the cover glass and put it on the glass slide and put another cover glass on it. Under the fluorescence microscope, the excitation wavelength is ultraviolet excitation, the range is EX=330-380nm, the emission wavelength range is EM=435-485nm, and the exposure time is 15.505ms, observe the cell condition. Such as im...
Embodiment 3
[0067] Example 3: Fluorescence in the cells when the graft copolymer was co-cultured with CHL cells for 12 hours
[0068] The graft copolymer was co-cultured with CHL cells for 12 hours, and the experimental process and treatment methods were the same as in Example 3. Such as Figure 4 As shown, the graft copolymer of small particles enters the cell, and the way that the extremely small nano-scale substance enters the cell is mainly permeation and diffusion. From the type of CHL cells and the way of feeding, we know that the graft copolymers of small particles in the cells are mainly carried out through the endocytosis of the cells, and the fluorescence of the graft copolymers begins to appear in the cells.
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