Method for performing biological mark detection by atomic force microscope
An atomic force microscope and biomarker technology, applied in biological testing, scanning probe microscopy, nanotechnology for sensing, etc., can solve problems such as limitations in the development of new alternative technologies, and achieve overall shortened time and low price. , handle simple effects
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[0031] 1. Preparation of gold nanoparticles
[0032] 1. Boil gold
[0033] Soak the triangular burner in sulfuric acid for 24 hours before use, take it out and wash it 10 times with water, 8 times with distilled water, and then 6 times with double distilled water. Take 100ml of double-distilled water from the washed triangular burner, first boil it in a microwave oven, then put it on a heating magnetic stirrer, adjust the rotor so that there is no liquid splashing, and the rotation is stable, add 1ml of chloroauric acid, 1.5ml of disodium citrate, of which Add chloroauric acid first, then add disodium citrate, 1.5ml should be added together. Observe the color change, colorless-black-wine red, until the color is stable to wine red, cool, store in a brown bottle at 4°C.
[0034] 2. Adjust the PH value
[0035] Use a small Erlenmeyer flask (for nano-gold) to take 15ml of boiled gold, adjust the pH value with 1M K2CO3, add 70μl K2CO3 pH value between 8.5 and 9.0.
[0036] 3. D...
Embodiment 1
[0052] Put the 15mm circular sample carrier glass slide into the self-made washing solution (including 3% sulfuric acid, 3% AES, 0.4% sodium hydroxide, 1.2-1.5% sodium chloride), ultrasonically wash for 10min, take out the nitrogen gas and blow dry, polylysine Acid treatment 5min, natural drying. Stick the glass slide on a circular patch with a diameter of 15mm dedicated to AFM, and scan the sample with the tapping mode of AFM. The resulting image see Figure 1A : The background of the sample is uniform, the fluctuation is less than 1nm, the surface has a porous structure, which is easy for the sample to be fixed by physical adsorption, and the hardness of each area is moderate.
[0053] Take 50 μl of cooked nano-gold solution, evenly spread it on the surface of the glass slide treated with polylysine, incubate at 37°C for 30 minutes, wash 5 times with 1×PBS, 1 minute each time, wash 3 times with deionized water, 3 minutes each time, Blow dry with nitrogen. Stick the glass s...
Embodiment 2
[0056] Cloning and inducible engineering strain BL-21 with measles virus nucleoprotein (BL-21-30a-MVn strain, from the Infection Department of Alien Diseases, Chinese Academy of Inspection and Quarantine Sciences), using a pipette tip to pick LB solid medium Transfer a single colony into LB liquid medium, shake overnight in a constant temperature shaker (37°C, 265rad / min); take 1ml of the bacterial solution into a 1.5ml EP tube, centrifuge at 3000rpm for 10min, discard the supernatant, and then use 1ml of 1× After the bacteria were suspended in PBS and precipitated, centrifuge at 3000 rpm for 10 min, wash three times according to the above steps, and finally suspend the washed bacteria cells in 500 μl 1×PBS. Take 50 μl of the bacterial solution and apply it evenly on two 15mm round glass slides treated with polylysine, incubate at a constant temperature of 37°C for 30 minutes, absorb the bacterial solution on the surface of the slides with filter paper, wash one of the samples ...
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