Fixed liquid special for electron microscopy sample preparation of biological tissue sample and preparation method of fixed liquid
A technology of biological tissue and fixative, which is applied in the field of special fixative for electron microscope sample preparation of biological tissue samples and its preparation, which can solve the problems of poor preservation of microtubules and achieve the effect that bacteria are not easy to grow
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[0019] A method for preparing a special fixative for electron microscope sample preparation of biological tissue samples, comprising the following steps:
[0020] Step 1: Dissolve 1.0-3.0g of paraformaldehyde powder in 20.0-30.0ml of distilled water, raise the temperature to below 65°C and keep stirring;
[0021] Step 2: Add 1.0-3.0ml of 1N sodium hydroxide while stirring. After the solution is transparent, add 3.2-6.6ml of 5% glutaraldehyde after cooling;
[0022] Step 3, add cacodylate buffer solution to the mixed solution to make the volume reach 50ml; the preparation method of cacodylate buffer solution is: prepare 4.0-5.2g sodium cacodylate, add distilled water to 100ml; 0.2N 1 - 20ml of hydrochloric acid, mix and dilute sodium cacodylate solution with 0.2N 1-20ml hydrochloric acid to 200ml to obtain cacodylate buffer solutions with different pH;
[0023] Step 4, finally add 22.0-26.2 mg of anhydrous calcium chloride, adjust the pH of the mixed solution to 6.8-7.5 to obt...
Embodiment 1
[0025] Dissolve 1.2g of paraformaldehyde powder in 20.0ml of distilled water, raise the temperature to below 65°C and keep stirring; add 1.0ml of 1N sodium hydroxide while stirring, after the solution is transparent, add 5% glutaraldehyde 3.2 after cooling ml; add cacodylic acid buffer solution to the mixed solution to make the volume reach 50.0ml;
[0026] The preparation method of cacodylate buffer solution is as follows: prepare 4.0g sodium cacodylate, add distilled water to 100ml; 0.2N1-20ml hydrochloric acid, mix and dilute sodium cacodylate solution with 0.2N 1-20ml hydrochloric acid to 200ml, Cacodylate buffer solution with different pH was obtained, see Table 1 for details;
[0027] pH
6.8
7.0
7.2
7.4
7.6
7.8
Sodium cacodylate ml
50
50
50
50
50
50
0.2N hydrochloric acidml
18.3
13.3
9.3
6.3
4.2
2.7
Add double distilled water to ml
200
200
200
200
200
200
[0028] ...
Embodiment 2
[0031] Dissolve 2.0g of paraformaldehyde powder in 25.0ml of distilled water, raise the temperature to below 65°C and keep stirring; while stirring, add 3.0ml of 1N sodium hydroxide, after the solution is transparent, cool and add 5% glutaraldehyde 5.0 ml; add cacodylate buffer solution to the mixed solution to make the volume reach 50.0ml; the preparation method of cacodylate buffer solution is: prepare 4.0g sodium cacodylate, add distilled water to 100ml; 0.2N 1-20ml For hydrochloric acid, mix sodium cacodylate solution with 0.2N 1-20ml hydrochloric acid and dilute it to 200ml to obtain cacodylate buffer solutions with different pHs, see Table 2 for details;
[0032] pH
6.4
6.6
6.8
7.0
7.2
7.4
Sodium cacodylate ml
50
50
50
50
50
50
0.2N hydrochloric acidml
18.3
13.3
9.3
6.3
4.2
2.7
Add double distilled water to ml
200
200
200
200
200
200
[0033] Table II
[0034] Step...
PUM
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