Fast fluorescent staining method of cruciferae crop pollen
A cruciferous crop, fluorescent dyeing technology, applied in the preparation and sampling of scientific instruments, samples for testing, etc., can solve the problems of time-consuming, cumbersome, labor-intensive, etc., and achieve obvious contrast, good repeatability, and reduced cytoplasmic and nuclear staining. Effects of Environmental Pollution
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Embodiment 1
[0025] Example 1: Fluorescence dyeing method of mature pollen of rape blossoms
[0026] 1. Staining solution preparation: first dissolve Hoechst33258 fluorescent dye and sucrose in phosphate buffer saline (PBS, pH=7.4), prepare Hoechst33258 concentration 20μg / L, sucrose concentration 10wt% solution 1, and then prepare 60% ethanol ( v / v), ethyl acetic acid 10% (v / v), glycerol 30% (v / v) solution two, mix solution one and solution two in a volume ratio of 1:1 to get the dyeing solution, and put it together Store in a refrigerator at 4°C and protected from light.
[0027] 2. Staining: Use a micropipette to suck 15 μL of the staining solution obtained in step 1 onto the slide, and then pick the open flower that needs to be stained and observed from the inflorescence, use tweezers to pick 1 anther and put it directly on the slide In the staining solution, gently squeeze the anthers to disperse an appropriate amount of pollen, remove the broken anther wall and other debris, apply for a w...
Embodiment 2
[0030] Example 2: Fluorescence dyeing method of Chinese cabbage flower bud pollen
[0031] 1. Staining solution preparation: first dissolve Hoechst33258 fluorescent dye and sucrose in phosphate buffer saline (PBS, pH=7.4), prepare Hoechst33258 concentration 30μg / L, sucrose concentration 8wt% solution 1, and then prepare 70% ethanol ( v / v), ethyl acetic acid 10% (v / v), glycerol 20% (v / v) solution two, mix solution one and solution two in a volume ratio of 1:1 to obtain the staining solution, place Store in a refrigerator at 4°C and dark.
[0032] 2. Staining: Use a micropipette to suck 17μL of the staining solution prepared in step 1 onto the slide, then pick the flower buds to be stained and observed from the inflorescence, and use tweezers to peel 1 anther and put it directly on the slide In the staining solution, gently squeeze the anthers to disperse an appropriate amount of pollen, remove the broken anther wall and other debris, apply for a while (5-10 seconds) to mix the poll...
Embodiment 3
[0035] Example 3: Fluorescence dyeing method of flower bud pollen of Chinese cabbage
[0036] 1. Preparation of staining solution: first dissolve Hoechst33258 fluorescent dye and sucrose in phosphate buffer saline (PBS, pH=7.4) to prepare a solution of Hoechst33258 concentration of 40μg / L and sucrose concentration of 9wt%, and then prepare a solution containing 65% ethanol ( v / v), ethyl acetic acid 10% (v / v), glycerol 25% (v / v) solution two, mix solution one and solution two in a volume ratio of 1:1 to get the dyeing solution, and put it together Store in a refrigerator at 4°C and protected from light.
[0037] 2. Staining: Use a micropipette to suck 20μL of the staining solution obtained in step 1 onto the glass slide, and then pick the flower buds to be stained and observed from the inflorescence, and use tweezers to pick 2 anthers and put them directly on the glass slide In the staining solution, gently squeeze the anthers to disperse an appropriate amount of pollen, remove the...
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