胡萝卜包埋制作拟南芥徒手切片

实验要点：

我亦无他，唯手熟尔！Practice makes perfect！

实验材料：

拟南芥花茎、胡萝卜、1号毛刷、吉列刀片、尖头镊子、美工刀、盖玻片、载玻片、200微升移液器、擦手纸、自来水

实验步骤：

1. 用美工刀将胡萝卜切成柱形，保持上表面水平。Cut the carrot into a column or prism with a knife. Keep the upper surface flat and horizontal. 

   

2. 用尖镊子在柱上戳一个小孔，小孔的直径近似于拟南芥花茎。Punch a hole on the prism with a sharp forcep. The diameter of the hole is similar to that of Arabidopsis stems.

   

3. 将花茎插入小孔。Insert one stem into the hole slightly with a hand or forcep.
   

   

   
   
4. 切除伸出在外面的多余花茎，确保胡萝卜小孔内有约0.5cm长的花茎段。Cut the stem that stretches out of the hole. Make sure the stem inside the hole about 0.5cm long.

   

5. 一只手握住胡萝卜，保持上表面水平，用吉列刀片水平切出一张薄片，切片尽量薄而不破。Hold the carrot with a hand and keep the upper surface horizontal. Cut a thin section with a razorblade (Gillette). Make the section as thin as possible but intact.
   

   

   
   
6. 切片会停留在刀片上，用半湿的1号毛刷轻轻将花茎切片挑出，置于含有200uL水的载玻片上。The section will be on the razorblade after cutting. Move the section with a half-wet painting brush slightly into 200uL water on the slide. 
   

   

   
   
7. 盖好盖玻片，在显微镜下观察切片。可以预先用间苯三酚对木质素染色（下图左），或直接在紫外光下观察木质素自发荧光（下图右）。Cover the cover glass. Examine the section under the microscope. Lignin is stained by phloroglucinol (left) or observed by self-fluorescence under UV light (right).