Dear Kuo-Kuang Wen,
For the primary or secondary antibody you can prepare 300 microliters (of diluted antibody at the working concentration) for each slide;
you can drop this solution at the center of the slide and gently put on top of it a glass coverslip, avoiding bubble formation, in order to facilitate the antibody solution spreading over the entire surface of the bottom slide.
Following incubation remove gently the coverslip and proceed with washing and further steps.
Bests
Luigi Alfano
2020-12-09 06:53:35