我的英语不是很好有段英语翻译不出出来了希望哪位英语好的好心人帮我翻译一下吧，谢谢！！

Question

英语是：Immunohistochemistry Monolayered cultures of human fibroblasts on silanecoated
slides (Dako, Glostrup, Denmark) were fixed with 10% neutral buffered formalin
for 30 min. Acetone-fixed frozen slides were treated with blocking solution (Zymed,
San Francisco, CA, USA) for 30 min and incubated with monoclonal anti-α1(l)
procollagen aminoterminal extension peptide (SP1.D8) antibody (Developmental
Studies Hybridoma Bank, Iowa City, IA, USA) overnight in a humidified chamber at
4 °C. They were then incubated with biotinylated anti-mouse IgG antibody for 15 min
and incubated with horseradish-streptoavidin conjugate for 15 min. After washing in
PBS, color was developed using AEC (3-3-amino-9-ethylcarbazole (Zymed Laboratories,
San Francisco, CA, USA) for 5–10 min, and cell nuclei were counterstained with
Mayer's haematoxylin (Dako, Glostrup, Denmark).

Answer 1

首先tell sb to do sth, 否定形式就是 tell sb not to do sth
而你说为什么不能加don't。。。 don't read in bed 它的主语是he，而前面him 是宾格，不是主格，宾格是做宾语的
而please don't be late,kate 不能那样改，这是一个祈使句，只能用动词原形。只有动词接的是不定时，如tell sb to do, ask sb to do等，它变成否定只要在to do前面加上否定词