Rabbit anti human her2 primary antibody

The polyclonal rabbit anti-human HER2 primary antibody (Dako) was used for the tumour tissue biopsies, diluted 1:50 from the stock solution (10 μg ml⁻¹), and incubated at 4°C overnight. Goat anti-rabbit secondary antibody labeled with Alexa Fluor 680 (Invitrogen) was incubated for 1 h at room temperature at a protein concentration of 250 nM. Antibody conjugates were fixed in place with 2% paraformaldehyde. The slides were mounted with Fluoromount-G and covered with a coverslip for microscopy. Serial sections were stained with hematoxylin and eosin (H&E) using an autostainer (Leica). NIR fluorescence microscopy was performed on a Nikon TE300 with a 4-channel Nikon TE300. The excitation and emission filters used were 650/45 nm and 710/50 nm for Alexa 680 imaging, and 750/50 nm and 810/40 nm for NIR targeted tumour imaging, respectively.