Hello

This protocol for Confocal microscopy used by Hong Tan et al  in their studies :

Collagen tube scaffolds containing AV explants were removed from the bioreactors and fixed in 4% paraformaldehyde at room temperature for 1 h. These tissues were embedded in 5% agarose gels and cross-sectioned into 300 μm thick sections using a vibratome (Oxford). Sections were then permeabilized with 0.25% triton X-100/PBS for 20 min at room temperature and blocked with 2% BSA/PBS overnight at 4 °C. Sections were stained for F-actin (phalloidin, # A12379, Invitrogen), tenascin C (#AB19013, Millipore), col1 (#AB752P, Millipore) and periostin (#AB1404.1, Abcam) using a concentration of 1:200 in BSA/PBS overnight at 4 °C. No primary antibody controls were incubated in PBS only overnight at 4 °C. The sections were then stained with Alexa-conjugated secondary antibodies (Invitrogen) and DAPI (#D21490, Invitrogen) overnight at concentrations of 1:200 and 1:5000, respectively, overnight at 4 °C. Sections were mounted with elevated cover slips in DABCO (#D2522, Sigma–Aldrich). Imaging was carried out using a Zeiss LSM 510 META confocal laser scanning microscope (Carl Zeiss). Images were collected using identical microscope settings for each channel. Gain was adjusted for each channel to prevent excessive pixel saturation; therefore, the reduced expression of tenascin C, col1, and periostin is not indicative of a lack of protein expression.

see full text in link . Hope help you 

Good Luck 

http://www.sciencedirect.com/science/article/pii/S0012160612006422?np=y