They are loaded with an antigen by filling the inner compartment with a liquid vaccine formulation Cycloheximide followed by subsequent lyophilization. Vaccination with antigen filled bioneedles is performed by intramuscular or subcutaneous implantation under high velocity using compressed air. After implantation, the bioneedle dissolves, resulting in the release of the antigen. A clinical study showed that empty bioneedles are well tolerated by healthy volunteers during and after administration. No local toxicity other than tissue damage from bioneedle injection was observed at the site of implantation. Previous studies with tetanus toxoid and hepatitis B vaccines have shown that antigens delivered by bioneedles induce comparable or improved immune responses in mice compared to liquid vaccines delivered by conventional injection. Moreover, the lyophilized vaccine antigens in these bioneedles showed improved thermostability. This reduces the need for a cold-chain and allows long-term storage of vaccines. Furthermore, bioneedles are ideally suited for mass vaccinations. Vaccination with bioneedles is relatively easy, very quick and does not have the risk of needle stick injuries. Applicators will be low cost devices working on compressed air. Pressurizing the device can be done manually, which make it ideal for use in developing countries. Furthermore, cost assessments have indicated that bioneedle applicator devices could be supplied free of charge for the use in public health care in developing countries. In this current study, we compared the immunogenicity of influenza vaccine filled bioneedles with the immunogenicity of conventional liquid influenza vaccines in mice. In order to identify the most potent influenza vaccine formulation for inclusion in bioneedles, we included four types of non-adjuvanted influenza vaccine. Furthermore, the thermostability of the lyophilized influenza vaccine formulations was evaluated. This study demonstrates that influenza vaccines delivered by bioneedles elicit equal or improved immune responses in C57BL/ 6 mice compared to conventional liquid vaccines. Furthermore, lyophilization of the different influenza vaccine types formulated with trehalose and HBS greatly improved the heat stability of the influenza vaccines. The four influenza vaccine types were selected for their differences in compositions and particulate organization. All vaccines were produced from a single virus batch, enabling a fair comparison between the vaccines. Comparative immunogenicity studies remain few, and studies on subcutaneous influenza vaccines are mostly limited to a single vaccine type. This knowledge gap makes it difficult to preselect the most suitable influenza vaccine candidate for delivery with bioneedles. Therefore, a head-to-head comparison between these four vaccines was performed in this study. Characterization of the produced influenza vaccines confirmed that WIV and split vaccines contained all viral components, whereas virosome and subunit vaccines only contained the membrane proteins HA and NA.