Systems, devices and methods for determining the concentration of physiological fluid analytes are provided. The subject systems have a plurality of biosensor devices present on a disposable cartridge. Each biosensor device includes a biosensor and a skin penetration means. In practicing the subject methods, a movement means of the device is used to move each biosensor device in a first direction that provides for penetration of the skin-piercing means into a skin layer followed by movement of the biosensor in a second direction that provides for removal of the skin-piercing means from the skin layer, where this movement profile provides for physiological fluid access and analyte concentration determination by the analyte sensor means. The subject systems, devices and methods for using the same find use in determining the concentration of a variety of different physiological fluid analytes, and are particularly suited for use in detection of physiological fluid glucose concentration.