In life sciences, the problem of very small volume of sample, analytes, and reagents is often faced. Micro-fluidic technology is ideal for handling costly and difficult-to-obtain samples, analytes, and reagents, because it requires very small volume of samples, in order of microL or even nL. Among many types of optical techniques commonly used for biosensing in microfluidic chip, fluorescence detection technique is the most common. The standard free-space detection techniques used to detect fluorescence emission from microfluidic channel often suffer issues like scattering noise, crosstalks, misalignment, autofluorescence of substrate, and low collection efficiency. This chapter describes two fluorescence detection methods, based on in-fiber microchannels and in-fiber grooves, which can solve those problems, as the techniques integrate the excitation and emission light paths, and the sensing part. Utilizing an optical fiber as a sensing component makes these detection techniques suitable for lab-on-a-chip or microTAS applications.