The present invention relates to a liquid immersion micro-channel measurement device and measurement method which are based on trapezoidal incident structure prism incident-type silicon, and according to one embodiment of the present invention, the liquid immersion micro-channel measurement device based on trapezoidal incident structure prism incident-type silicon comprises: a micro-channel structure including a support and at least one micro-channel, which is formed on the support and has a sample detection layer to which a first bioadhesive material for detecting a first sample is fixed; a quadrangular pyramid-shaped prism formed on the upper part of the micro-channel structure; a sample injection unit for injecting, into the micro-channel, a buffer solution containing the first sample; a polarized light generation unit for emitting incident light polarized through the prism on the micro-channel at an incident angle that satisfies a p-wave non-reflection condition; and a polarized light detection unit for detecting, from the polarized incident light, a polarization change in a first refection light reflected from the sample detection layer, wherein the prism completely reflects, from the polarized incident light incident on the prism, on an upper boundary surface of the prism, second reflection light reflected from a lower boundary surface of the prism and a boundary surface of the buffer solution injected into the micro-channel.