Due to significant advantages, the trend in the field of medical technology is moving towards minimally or even non-invasive examination methods. In this respect, optical methods offer inherent benefits, as does diffuse reflectance imaging (DRI). The present study attempts to prove the suitability of DRI—when implemented alongside a suitable setup and data evaluation algorithm—to derive information from anatomically correctly scaled human capillaries (diameter: $$10\,\upmu \hbox {m}$$
10
μ
m
, length: $$45\,\upmu \hbox {m}$$
45
μ
m
) by conducting extensive Monte–Carlo simulations and by verifying the findings through laboratory experiments. As a result, the method of shifted position-diffuse reflectance imaging (SP-DRI) is established by which average signal modulations of up to 5% could be generated with an illumination wavelength of $$\lambda =424\,\hbox {nm}$$
λ
=
424
nm
and a core diameter of the illumination fiber of $$50\,\upmu \hbox {m}$$
50
μ
m
. No reference image is needed for this technique. The present study reveals that the diffuse reflectance data in combination with the SP-DRI normalization are suitable to localize human capillaries within turbid media.