System and method for tissue-wide single cell post-transcriptional profiling of multiple molecular targets

A method of mapping spatial distribution and expression of multiple molecular targets with individual cells within a two-dimensional tissue boundary, comprising:

• providing a molecular fishing system comprising an array of nanoprobes, wherein each nanoprobe is (i) functionalized with one or more molecular target fishing molecules for extracting the multiple molecular targets and (ii) crosslinked with one or more fluorescent dyes;
• contacting the array of nanoprobes with a surface of an acute tissue slice at an interface where the one or more molecular target fishing molecules and the one or more fluorescent dyes are in contact with a superficial layer of the acute tissue slice;
• extracting the molecular targets by the one or more molecular target fishing molecules from the acute tissue slice;
• exposing the other surface of the acute tissue slice to an imprint irradiation to imprint an outline of the acute tissue slice on the array;
• and removing the acute tissue slice from the array and subjecting the array to fluorescent imaging and subsequent barcoding,
• wherein the one or more fluorescent dyes have excitation and emission wavelengths in a range outside the imprint irradiation's spectrum and are cleavable from the nanoprobes by the imprint irradiation.