The clinical management of amyloidosis is based on the treatment of the underlying etiology, and accurate identification of the protein causing the amyloidosis is of paramount importance. Current methods used for typing of amyloidosis such as immunohistochemistry have low specificity and sensitivity. In this study, we report development of a highly specific and sensitive novel test for typing of amyloidosis in routine clinical biopsy specimens. Our approach combines specific sampling by laser microdissection (LMD) and analytical power of tandem mass spectrometry (MS) based proteomic analysis. We studied 50 cases of amyloidosis which were well-characterized by gold standard clinicopathological criteria (training set) and, an independent validation set comprising 41 cases of cardiac amyloidosis. LMD/MS identified the amyloid type with 100% specificity and sensitivity in the training set and was conclusive in 98% of the validation set. LMD/MS method will enhance our ability to type amyloidosis accurately in clinical biopsy specimens.