Porous silicon(PSi)was applied as a supporting substrate for stepwise covalent derivatization of undecylenic acid, N-hydroxysuccinimidyl ester(NHS-ester)and nitrilotriacetic acid(NTA).By taking the advantages of porous silicon as a supporting matrix such as high surface area to volume ratio,infrared transparency,porous semiconductors for laser desorption/ionization mass spectroscopy,and low fluorescence background,a multi-mode detection biochip prototype can be realized. We prepared such a protein microarray by spotting NTA microarray dots on NHS-ester derivatized PSi,converting the rest of chip area into poly(ethylene glycol)background,loading NiII,and finally affinity-binding histidine-tagged(His-tagged)proteins.With the multi-mode analyses of infrared spectroscopy,X-ray photoelectron spectroscopy(XPS),atomic force microscopy(AFM),matrix-assisted laser desorption/ionization mass spectroscopy(MALDI-MS),and fluorescence scanning,two example proteins,His-tagged thioredoxin-urodilatin and His-tagged aprotinin,were well qualified and quantified. Porous silicon (PSi) was applied as a supporting substrate for stepwise covalent derivatization of undecylenic acid, N-hydroxysuccinimidyl ester (NHS-ester) and nitrilotriacetic acid (NTA) .By taking the advantages of porous silicon as a supporting matrix such as high surface area to volume ratio, infrared transparency, porous semiconductors for laser desorption / ionization mass spectroscopy, and low fluorescence background, a multi-mode detection biochip prototype can be realized. We prepared such a protein microarray by spotting NTA microarray dots on NHS-ester derivatized PSi, converting the rest of chip area into poly (ethylene glycol) background, loading NiII, and finally affinity-binding histidine-tagged (His-tagged) proteins. Using the multi-mode analyzes of infrared spectroscopy, X-ray photoelectron spectroscopy XPS), atomic force microscopy (AFM), matrix-assisted laser desorption / ionization mass spectroscopy (MALDI-MS), and fluorescence scanning, two example proteins, His-tagged thioredoxin-urodilatin d His-tagged aprotinin, were well qualified and quantified.