Biological substance detection method

Abstract

The present invention provides a biological substance detection method for specifically detecting a biological substance from a pathological specimen, by which method, when immunostaining using a fluorescent label and staining for morphological observation using a staining agent for morphological observation are simultaneously performed, the results of fluorescence observation and immunostaining can be assessed properly even if the fluorescent label and/or the staining agent is/are deteriorated by irradiation with an excitation light. The biological substance detection method according to the present invention is characterized in that the brightness retention rate of an immunostained part is in a range of 80% to 120% in relation to the brightness retention rate of apart stained for morphological observation when the fluorescent label used for the immunostaining is observed.

1 . A biological substance detection method for specifically detecting a biological substance from a pathological specimen, said method being characterized in that, in cases where immunostaining using a fluorescent label and staining for morphological observation using a staining agent for morphological observation are simultaneously performed, the brightness retention rate of an immunostained part is in a range of 80% to 120% in relation to the brightness retention rate of a part stained for morphological observation when said fluorescent label used for said immunostaining is observed. 
     
     
         2 . The biological substance detection method according to  claim 1 , wherein said fluorescent label is a fluorescent dye-containing nanoparticle whose maximum excitation wavelength is in a wavelength range of 550 to 620 nm and maximum emission wavelength is in a wavelength range of 580 to 700 nm. 
     
     
         3 . The biological substance detection method according to  claim 2 , wherein said fluorescent dye is a rhodamine-based dye molecule or an aromatic ring-based dye molecule. 
     
     
         4 . The biological substance detection method according to  claim 3 , wherein said fluorescent dye is perylene. 
     
     
         5 . The biological substance detection method according to  claim 2 , wherein a parent material of said fluorescent dye-containing nanoparticle is any of polystyrene, polyamide, polylactic acid, polyacrylonitrile, polyglycidyl methacrylate, polymelamine, polyurea, polybenzoguanamine, polyfuran, polyxylene, phenol resin, polysaccharide and silica. 
     
     
         6 . The biological substance detection method according to  claim 1 , wherein said staining agent has a maximum excitation wavelength in a wavelength range of 450 nm to 550 nm. 
     
     
         7 . The biological substance detection method according to  claim 6 , wherein said staining agent is eosin.