Information processing apparatus and microscope system

Abstract

It is possible to more appropriately perform fluorescence separation. An information processing apparatus according to an embodiment includes: a fluorescence signal acquisition unit ( 112 ) that acquires a plurality of fluorescence spectra corresponding to each of a plurality of excitation lights having different wavelengths and irradiated to a fluorescence stained specimen ( 30 ), the fluorescence stained specimen ( 30 ) being created by staining a specimen ( 20 ) with a fluorescence reagent ( 10 ); a link unit ( 131 ) that generates a linked fluorescence spectrum by linking at least parts of the plurality of fluorescence spectra to each other in a wavelength direction; a separation unit ( 132 ) that separates the linked fluorescence spectrum into spectra for every fluorescent substance using a reference spectrum including a linked autofluorescence reference spectrum in which spectra of autofluorescent substances in the specimen are linked to each other in the wavelength direction and a linked fluorescence reference spectrum in which spectra of fluorescent substances in the fluorescence stained specimen are linked to each other in the wavelength direction; and an extraction unit ( 132 ) that updates the linked autofluorescence reference spectrum using the spectra for every fluorescent substance separated by the separation unit.

1 - 20 . (canceled) 
     
     
         21 . An information processing apparatus comprising:
 a fluorescence signal acquisition unit that acquires a plurality of fluorescence spectra corresponding to each of a plurality of excitation lights having different wavelengths and irradiated to a fluorescence stained specimen, the fluorescence stained specimen being created by staining a specimen with a fluorescence reagent;   a link unit that generates a linked fluorescence spectrum by linking at least parts of the plurality of fluorescence spectra to each other in a wavelength direction, wherein data segments of a predetermined spectral width are extracted from respective ones of the plurality of fluorescence spectra and the extracted data segments are linked together end-to-end in the wavelength direction to form the linked fluorescence spectrum;   a separation unit that separates the linked fluorescence spectrum into spectra for every fluorescent substance using a reference spectrum including a linked autofluorescence reference spectrum in which spectra of autofluorescent substances in the specimen are linked to each other in the wavelength direction and a linked fluorescence reference spectrum in which the spectra of the fluorescent substances in the fluorescence stained specimen are linked to each other in the wavelength direction; and   an extraction unit that updates the linked autofluorescence reference spectrum using the spectra for every fluorescent substance separated by the separation unit.   
     
     
         22 . The information processing apparatus according to  claim 21 , wherein
 the extraction unit extracts the linked autofluorescence reference spectrum from a linked autofluorescence spectrum generated by linking at least parts of a plurality of autofluorescence spectra to each other in the wavelength direction, the plurality of autofluorescence spectra being acquired by irradiating a section with the plurality of excitation lights, and the section being the same as or similar to the specimen.   
     
     
         23 . The information processing apparatus according to  claim 22 , wherein
 the extraction unit extracts the linked autofluorescence reference spectrum by performing non-negative matrix factorization using the linked autofluorescence spectrum generated by linking at least parts of a plurality of autofluorescence spectra to each other in the wavelength direction, the plurality of autofluorescence spectra being acquired by irradiating the section with the plurality of excitation lights, and the section being the same as or similar to the specimen.   
     
     
         24 . The information processing apparatus according to  claim 23 , wherein
 the extraction unit extracts the linked autofluorescence reference spectrum by setting an initial value in the non-negative matrix factorization using an autofluorescence spectrum acquired in advance.   
     
     
         25 . The information processing apparatus according to  claim 21 , wherein
 the separation unit separates the linked fluorescence spectrum into the spectra for every fluorescent substance using any one of a least square method or a weighted least square method using the reference spectrum.   
     
     
         26 . The information processing apparatus according to  claim 25 , wherein
 the separation unit separates the linked fluorescence spectrum into the spectra for every fluorescent substance by setting a matrix representing the linked fluorescence spectrum as Signal, setting a matrix representing the reference spectrum as St, setting a matrix representing a color mixture rate of each of the reference spectra in the linked fluorescence spectrum as a, and calculating the matrix a representing the color mixture rate when a sum of squares of values represented by the following Equation (1) becomes minimum:
   Signal− a*St   (1)
 
   
     
     
         27 . The information processing apparatus according to  claim 21 , wherein
 the separation unit separates the linked fluorescence spectrum into the spectra for every fluorescent substance using the reference spectrum including the linked autofluorescence reference spectrum and the linked fluorescence reference spectrum, which are calculated on a basis of the number of fluorescent molecules or the number of antibodies bound to the fluorescent molecules, or the reference spectrum including the linked autofluorescence reference spectrum and the linked fluorescence reference spectrum for each of the fluorescent molecules or for each of the antibodies.   
     
     
         28 . The information processing apparatus according to  claim 21 , wherein
 the separation unit separates the linked fluorescence spectrum into the spectra for every fluorescent substance by performing non-negative matrix factorization on the linked fluorescence spectra.   
     
     
         29 . The information processing apparatus according to  claim 28 , wherein
 the separation unit specifies a correspondence between the fluorescent substance and the extracted spectrum by calculating a product moment correlation coefficient with an initial value used for the non-negative matrix factorization for a spectrum extracted by the non-negative matrix factorization.   
     
     
         30 . The information processing apparatus according to  claim 21 , wherein
 the link unit corrects the plurality of fluorescence spectra, and links at least parts of the plurality of fluorescence spectra after being corrected to each other in the wavelength direction.   
     
     
         31 . The information processing apparatus according to  claim 30 , wherein
 the link unit corrects intensities of the plurality of fluorescence spectra.   
     
     
         32 . The information processing apparatus according to  claim 31 , wherein
 the link unit corrects the intensities of the plurality of fluorescence spectra by dividing the plurality of fluorescence spectra by an excitation power density.   
     
     
         33 . The information processing apparatus according to  claim 30 , wherein
 the link unit corrects a wavelength resolution of at least one of the plurality of fluorescence spectra to a wavelength resolution different from a wavelength resolution of another fluorescence spectrum.   
     
     
         34 . The information processing apparatus according to  claim 21 , wherein
 the link unit extracts fluorescence spectra in wavelength bands including intensity peaks from each of the plurality of fluorescence spectra, and generates the linked fluorescence spectrum by linking the extracted fluorescence spectra to each other.   
     
     
         35 . The information processing apparatus according to  claim 21 , wherein
 the linked fluorescence spectrum is discontinuously linked in the wavelength direction among the plurality of fluorescence spectra.   
     
     
         36 . The information processing apparatus according to  claim 21 , wherein
 the fluorescence signal acquisition unit acquires first image data which is obtained by imaging the fluorescence stained specimen and includes the plurality of fluorescence spectra, and   the separation unit separates the first image data into the spectra for every fluorescent substance by performing non-negative matrix factorization on a first Gram matrix of the first image data.   
     
     
         37 . The information processing apparatus according to  claim 36 , wherein
 the separation unit calculates the first Gram matrix by convolving a second Gram matrix of each of a plurality of second image data obtained by dividing the first image data.   
     
     
         38 . The information processing apparatus according to  claim 21 , wherein
 the fluorescence signal acquisition unit acquires first image data by imaging the specimen that is non-stained and irradiated with the excitation light, and   the extraction unit extracts the spectra for every autofluorescent substance from the first image data by performing non-negative matrix factorization on a first Gram matrix of the first image data, and updates the linked autofluorescence reference spectrum using the extracted spectra for every autofluorescent substance.   
     
     
         39 . The information processing apparatus according to  claim 21 , wherein wavelengths at boundaries between the linked together data segments are not continuous. 
     
     
         40 . A microscope system comprising: a light source that irradiates a fluorescence stained specimen with a plurality of excitation lights having different wavelengths, the fluorescence stained specimen being created by staining a specimen with a fluorescence reagent; an imaging apparatus that acquires a plurality of fluorescence spectra corresponding to each of the plurality of excitation lights; and a software used for processing using the plurality of fluorescence spectra, wherein
 the software is executed on an information processing apparatus, and   realizes:   generating a linked fluorescence spectrum by linking at least parts of the plurality of fluorescence spectra to each other in a wavelength direction, wherein data segments of a predetermined spectral width are extracted from respective ones of the plurality of fluorescence spectra and the extracted data segments are linked together end-to-end in the wavelength direction to form the linked fluorescence spectrum;   separating the linked fluorescence spectrum into spectra for every fluorescent substance using a reference spectrum including a linked autofluorescence reference spectrum in which spectra of autofluorescent substances in the specimen are linked to each other in the wavelength direction and a linked fluorescence reference spectrum in which the spectra of the fluorescent substances in the fluorescence stained specimen are linked to each other in the wavelength direction; and   updating the linked autofluorescence reference spectrum using the separated spectra for every fluorescent substance.