Method of analysing a sample for at least one analyte

A method of imaging at least one analyte in a biological sample for histology, cytology, or hybridizations used for support in diagnosing a subject, the method comprising:

• providing said sample being a section of tissue, a sample of cells, or a liquid obtained from said subject;
• contacting said sample with at least one targeting moiety or probe, wherein each different targeting moiety or probe of said at least one targeting moiety or probe specifically binds a different analyte of the at least one analyte, and wherein each different targeting moiety or probe of said at least one targeting moiety or probe is labelled with a different luminescent particle, wherein at least one of said luminescent particle is an upconverting particle;
• and
• obtaining a first image by detecting a signal from said luminescent particle associated with said at least one targeting moiety or probe bound to said sample;
• obtaining a second image from said sample using bright-field and wherein said second image is an image of said sample being counterstained or colored using a dye or a stain, wherein said dye or stain is not said upconverting particle;
• combining said first image with said second image to obtain a combined bright-field and co-registered fluorescent signal image comprising both a dye or stain under bright-field illumination and signals from said upconverting particle;
• wherein the first and second images are obtained using the same section of the sample to avoid issues with co-registration of images obtained from different sections of the sample, and
• wherein the luminescent particles are co-stained with the dye or stain used for bright-field.