A method for obtaining a calibration curve for quantifying protein copy number in fluorescence-based super resolution microscopy which comprises a) incubating a DNA origami immobilized on a support, wherein the DNA origami comprises handle oligonucleotides protruding from said DNA origami, said handle oligonucleotides being attached to the DNA origami at predetermined positions and at least one tag, with a protein of interest functionalized with oligonucleotides complementary to the handles protruding from said DNA origami, in conditions allowing the hybridization between the oligonucleotides attached to the DNA origami and the oligonucleotides attached to the protein of interest, b) recording a super resolution image of the protein of interest which colocalizes with the tag of the DNA origami, c) clustering the image obtained in step b) and identifying the clusters separated by the distance between the handles to obtain the number clusters in said image obtained in step b), d) fitting a generic probability distribution function depending on a set of parameters μ to the distribution of the number of localizations x for one predetermined cluster, f 1 μ ; x and extending it iteratively to larger clusters by using the equation for n= 2, 3... Nmax f n = f n − 1 ⊗ f 1 where ⊗ represent the convolution in respect to the x variables, between two functions and n max is a predetermined maximum number of clusters, and e) obtaining a calibration curve by the parameters determined through the fitting procedure described in d), wherein steps d) and e) are executed by a computer, and wherein the super resolution image of the protein of interest is obtained by detecting said protein with a fluorescent antibody, fluorescent nanobody or fluorescent halo/snap tag specific for the protein of interest. a) incubating a DNA origami immobilized on a support, wherein the DNA origami comprises handle oligonucleotides protruding from said DNA origami, said handle oligonucleotides being attached to the DNA origami at predetermined positions and at least one tag, with a protein of interest functionalized with oligonucleotides complementary to the handles protruding from said DNA origami, in conditions allowing the hybridization between the oligonucleotides attached to the DNA origami and the oligonucleotides attached to the protein of interest, b) recording a super resolution image of the protein of interest which colocalizes with the tag of the DNA origami, c) clustering the image obtained in step b) and identifying the clusters separated by the distance between the handles to obtain the number clusters in said image obtained in step b), d) fitting a generic probability distribution function depending on a set of parameters μ to the distribution of the number of localizations x for one predetermined cluster, f 1 μ ; x and extending it iteratively to larger clusters by using the equation for n= 2, 3... Nmax f n = f n − 1 ⊗ f 1 where ⊗ represent the convolution in respect to the x variables, between two functions and n max is a predetermined maximum number of clusters, and e) obtaining a calibration curve by the parameters determined through the fitting procedure described in d), wherein steps d) and e) are executed by a computer, and wherein the super resolution image of the protein of interest is obtained by detecting said protein with a fluorescent antibody, fluorescent nanobody or fluorescent halo/snap tag specific for the protein of interest. <EOS>