Production of tissue microarrays, immunohistochemistry staining and digitalization within the human protein atlas.
The  tissue microarray (TMA) technology provides the means for high-throughput  analysis of multiple tissues and cells. The technique is used within the Human  Protein Atlas project for global analysis of protein expression patterns in  normal human tissues, cancer and cell lines. Here we present the assembly of 1  mm cores, retrieved from microscopically selected representative tissues, into  a single recipient TMA block. The number and size of cores in a TMA block can  be varied from approximately forty 2 mm cores to hundreds of 0.6 mm cores. The  advantage of using TMA technology is that large amount of data can rapidly be  obtained using a single immunostaining protocol to avoid experimental  variability. Importantly, only limited amount of scarce tissue is needed, which  allows for the analysis of large patient cohorts (1 2). Approximately 250 consecutive  sections (4 μm  thick) can be cut from a TMA block and used for immunohistochemical staining to  determine specific protein expression patterns for 250 different antibodies. In  the Human Protein Atlas project, antibodies are generated towards all human  proteins and used to acquire corresponding protein profiles in both normal  human tissues from 144 individuals and cancer tissues from 216 different  patients, representing the 20 most common forms of human cancer. Immunohistochemically  stained TMA sections on glass slides are scanned to create high-resolution  images from which pathologists can interpret and annotate the outcome of  immunohistochemistry. Images together with corresponding pathology-based  annotation data are made publically available for the research community  through the Human Protein Atlas portal (www.proteinatlas.org) (Figure 1) (3 4). The Human Protein Atlas  provides a map showing the distribution and relative abundance of proteins in  the human body. The current version contains over 11 million images with  protein expression data for 12.238 unique proteins, corresponding to more than 61%  of all proteins encoded by the human genome.
Department of Immunology, Genetics and Pathology, Science for Life Laboratory, Uppsala University, Sweden.
Caroline
C
Kampf
Ingmarie
I
Olsson
Urban
U
Ryberg
Evelina
E
Sjöstedt
Fredrik
F
Pontén
D030562:Databases, Protein; D006801:Humans; D007150:Immunohistochemistry / Q000379:methods; D009363:Neoplasm Proteins / Q000032:analysis; D009369:Neoplasms / Q000737:chemistry; D016612:Paraffin Embedding; D011506:Proteins / Q000032:analysis*; D040901:Proteomics / Q000379:methods; D013194:Staining and Labeling / Q000379:methods*; D046888:Tissue Array Analysis / Q000295:instrumentation / Q000379:methods*
D009363:Neoplasm Proteins; D011506:Proteins