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Histology has been the gold standard for studying the cellular organization of body tissues and organs for several decades. The common procedures for histology studies generally include fresh tissue acquisition, tissue fixation, processing (for paraffin sectioning), embedding, sectioning, staining, and imaging. Various fixative methods, embedding materials, and sectioning methods have been developed to achieve different staining purpose. There are many histological staining methods used to study tissue characteristics and microscopic structures of tissue: including Immunohistochemistry (IHC) / Immunocytochemistry (ICC) / Immunofluorescence (IF) for detecting specific proteins, in situ hybridization (ISH) for specific DNA and RNA, Sudan staining for lipids, Sirius Red staining for collagens, routine Hematoxylin and Eosin (H&E) staining, and others. However, these techniques have some limitations: inefficient, two-dimensional, and low-resolution. Accordingly, scientists are putting efforts into developing more efficient, accurate, and high throughput histological techniques. Recently, several new histological techniques have been devised. These new techniques include the cytoplasm-specific / nucleus-specific X-ray staining for three-dimensional (3-D) histology of soft-tissue samples, the tissue clearing techniques (OPTIClear), and advanced in situ sequencing techniques: fluorescence in situ sequencing (FISSEQ), spatially resolved transcript amplicon read-out mapping (STARmap), and Slide-seq. These new techniques will significantly facilitate histological and histopathological research including identification of stem-cell inches, genome-wide spatial gene expression, tissue organization and functions. However, these new techniques also have some drawbacks: such as high background and generation of a plethora of data. Here, we summarize the most commonly used techniques and the latest advanced histological techniques, as well as their advantages and disadvantages.

the egyptian journal of histology

Histology Revolution: from inefficient, two-dimensional, and low-resolution techniques to high-throughput, three-dimensional and high-resolution techniques