Multiphoton microscopic imaging of adipose tissue based on second‐harmonic generation and two‐photon excited fluorescence

Abstract

The fresh adipose tissue was investigated by the use of multiphoton microscopy (MPM) based on two-photon excited fluorescence and second-harmonic generation (SHG). Microstructure of collagen and adipose cells in the adipose tissue is clearly imaged at a subcellular level with the excitation light wavelengths of 850 and 730 nm, respectively. The emission spectrum of collagen SHG signal and NADH and FAD fluorescence signal can also be obtained, which can be used to quantify the content of collagen and adipose cells and reflect the degree of pathological changes when comparing normal tissue with abnormal adipose tissue in the same condition. The results indicate that MPM has the potential to be applied to investigate the adipose tissue and can be used in the research field of lipid and connective tissues.

Fractal Analysis of Two‐Photon Microscopic Images for Diagnosis of Nasopharyngeal Cancer

“…Two‐photon microscopy (TPM) imaging is currently a powerful and well‐established tool, as it provides rapid, real‐time, and noninvasive “optical biopsy” evaluation of tissue. Our previous studies have demonstrated that TPM technique can not only enable direct noninvasive visualization of the distribution of extracellular matrix components, but monitor the morphological and functional changes of cells, and good agreement was obtained between TPM image and the corresponding H&E staining image (Huang et al ., ; Chen et al ., ). Even so, for more objective and reliable pathologic diagnosis, it is essential to quantify morphological changes based on the TPM images.…”

Heterogeneity in Subcutaneous Adipose Tissue Morphology and Metabolic Complications in Overweight and Obese Women

“…One previous study demonstrated the feasibility for visualization of adipocytes and collagen in adipose tissue by MPAM/SHGM. 13 That study showed adipocytes and collagen could be identified by MPAM and SHGM, respectively, but did not indicate the site of AT origin or status of obesity. Given the complex 3D architecture and makeup of AT and the potential role that the extracellular matrix may play in remodeling and function of this tissue, 14 methodologies that provide indication of spatial organization between AT constituents could be highly beneficial to the characterization of intact AT but also be used to better understand the changes associated with obesity and insulin resistance.…”

Two-photon excited fluorescence imaging of the pancreatic solid pseudopapillary tumor without hematoxylin and eosin stains

“…Recently, two‐photon excited fluorescence (TPEF) microscopy has been applied to image tissue architecture and cellular morphology of unstained tissue samples in biomedical research (Chen et al ., ). This technique presents a non‐invasive and high‐resolution laser scanning enabling visualization of cellular and subcellular structures without any exogenous fluorescent agents while maintaining tissue viability (Zipfel et al ., ; Wilder et al ., ; Huang et al ., ). Compared with traditional optical imaging, TPEF microscopy has the advantages of deeper penetration depth, higher resolution, lower photo‐bleaching, and photo‐toxicity.…”