Multiphoton microscopic imaging of histological sections without hematoxylin and eosin staining differentiates carcinoma in situ lesion from normal oesophagus

Multiphoton microscopy (MPM) has become a powerful, important tool for tissues imaging at the molecular level. In this paper, this technique was extended to histological investigations, differentiating carcinoma in situ (CIS) lesion from normal oesophagus by imaging histological sections without hematoxylin and eosin (H&E) staining. The results show that the histology procedures of dehydration, paraffin embedding, and de-paraffinizing highlighted two photon excited fluorescence of cytoplasm and nucleolus of epithelial cell and collagen in stroma. MPM has the ability to identify the characteristics of CIS lesion including changes of squamous cells and full epithelium, identification of basement membrane, especially prominent nucleolus. The studies described here show that MPM has the potential for future retrospective studies of tumor staging by employing on histological section specimens without H&E staining.

[1]  Melissa C Skala,et al.  Multiphoton microscopy of endogenous fluorescence differentiates normal, precancerous, and cancerous squamous epithelial tissues. , 2005, Cancer research.

[2]  Dong Li,et al.  Multimodal nonlinear optical microscopy improves the accuracy of early diagnosis of squamous intraepithelial neoplasia , 2013, Journal of biomedical optics.

[3]  Jun Nagata,et al.  Multiphoton imaging can be used for microscopic examination of intact human gastrointestinal mucosa ex vivo. , 2008, Clinical gastroenterology and hepatology : the official clinical practice journal of the American Gastroenterological Association.

[4]  A. Jemal,et al.  Cancer statistics, 2012 , 2012, CA: a cancer journal for clinicians.

[5]  Shuangmu Zhuo,et al.  Multimode nonlinear optical imaging of the dermis in ex vivo human skin based on the combination of multichannel mode and Lambda mode. , 2006, Optics express.

[6]  Shuangmu Zhuo,et al.  Extracting diagnostic stromal organization features based on intrinsic two-photon excited fluorescence and second-harmonic generation signals. , 2009, Journal of biomedical optics.

[7]  Chen-Yuan Dong,et al.  Evaluating cutaneous photoaging by use of multiphoton fluorescence and second-harmonic generation microscopy. , 2005, Optics letters.

[8]  P R Taylor,et al.  Black oesophagus—cause? , 2005, Gut.

[9]  Shuangmu Zhuo,et al.  Establishing diagnostic features for identifying the mucosa and submucosa of normal and cancerous gastric tissues by multiphoton microscopy. , 2011, Gastrointestinal endoscopy.

[10]  Minsue T. Chen,et al.  Hematoxylin and Eosin Tissue Stain in Mohs Micrographic Surgery: A Review , 2011, Dermatologic surgery : official publication for American Society for Dermatologic Surgery [et al.].

[11]  Virginijus Barzda,et al.  Nonlinear multicontrast microscopy of hematoxylin-and-eosin-stained histological sections. , 2010, Journal of biomedical optics.

[12]  Shuangmu Zhuo,et al.  Depth-cumulated epithelial redox ratio and stromal collagen quantity as quantitative intrinsic indicators for differentiating normal, inflammatory, and dysplastic epithelial tissues , 2010 .

[13]  Shuangmu Zhuo,et al.  Label-free multiphoton imaging and photoablation of preinvasive cancer cells , 2012 .

[14]  J. Kiškis,et al.  IMAGING OF HISTOLOGICAL SECTIONS OF THE HUMAN INTESTINE BY MULTIMODAL CARS MICROSCOPE , 2012 .

[15]  Paolo P. Provenzano,et al.  Fluorescence Lifetime Imaging of Endogenous Fluorophores in Histopathology Sections Reveals Differences Between Normal and Tumor Epithelium in Carcinoma In Situ of the Breast , 2009, Cell Biochemistry and Biophysics.

[16]  Chen-Yuan Dong,et al.  Discrimination of basal cell carcinoma from normal dermal stroma by quantitative multiphoton imaging. , 2006, Optics letters.

[17]  W. Webb,et al.  Nonlinear magic: multiphoton microscopy in the biosciences , 2003, Nature Biotechnology.

[18]  P. Taylor,et al.  Squamous esophageal histology and subsequent risk of squamous cell carcinoma of the esophagus.A prospective follow‐up study from Linxian, China , 1994, Cancer.

[19]  Guannan Chen,et al.  Detecting the imaging characteristics of colorectal carcinoma invading the muscularis propria with multiphoton microscopy , 2012 .