Rapid virtual hematoxylin and eosin histology of breast tissue specimens using a compact fluorescence nonlinear microscope.
Rapid virtual hematoxylin and eosin histology of breast tissue specimens using a compact fluorescence nonlinear microscope.
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DOI：
10.1038/labinvest.2017.116
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发表时间：
2018-01
期刊：
影响因子：
--
通讯作者：
Fujimoto JG
中科院分区：
文献类型：
--
作者：
Cahill LC;Giacomelli MG;Yoshitake T;Vardeh H;Faulkner-Jones BE;Connolly JL;Sun CK;Fujimoto JG
在接受乳腺癌保乳手术的患者中，高达40%的患者由于切缘接近或为正，需要再次手术。标准石蜡包埋组织学评估手术切缘所需的漫长过程使其无法在手术中使用，因此，快速评估手术病理的技术可以通过减少所需的手术次数来改进乳腺癌的治疗。我们演示了乳腺癌手术标本的实时组织学评估，方法是用类似于苏木精伊红(H&E)的丫啶橙(AO)和磺胺类霍丹明101(SR101)对标本进行染色，然后使用紧凑的飞秒光纤激光器用荧光非线性显微镜(NLM)对标本进行成像。使用视频率计算光吸收模型来实时和三维地生成逼真的组织虚拟H&E图像。NLM成像可以在组织表面以下100微米的深度进行，这一点很重要，因为许多外科标本需要进行亚表面评估，因为组织表面存在电灼、外科墨水或标本处理碎片造成的伪影。我们通过与福尔马林固定、石蜡包埋(FFPE)组织学相比较的NLM图像的专家评审来验证该方法。正常的末梢导管小叶单位、纤维和脂肪间质实质、炎症、浸润性癌、原位小叶和导管癌等诊断上的重要征象在NLM图像中与标准FFPE H&E组织学确定的病理相关。我们证明，在FFPE处理后，AO和SR101被提取到无法检测的水平，并且荧光原位杂交(FISH)HER2扩增状态不受NLM成像程序的影响。这种方法有可能使手术切除的实时组织学指导更具成本效益。
Up to 40% of patients undergoing breast conserving surgery for breast cancer require repeat surgeries due to close to or positive margins. The lengthy processing required for evaluating surgical margins by standard paraffin embedded histology precludes its use during surgery and therefore, technologies for rapid evaluation of surgical pathology could improve the treatment of breast cancer by reducing the number of surgeries required. We demonstrate real-time histological evaluation of breast cancer surgical specimens by staining specimens with acridine orange (AO) and sulforhodamine 101 (SR101) analogously to hematoxylin and eosin (H&E) and then imaging the specimens with fluorescence nonlinear microscopy (NLM) using a compact femtosecond fiber laser. A video-rate computational light absorption model was used to produce realistic virtual H&E images of tissue in real time and in three dimensions. NLM imaging could be performed to depths of 100 µm below the tissue surface, which is important since many surgical specimens require subsurface evaluation due to artifacts on the tissue surface from electrocautery, surgical ink or debris from specimen handling. We validate this method by expert review of NLM images compared to formalin fixed, paraffin embedded (FFPE) H&E histology. Diagnostically important features such as normal terminal ductal lobular units, fibrous and adipose stromal parenchyma, inflammation, invasive carcinoma, and in-situ lobular and ductal carcinoma were present in NLM images associated with pathologies identified on standard FFPE H&E histology. We demonstrate that AO and SR101 were extracted to undetectable levels after FFPE processing and fluorescence in situ hybridization (FISH) HER2 amplification status was unaffected by the NLM imaging protocol. This method potentially enables cost-effective, real-time histological guidance of surgical resections.
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影响因子：
3.7
作者：
Giacomelli MG;Husvogt L;Vardeh H;Faulkner-Jones BE;Hornegger J;Connolly JL;Fujimoto JG
通讯作者：
Fujimoto JG
影响因子：
1.9
作者：
Millard, AC;Wiseman, PW;Müller, M
通讯作者：
Müller, M
影响因子：
2.4
作者：
Chung, VQ;Dwyer, PJ;Jiang, SB
通讯作者：
Jiang, SB
影响因子：
3.8
作者：
Kobat, Demirhan;Durst, Michael E.;Xu, Chris
通讯作者：
Xu, Chris
影响因子：
5.6
作者：
FERREIRO, JA;GISVOLD, JJ;BOSTWICK, DG
通讯作者：
BOSTWICK, DG