组织切片对细胞核DNA含量检测的影响

目的探讨组织切片对图像分析仪测量细胞核DNA含量的影响。方法选取10只成年健康雄性小鼠,制备肝细胞涂片和肝组织切片,肝组织切片分成两部分,分别用于Feulgen染色和石蜡包埋,包埋后的组织采用垂直切片,图像分析仪测量切片实际厚度,依据切片机标识厚度和测量厚度分别分组,TIGER细胞图像分析仪分别测量肝细胞涂片和组织切片内肝细胞核的积分光密度(IOD)。结果肝细胞涂片内各DNA含量倍体肝细胞核IOD间的比值接近2和4,IOD之变异系数(CV值)<3.5,肝组织切片IOD间比值明显偏离2和4,IOD之CV值均>6;切片机标识厚度为4、6、8和10μm组织切片平均测量厚度分别为6.75、7.18、6.96和7.59μm,测量厚度最大值为9.25μm,最小值为4.62μm;依据切片机标识厚度分组中不同切片厚度相同DNA含量倍体肝细胞核的IOD值差异均无统计学意义;依据测量厚度重新分组后5、6微米组与7、8、9微米组IOD值间的差异具有统计学意义(P<0.05)。结论组织切片的实际厚度与切片机标识厚度间存在明显差异,本实验方法可较准确地判断组织切片厚度;厚组织切片测量结果优于薄组织切片,但与细胞涂片相比,厚组织切片仍难...     

不同器官石蜡包埋切片的切片压缩对比研究

目的比较研究不同器官石蜡包埋切片的切片压缩。方法从一成年雄性SD大鼠的肺、心、肝、睾丸和脊髓腰膨大各切取3个长、宽、高均约为2 mm的组织块,石蜡包埋后从每个组织块切取5～40μm厚(切片机设定厚度)的切片各3张进行苏木精染色,然后利用体视学图像系统的指针测微计测量每张切片染色后的实际厚度,以后一厚度(实际厚度)与前一厚度(切片设定厚度)之比作为切片压缩系数。结果与结论不同器官的平均压缩系数分别为84.4%(肺)、81.4%(心脏)、80.6%(肝脏)、77.9%(睾丸)和72.2%(脊髓),不同器官间有显著性差异。     

高温烤片对石蜡切片黏附、厚度与染色的影响

目的笔者实验室最近发现,高温烤片可有效解决甲基丙烯酸树脂包埋的组织切片的脱片问题,本文拟进一步研究确定高温烤片对石蜡包埋切片黏附、厚度及染色的影响。方法大鼠肾脏石蜡切片(切片机设定的切片厚度10μm)脱蜡后经90℃、140℃热板烤片30 min处理,然后用过碘酸-席夫试剂和苏木精染色,观察脱片情况、切片厚度及染色效果。大鼠脊髓石蜡切片(14μm厚)同样处理后分别进行小胶质细胞和突触素颗粒的免疫组化染色。结果与对照(未烤片)相比,90℃或140℃烤片都有效防止了肾脏石蜡切片从载玻片上脱落,但染色后的实际切片厚度减少了约50%(与设定切片厚度相比),染色变浅,结构清晰度欠佳(90℃烤片后),甚至有细胞消失(140℃烤片后);90℃烤片后免疫阳性小胶质细胞或突触素颗粒减少,非特异性染色加深,140℃烤片后几乎未见免疫阳性结构。结论高温烤片防脱片的方法可能不适用于石蜡切片。     

冷冻切片1 000例制作的体会

 目的 探讨冷冻切片在制作中的体会。方法 采用德国产Leica-CM1900型恒冷切片机：普通胶水、苏木精染色液、质量浓度为1 %醇溶性伊红、冰醋酸5 ml+质量浓度为10 %的中性甲醛10 ml+质量浓度为95 %的酒精85 ml混合固定液各100 ml盛于染色缸中备用。经常规冰冻切片取材、包埋、切片、染色后封片。结果 2003年至2005年共制作切片1 000例，诊断符合率达96.0 %以上。应用德国产Leica-CM1900型恒冷切片机制作冰冻切片，使用一次性日本羽毛切片刀，切片效果良好，切片完整，厚薄均匀无皱折，结构清晰，无空泡及空网状形成，亦无冰晶等现象。结论 冷冻切片质量好坏直接影响病理诊断，做好每一张切片都是一项关键性的技术操作。     

一种快速石蜡切片方法与快速冰冻切片方法的比较

我科在工作中一直用快速冷冻切片方法 ,但由于各种原因 ,影响着冰冻切片的质量 ,拖延病理报告发出的时间 ,也间接影响病人的预后。为了克服上述缺点 ,我们在实践中摸索出一种快速、简便、高质量的快速石蜡切片方法 ,现介绍如下。1　材料与方法1 1　材料　湖北产亚光牌ＹＧＴ -5Ｅ生物组织快速脱水仪 ,从固定到浸蜡的全过程都应加温至 80℃。1 2　方法　①将取材后的新鲜组织 (1ｃｍ× 0 .5ｃｍ× 0 .3ｃｍ)直接放入 10 %甲醛溶液中固定 3～ 5分钟 ,取出组织块后用刀修至厚度不超过 1 5ｍｍ。骨组织经 10 %甲醛溶液固定后加入10 %盐酸软化后…     

切片位置对兔睾丸组织体视学研究的影响

目的比较不同包埋介质内切片位置对成年新西兰大耳白兔睾丸组织体视学研究的影响。方法将成年雄性新西兰大耳白兔麻醉后,随机抽选一侧睾丸经Bouin液固定后从中央及两端（垂直于睾丸中心长轴）切取共三个平行组织薄片。将每个组织薄片平分为二,再随机将其中一块平分得到2个（1/4圆）组织块,分别随机采用羟乙基甲基丙烯酸树脂或石蜡包埋、切片和染色后,在低倍镜（10×）下分别对睾丸组织树脂切片（25μm）和石蜡切片（7μm）不同切片位置进行观察：紧贴白膜选择若干视野（所有视野间无任何重叠）,再向扇形切片中心连续选择3个视野,利用体视学方法估计各视野睾丸组织内生精小管的体积分数。结果石蜡切片较树脂切片估计的生精小管体积分数减少15.9%;树脂切片不同部位估计的生精小管体积分数无差异,而石蜡切片不同部位估计的结果存在差异：距离白膜愈远,体积分数愈小,近切片中心与近白膜相比显著减少了30.1%。结论石蜡包埋能引起睾丸组织不均匀皱缩,因此,当应用石蜡切片进行睾丸体积分数研究时,应考虑不均匀组织皱缩因素。     

提高淋巴结石蜡切片质量和染色效果的探讨

 目的　提高淋巴结制片质量。方法　用正丁醇代替乙醇和二甲苯脱水兼透明。将切片分别置于60 ％、70 %和80 ％正丁醇及95 ％乙醇各1 h,无水乙醇30 min,正丁醇Ⅰ2～3 h,正丁醇Ⅱ过夜。结果　与常规方法脱水制出的切片相比,该方法切片薄,细胞无重叠,细胞核核染色质形态清晰。结论　对于淋巴结组织制片,此方法优于常规染色方法,可以确保切片质量,从而避免由于切片质量问题导致的误诊     

厚、薄组织切片上细胞核体积测量结果的比较

目的: 研究组织切片厚度对细胞核体积测量结果的影响。方法: 收集鼻咽癌归档病例42例, 每个病例包括正常鼻咽上皮组织和3个癌巢或肿瘤区域, 制成4μm、8μm连续组织切片各一张。使用TIGER细胞图像分析仪分别测量4μm、8μm连续鼻咽癌组织切片上正常鼻咽上皮细胞核与肿瘤细胞核的体积。结果: 8μm组织切片上测得的细胞核体积明显大于4μm组织切片上测得的, 差异具有统计学意义(P<0 .05)。结论: 组织切片厚度影响细胞图像分析仪测量细胞核体积的结果; 采用图像分析仪测量细胞核内化学成分的含量应以完整细胞核为单位。     

冷冻切片制作的几点经验

目的 探讨理想的冷冻制片方法,以提高冷冻切片质量,从而提高病理诊断的准确性.方法 选取活体的组织标本,取材后直接冻结,经恒温式冷冻切片机切片,冷冻固定液固定,HE染色,剩余标本作石蜡切片对照.结果 冷冻切片质量好,染色鲜艳,镜下组织结构、细胞形态清晰,细胞核、质对比分明,质量与石蜡相同.结论 冷冻切片方法 简便、快速,制片质量较好,可以提高病理诊断的准确性.     

测量生物组织Epon薄切片厚度的研究

组织切片厚度的准确性对生物组织切片上的显微结构和化学成份定量分析结果的影响极大。至今，国内外已建立起的确定组织切片厚度的方法均有不足之处。本文将分光光度法的基本原理应用于组织切片厚度的测量，采用显微分光光度计和显微图像分析仪以及显微摄影测光表这些组织细胞形态和组织化学观察和定量分析中常用的仪器，对经锇酸固定的组织Ｅｐｏｎ薄切片的厚度进行测量。结果表明，这三种仪器均能用于测量Ｅｐｏｎ薄切片的厚度，而且经厚度测量的Ｅｐｏｎ薄切片尚能用于组织细胞形态和组织化学的定量分析；显微分光光度计和显微摄影测光表的测量灵敏度以及测量结果间没有显著差异，它们的测量结果比显微图像分析仪的测量结果更灵敏。本文讨论了这三种仪器在测量Ｅｐｏｎ薄切片厚度中的优缺点及其产生原因。     

深度学习在T1WI增强图像脑膜瘤分割中的价值

目的:探讨深度学习在T1WI增强图像脑膜瘤分割中的价值。方法:本研究共纳入326例经病理证实脑膜瘤患者，并将其分为训练集(n=194)、验证集(n=66)与测试集(n=66)。首先，对脑膜瘤进行手动勾画，再用图像和感兴趣区训练自动分割模型（基于SegResNet）。根据训练集、验证集平均dice loss值挑选最优模型，使用测试集评估模型表现并计算相关评价指标。结果:训练集、验证集、测试集的平均dice值分别为0.951、0.866、0.881。训练集、验证集、测试集的平均95% Hausdorff 距离（Hausdorff distance，95HD）分别为0.953、3.139、2.016。在测试集66个分割结果中，36个不需要修改，合格病灶比为54.5%。修改层数与肿瘤总层数分别为58、261，合格层数比为77.8%。结论:尽管自动分割模型并不能完全取代手动分割，但它在大部分层面分割表现优秀，可以有效减少不必要的工作。     

Automatic delineation of on-line head-and-neck computed tomography images: toward on-line adaptive radiotherapy

PURPOSE: To develop and validate a fully automatic region-of-interest (ROI) delineation method for on-line adaptive radiotherapy. METHODS AND MATERIALS: On-line adaptive radiotherapy requires a robust and automatic image segmentation method to delineate ROIs in on-line volumetric images. We have implemented an atlas-based image segmentation method to automatically delineate ROIs of head-and-neck helical computed tomography images. A total of 32 daily computed tomography images from 7 head-and-neck patients were delineated using this automatic image segmentation method. Manually drawn contours on the daily images were used as references in the evaluation of automatically delineated ROIs. Two methods were used in quantitative validation: (1) the dice similarity coefficient index, which indicates the overlapping ratio between the manually and automatically delineated ROIs; and (2) the distance transformation, which yields the distances between the manually and automatically delineated ROI surfaces. RESULTS: Automatic segmentation showed agreement with manual contouring. For most ROIs, the dice similarity coefficient indexes were approximately 0.8. Similarly, the distance transformation evaluation results showed that the distances between the manually and automatically delineated ROI surfaces were mostly within 3 mm. The distances between two surfaces had a mean of 1 mm and standard deviation of <2 mm in most ROIs. CONCLUSION: With atlas-based image segmentation, it is feasible to automatically delineate ROIs on the head-and-neck helical computed tomography images in on-line adaptive treatments.     

大块骨组织体视学研究的图像采集与分析

大块骨组织由于不脱钙切片制作技术难度高,应用体视学对大块骨组织整体形态进行图像定量分析的研究资料较少。随着生物材料、人工金属植入材料在骨科临床的大量应用,如何获取高质量的大块骨组织整体图像,并应用体视学原理进行较为精确的图像分析变成了迫切需要解决的问题。目的:寻求大块骨组织实用和有效的数字图像采集方法,使之应用体视学原理进行较为精确的图像分析。材料与方法:普通玻片上的大组织标本、硬组织切片机切取的不脱钙塑料包埋大块骨组织标本及旋转切割机切取的含金属植入物的塑料包埋磨片标本。分别采用普通显微镜、大视野显微镜、自动显微镜及图像拼接软件采集,同时采用胶片扫描仪、台式背透扫描仪及高分辩数码相机进行采集比较。结果:上述各方法在一定程度均能满足大块骨组织图像采集及体视学分析需求,同时在标本适应性、视野规模、成像质量方面总结出各自优、缺点以及相应技术方法。     

Evaluation of an atlas-based automatic segmentation software for the delineation of brain organs at risk in a radiation therapy clinical context.

BACKGROUND AND PURPOSE: Conformal radiation therapy techniques require the delineation of volumes of interest, a time-consuming and operator-dependent task. In this work, we aimed to evaluate the potential interest of an atlas-based automatic segmentation software (ABAS) of brain organs at risk (OAR), when used under our clinical conditions. MATERIALS AND METHODS: Automatic and manual segmentations of the eyes, optic nerves, optic chiasm, pituitary gland, brain stem and cerebellum of 11 patients on T1-weighted magnetic resonance, 3-mm thick slice images were compared using the Dice similarity coefficient (DSC). The sensitivity and specificity of the ABAS were also computed and analysed from a radiotherapy point of view by splitting the ROC (Receiver Operating Characteristic) space into four sub-regions. RESULTS: Automatic segmentation of OAR was achieved in 7-8 min. Excellent agreement was obtained between automatic and manual delineations for organs exceeding 7 cm3: the DSC was greater than 0.8. For smaller structures, the DSC was lower than 0.41. CONCLUSIONS: These tests demonstrated that this ABAS is a robust and reliable tool for automatic delineation of large structures under clinical conditions in our daily practice, even though the small structures must continue to be delineated manually by an expert.     

Comparison of intensity-modulated radiotherapy planning based on manual and automatically generated contours using deformable image registration in four-dimensional computed tomography of lung cancer patients

PURPOSE: To evaluate the implications of differences between contours drawn manually and contours generated automatically by deformable image registration for four-dimensional (4D) treatment planning. METHODS AND MATERIALS: In 12 lung cancer patients intensity-modulated radiotherapy (IMRT) planning was performed for both manual contours and automatically generated ("auto") contours in mid and peak expiration of 4D computed tomography scans, with the manual contours in peak inspiration serving as the reference for the displacement vector fields. Manual and auto plans were analyzed with respect to their coverage of the manual contours, which were assumed to represent the anatomically correct volumes. RESULTS: Auto contours were on average larger than manual contours by up to 9%. Objective scores, D(2%) and D(98%) of the planning target volume, homogeneity and conformity indices, and coverage of normal tissue structures (lungs, heart, esophagus, spinal cord) at defined dose levels were not significantly different between plans (p = 0.22-0.94). Differences were statistically insignificant for the generalized equivalent uniform dose of the planning target volume (p = 0.19-0.94) and normal tissue complication probabilities for lung and esophagus (p = 0.13-0.47). Dosimetric differences >2% or >1 Gy were more frequent in patients with auto/manual volume differences > or =10% (p = 0.04). CONCLUSIONS: The applied deformable image registration algorithm produces clinically plausible auto contours in the majority of structures. At this stage clinical supervision of the auto contouring process is required, and manual interventions may become necessary. Before routine use, further investigations are required, particularly to reduce imaging artifacts.     

人工智能技术在鼻咽癌放疗危及器官自动勾画中的应用研究

目的 评估基于人工智能技术的放疗危及器官自动勾画软件在临床应用中的价值.方法 选取2019年4月—2020年4月在自贡市第一人民医院肿瘤科治疗的30例鼻咽癌患者进行研究.首先,使用AccuContour软件勾画30例鼻咽癌患者的危及器官;然后使用瓦里安Eclipse v13.6计划系统人为手工勾画危及器官;最后将自动勾...     

41例甲状腺癌冰冻切片诊断分析

目的探讨甲状腺癌术中冰冻切片的诊断要点,提高术中冰冻切片的诊断水平。方法对我院206例甲状腺疾病中冰冻切片并经石蜡切片证实的41例甲状腺癌的病理资料进行回顾分析。结果41例甲状腺癌冰冻切片38例术中确诊,1例误诊为甲状腺不典型腺瘤,1例高度疑为乳头状癌而延迟诊断,1例因肿物钙化明显无法冰冻制片而漏诊。结论甲状腺癌术中冰冻切片诊断漏诊和过诊都可能导致严重的后果,术前了解病史,取材全面细致,掌握各型甲状腺癌的镜下特征,并由两位经验丰富的病理医师共同诊断是提高甲状腺癌冰冻切片病理诊断准确性的关键。     

Limited-data image registration for radiotherapy positioning and verification

PURPOSE: One benefit to having on-line CT imaging integrated into a radiotherapy system is that images can be collected with the patient in the treatment position. These images can then be automatically registered to planning images for improved positioning and verification. However, many such on-line imaging systems have a limited field of view (LFOV) that could potentially impair registration. Thus, the viability of automatic registration was investigated in the context of collecting on-line LFOV and also limited-slice CT images for radiotherapy. METHODS AND MATERIALS: Mutual information and two new voxel-based registration algorithms were tested to align LFOV and limited-slice prostate and breast images given known displacements. Success rates were tallied for different field-of-view sizes, slice distributions, and initial displacements. RESULTS: Most of the automatic registration algorithms tested were useful for solving these LFOV and limited-slice problems. Registration of LFOV images was generally successful, especially for fields of view of at least half the patient's size. For limited-slice images, success was more closely correlated to the slice spacing than to the number of slices used, with sparse slice spacing being preferable. CONCLUSIONS: Mutual information and other automatic registration algorithms have been identified as useful methods for registering LFOV and limited-slice radiotherapy images with planning CT images.     

Volumetric visualization of anatomy for treatment planning

PURPOSE: Delineation of volumes of interest for three-dimensional (3D) treatment planning is usually performed by contouring on two-dimensional sections. We explore the usage of segmentation-free volumetric rendering of the three-dimensional image data set for tumor and normal tissue visualization. METHODS AND MATERIALS: Standard treatment planning computed tomography (CT) studies, with typically 5 to 10 mm slice thickness, and spiral CT studies with 3 mm slice thickness were used. The data were visualized using locally developed volume-rendering software. Similar to the method of Drebin et al., CT voxels are automatically assigned an opacity and other visual properties (e.g., color) based on a probabilistic classification into tissue types. Using volumetric compositing, a projection into the opacity-weighted volume is produced. Depth cueing, perspective, and gradient-based shading are incorporated to achieve realistic images. Unlike surface-rendered displays, no hand segmentation is required to produce detailed renditions of skin, muscle, or bony anatomy. By suitable manipulation of the opacity map, tissue classes can be made transparent, revealing muscle, vessels, or bone, for example. Manually supervised tissue masking allows irrelevant tissues overlying tumors or other structures of interest to be removed. RESULTS: Very high-quality renditions are produced in from 5 s to 1 min on midrange computer workstations. In the pelvis, an anteroposterior (AP) volume rendered view from a typical planning CT scan clearly shows the skin and bony anatomy. A muscle opacity map permits clear visualization of the superficial thigh muscles, femoral veins, and arteries. Lymph nodes are seen in the femoral triangle. When overlying muscle and bone are cut away, the prostate, seminal vessels, bladder, and rectum are seen in 3D perspective. Similar results are obtained for thorax and for head and neck scans. CONCLUSION: Volumetric visualization of anatomy is useful in treatment planning, because 3D views can be generated without the need for segmentation. When relationships among anatomical structures, rather than geometric models of them, are important, volume rendering presents advantages. The presented algorithm is readily adaptable to distributed parallel implementation on a network of heterogeneous workstations.