Comparison of Hematoxylin-eosin Staining and Methyl Violet Staining for Displaying Ghost Cells

Purpose:.To compare the merits and limitations of hematoxylin-eosin(HE) and methyl violet staining for displaying ghost cells from vitreous or aqueous humor.Methods:.A specimen containing ghost cells was adjusted to five different concentrations:(12×104,.10×104,.8×104, 6×104and 4×104cells / ml) and subjected to smearing and methyl violet and HE staining..The staining results were observed by light microscopy.Results: The ghost cells were readily observed at a cell density of > 8×104cells / ml with methyl violet staining,.but only a few cells were occasionally seen at lower cell densities..In contrast,.ghost cells were seen at all cell densities with HE staining.Conclusion: Methyl violet staining is more rapid and simpler for the identification of ghost cells, but its staining color more readily fades, the slides cannot be stored, and it is only effective at a cell density of > 8 ×104cells / ml. In contrast,.HE staining is more time-consuming but it can display cell morphology and distinguish cell components more explicitly and slides can be permanently stored. HE staining has advantages over methyl violet staining in detecting the ghost cells when the concentration is < 8×104cells / ml.

Hepatic and renal effects of oral stingless bee honey in a streptozotocin-induced diabetic rat model

BACKGROUND Diabetes is known damage the liver and kidney,leading to hepatic dysfunction and kidney failure.Honey is believed to help in lowering the blood glucose levels of diabetic patients and reducing diabetic complications.However,the effect of stingless bee honey(SBH)administration in relieving liver and kidney damage in diabetes has not been well-studied.AIM To investigate the effect of SBH administration on the kidney and liver of streptozotocin-induced(STZ;55 mg/kg)diabetic Sprague Dawley rats.METHODS The rats were grouped as follows(n=6 per group):non-diabetic(ND),untreated diabetic(UNT),metformin-treated(MET),and SBH+metformin-treated(SBME)groups.After successful diabetic induction,ND and UNT rats were given normal saline,whereas the treatment groups received SBH(2.0 g/kg and/or metformin(250 mg/kg)for 12 d.Serum biochemical parameters and histological changes using hematoxylin and eosin(H&E)and periodic acid–Schiff(PAS)staining were evaluated.RESULTS On H&E and PAS staining,the ND group showed normal architecture and cellularity of Bowman’s capsule and tubules,whereas the UNT and MET groups had an increased glomerular cellularity and thickened basement membrane.The SBH-treated group showed a decrease in hydropic changes and mild cellularity of the glomerulus vs the ND group based on H&E staining,but the two were similar on PAS staining.Likewise,the SBME-treated group had an increase in cellularity of the glomerulus on H&E staining,but it was comparable to the SBH and ND groups on PAS staining.UNT diabetic rats had tubular hydropic tubules,which were smaller than other groups.Reduced fatty vacuole formation and dilated blood sinusoids in liver tissue were seen in the SBH group.Conversely,the UNT group had high glucose levels,which subsequently increased MDA levels,ultimately leading to liver damage.SBH treatment reduced this damage,as evidenced by having the lowest fasting glucose,serum alanine transaminase,aspartate transaminase,and alkaline phosphatase levels compared to other groups,although the levels of liver enzymes were not statistically significant.CONCLUSION The cellularity of the Bowman’s capsule,as well as histological alteration of kidney tubules,glomerular membranes,and liver tissues in diabetic rats after oral SBH resembled those of ND rats.Therefore,SBH exhibited a protective hepatorenal effect in a diabetic rat model.

DNA extraction from archived hematoxylin and eosin-stained tissue slides for downstream molecular analysis

BACKGROUND Histopathologically stained archived tissue slides are stored in hospital archives for years to decades.They are the largest available source of biological materials and are a potentially useful resource that can be used for retrospective epidemiological studies.DNA recovered from the slides can be used for several downstream molecular processes including polymerase chain reaction,single nucleotide polymorphism analysis,and whole genome sequencing.The DNA from these slides can be utilized to compare gene signatures of normal and diseased tissues.However,extraction of high-quality DNA from archived stained hematoxylin and eosin(H&E)slides remains challenging.AIM To standardize a new protocol for extracting DNA from archived H&E-stained tissue slides for further molecular assays.METHODS A total of 100 archived H&E-stained cancer slides were subjected to a total of five methods of DNA extraction.Methods were varied in the deparaffinization step,tissue rehydration,duration of lysis,and presence or absence of proteinase K.The extracted DNA was quantified using a NanoDrop spectrophometer and the quality was analyzed by agarose gel electrophoresis.Then each sample was subjected to polymerase chain reaction(PCR)to amplify the internal control gene GAPDH,thereby confirming the DNA intactness,which could be further utilized for other downstream applications.RESULTS Of the five different methods tested,the third method wherein xylene was used for tissue deparaffinization followed by 72 h of digestion and without proteinase K inactivation yielded the highest amount of DNA with good purity.The yield was significantly higher when compared to other methods.In addition,90%of the extracted DNA showed amplifiable GAPDH gene.CONCLUSION Here we present a step-by-step,cost-effective,and reproducible protocol for the extraction of PCR-friendly DNA from archived H&E-stained cancer tissue slides that can be used for further downstream molecular applications.

二次脱钙法在临床病理骨组织HE制片中的应用研究

目的 探讨二次脱钙法在临床病理骨组织苏木精-伊红(HE)制片中的应用价值。方法 收集2022年6-12月该院病理科脱钙不充分、骨组织块或含钙化组织的蜡块100块,制成病理切片,根据脱钙方法的不同将病理切片分为对照组和研究组,对照组采用常规脱钙法,研究组采用二次脱钙法,分析2种方法制片效果的差异。结果 研究组病理切片HE制片结构完整性、HE染色满意度、切片优良率均明显高于对照组,差异均有统计学意义(P<0.05)。结论 二次脱钙法在临床病理骨组织HE制片中具有较高的应用价值。

Digitally reinforced hematoxylin-eosin polarization technique in diagnosis of rectal amyloidosis

AIM:To investigate the efficacy of the digitally reinforced hematoxylin-eosin polarization(DRHEP)technique for detection of amyloidosis in rectal biopsies.METHODS:One hundred hematoxylin-eosin(HE)stained rectal biopsies with Congo-red(CR)-positive amyloid depositions and 50 control cases with CRnegative amyloid-mimicking areas were scanned blinded to the CR results for amyloid depositions under both bright and polarized light,and digitally photographed using the DRHEP technique,to accentuate the faint birefringence observed in HE slides under polarization.The results of DRHEP and HE evaluation were statistically correlated with CR polarization results with respect to presence and localization of amyloid deposits as well as amyloid types.RESULTS:Amyloid deposits showed yellowish-green birefringence by DRHEP,which allowed identification of amyloidosis in 41 HE-unsuspected cases(P=0.016),31 of which only had vascular deposits.True positivity was higher,and false negativity and positivity were lower by DRHEP,compared to evaluation by HE(69%,31%,and 0.8%vs 33%,67%,and 33%,respectively;P<0.0001).The sensitivity,specificity,accuracy,and positive and negative predictive values for DRHEP were69%,98%,78.6%,98.5%,and 61.25%,respectively.Reasons for DRHEP false negativity were presence of extensive background birefringence in 12 cases,absence of CR birefringent vessel in 3 cases,and missing of the tiny deposits in 9 cases,which could be improved by experience,especially in the latter case.No correlation was found between age,gender,sites of deposits,or amyloid types.CONCLUSION:The DRHEP technique improves diagnostic accuracy when used as an adjunct or a prior step to CR staining,especially for cases with limited tissues for further analysis.

Diagnostic value of endothelial markers and HHV-8 staining in gastrointestinal Kaposi sarcoma and its difference in endoscopic tumor staging

AIM: To clarify the diagnostic values of hematoxylin and eosin (HE), D2-40, CD31, CD34, and HHV-8 immunohistochemical (IHC) staining in gastrointestinal Kaposi's sarcoma (GI-KS) in relation to endoscopic tumor staging. METHODS: Biopsy samples (n = 133) from 41 human immunodeficiency virus-infected patients were reviewed. GI-KS was defined as histologically negative for other GI diseases and as a positive clinical response to KS therapy. The receiver operating characteristic area under the curve (ROC-AUC) was compared in relation to lesion size, GI location, and macroscopic appearances on endoscopy. RESULTS: GI-KS was confirmed in 84 lesions (81.6%). Other endoscopic findings were polyps (n = 9), inflammation (n = 4), malignant lymphoma (n = 4), and condyloma (n = 2), which mimicked GI-KS on endoscopy. ROC-AUC of HE, D2-40, blood vessel markers, and HHV-8 showed results of 0.83, 0.89, 0.80, and 0.82, respectively. For IHC staining, the ROC-AUC of D2-40 was significantly higher (P < 0.05) than that of HE staining only. In the analysis of endoscopic appearance, the ROC-AUC of HE and IHC showed a tendency toward an increase in tumor staging (e.g. , small to large, patches, and polypoid to SMT appearance). D2-40 was significantly (P < 0.05) advantageous in the upper GI tract and for polypoid appearance compared with HE staining. CONCLUSION: The diagnostic value of endothelial markers and HHV-8 staining was found to be high, and its accuracy tended to increase with endoscopic tumor staging. D2-40 will be useful for complementing HE staining in the diagnosis of GI-KS, especially in the upper GI tract and for polypoid appearance.

Endocytoscopic visualization of squamous cell islands within Barrett's epithelium

AIM: To study the endocytoscopic visualization of squamous cell islands within Barrett's epithelium. METHODS: Endocytoscopy (ECS) has been studied in the surveillance of Barrett's esophagus, with controversial results. In initial studies, however, a soft catheter type endocytoscope was used, while only methylene blue dye was used for the staining of Barrett's mucosa. Integrated type endocytoscopes (GIF-Q260 EC, Olympus Corp, Tokyo, Japan) have been recently developed, with the incorporation of a high-power magnifying endocytoscope into a standard endoscope together with narrow-band imaging (NBI). Moreover, double staining with a mixture of 0.05% crystal violet and 0.1% of methylene blue (CM) during ECS enables higher quality images comparable to conventional hematoxylin eosin histopathological images.RESULTS: In vivo endocytoscopic visualization of papillary squamous cell islands within glandular Barrett's epithelium in a patient with long-segment Barrett's esophagus is reported. Conventional white light endoscopy showed typical long-segment Barrett's esophagus, with small squamous cell islands within normal Barrett's mucosa, which were better visualized by NBI endoscopy. ECS after double CM staining showed regular Barrett's esophagus, while higher magnification (×480) revealed the orifices of glandular structures better. Furthermore, typical squamous cell papillary protrusion, classified as endocytoscopic atypia classification (ECA) 2 according to ECA, was identified within regular glandular Barrett's mucosa. Histological examination of biopsies taken from the same area showed squamous epithelium within glandular Barrett's mucosa, corresponding well to endocytoscopic findings. CONCLUSION: To our knowledge, this is the first report of in vivo visualization of esophageal papillary squamous cell islands surrounded by glandular Barrett's epithelium.

The study of selective primary culture and determination of a breast cancer cell line in vitro

Objective The successful establishment of a tumor cell bank is based on the premise that the target cells can be cultured by a legitimate approach.In this experiment,we used primary culture to select and detect breast cancer cells in vitro,which can provide experimental ideas and methods for the establishment of a living tumor tissue cell bank.Methods Fifty-two specimens were collected over a two-year period from people with breast cancer who needed surgical treatment in our hospital.Cells were isolated and used to establish successful cell culture.Cell activity and cell purity were measured before liquid nitrogen cryopreservation.Results(1)At the initial culture stage,cells grew with adherence.Cell multiplication could be seen after the cell medium was exchanged three times.Cell viability was above 86%,while the viability of the target cells was above 75%,as detected by hematoxylin and eosin(HE)staining.(2)The number of breast cancer cells decreased,while the number of fibroblasts increased after five rounds of passage.(3)The success rate was 73.08%,which did not include polluted cells and those that were not successfully cryopreserved.Conclusion(1)breast cancer cells could be selected from primary culture in vitro through an appropriate method.(2)Exchange of the cell medium and further cell passage improved cell multiplication.(3)The experimental results could be monitored using trypan blue and HE staining.(4)The success of breast cancer cell culture in vitro could be used as a reference for other cell culture,so as to establish a tumor tissue cell bank.

Protective effect of liraglutide on the myocardium of type 2 diabetic rats by inhibiting polyadenosine diphosphate-ribose polymerase-1

BACKGROUND In recent years,studies have found that the occurrence and development of diabetic cardiomyopathy(DCM)is closely related to an increase in polyadenosine diphosphate-ribose polymerase-1(PARP-1)activity.PARP-1 activation could be involved in the pathophysiological process of DCM by promoting oxidative stress,the inflammatory response,apoptosis and myocardial fibrosis.AIM To investigate the mechanism of liraglutide in improving myocardial injury in type 2 diabetic rats,further clarified the protective effect of liraglutide on the heart,and provided a new option for the treatment of DCM.METHODS Forty healthy male SD rats aged 6 wk were randomly divided into two groups,a normal control group(n=10)and a model group(n=30),which were fed an ordinary diet and a high-sugar and high-fat diet,respectively.After successful modeling,the rats in the model group were fed a high-glucose and high-fat diet for 4 wk and randomly divided into a model group and an intervention group(further divided into a high-dose group and a low-dose group).The rats were fed a high-glucose and high-fat diet for 8 wk and then started drug intervention.Blood samples were collected from the abdominal aorta to detect fasting blood glucose and lipid profiles.Intact heart tissue was dissected,and its weight was used to calculate the heart weight index.Haematoxylin and eosin staining was used to observe the pathological changes in the myocardium and the expression of PARP-1 in the heart by immunohistochemistry.RESULTS The body weight and heart weight index of rats in the model group were significantly increased compared with those in the normal control group,and those in the intervention group were decreased compared with those in the model group,with a more obvious decrease observed in the high-dose group(P<0.05).In the model group,myocardial fibers were disordered,and inflammatory cells and interstitial fibrosis were observed.The cardiomyopathy of rats in the intervention group was improved to different degrees,the myocardial fibers were arranged neatly,and the myocardial cells were clearly striated;the improvement was more obvious in the high-dose group.Compared with the normal control group,the expression of PARP-1 in myocardial tissue of the model group was increased,and the difference was statistically significant(P<0.05).After liraglutide intervention,compared with the model group,the expression of PARP-1 in myocardial tissue was decreased,and the reduction was more obvious in the high-dose group(P<0.05)but still higher than that in the normal control group.CONCLUSION Liraglutide may improve myocardial injury in type 2 diabetic rats by inhibiting the expression of myocardial PARP-1 in a dose-dependent manner.

Role of Immunostaining in Detecting Extra-Pattern and Subtle Lymphomatous Infiltration in Bone Marrow Biopsies of NHL Patients

Introduction: Immunohistochemistry (IHC) enables the examination of a greater number of trephine biopsy levels and is helpful in determining additional scattered malignant cells. The aim of this study is to detect extra-pattern and subtle lymphomatous infiltration in bone marrow biopsies using CD20 and CD3 immunostaining. Patients and Methods: This study was conducted on 100 newly diagnosed Non Hodgkin Lymphoma (NHL) patients. Their bone marrow trephine biopsies were assessed on routine histology [Hematoxylin and Eosin (H & E)], and were further subjected to IHC using CD20 and CD3. Results: Pattern of involvement by H & E was highlighted by IHC. It showed additional interstitial pattern in 9 cases, parasinusoidal streaks in one case and highlighted a patchy pattern in another case with interstitial involvement on H & E. IHC also detected subtle infiltrations on additional 5.5% cases compared with histology alone. It helped in differentiating reactive (12 cases) and malignant lymphoid infiltration (33 cases). Conclusion: CD20 and CD3 immunostaining performed routinely on bone marrow trephine biopsies has the ability to reveal extra-pattern of infiltration and improve detection of subtle lymphoid involvement. A combined procedure identifying several distinctive features, in particular histotopography and IHC, provides a promising way of discriminating reactive from neoplastic lymphoid infiltrates in bone marrow trephine biopsies.

改良HE染色技术在病理诊断中的应用效果

目的:探讨改良苏木精-伊红(HE)染色技术在病理诊断中的应用效果。方法:选取2021年1月至2022年8月该院182例采用常规HE染色技术的病理组织石蜡切片标本为对照组,另选取同期200例采用改良HE染色技术的病理组织石蜡切片标本为研究组。比较两组染色标准率、标本合格率和不同组织染色有效率。结果:研究组染色标准率、标本合格率均高于对照组,差异有统计学意义(P<0.05);研究组骨组织、脂肪组织、子宫内膜组织、胃镜活检、肝胆组织的染色有效率均高于对照组,差异有统计学意义(P<0.05)。结论:改良HE染色技术应用于病理组织石蜡切片标本诊断可提高染色标准率、标本合格率和不同组织染色有效率,其效果优于常规HE染色技术诊断。

不同固定方法对豚鼠眼球后极部的固定效果比较

目的:为解决豚鼠眼球组织切片制备过程中所存在的视网膜脱片问题,采用不同固定方法,优化固定效果。方法:2周龄正常豚鼠(75只)随机分为5个大组(A-E组),15个小组,每小组5只。A组(1-3小组)眼球分别置于FAS、Davidson固定液1(D1)和Davidson固定液2(D2)中固定24 h;B组(4-6小组)眼球在固定液中固定1 h后剪切角膜,再固定2 h;C组(7-9小组)眼球在固定液中固定1 h后沿视神经方向将眼球分为左右两半,再固定2 h;D组(10-12小组)眼球在固定液中固定3 h后将眼球分为左右两半;E组(13-15小组)眼球在固定液中固定3 h后剪切角膜。经苏木精-伊红(HE)染色比较各个小组眼球后极部固定效果。结果:形态观察表明1-6小组、11-15小组眼球表面光滑圆润,色泽透明,7-10小组眼球凹陷皱缩变形。HE染色表明大部分组别眼球后极部组织切片卷曲缠绕,视网膜脱离;1、5、6、14、15小组切片结构规整,其中14小组形态最佳,视网膜、脉络膜、巩膜连接紧密,组织结构清晰,细胞排列规整。结论:采用D1固定液固定3 h后剪切角膜的固定效果最为理想,适用于豚鼠眼球后极部相关组织研究。

聚羟基丙烯酸和4%中性缓冲甲醛对苏木精伊红染色及荧光原位杂交检测乳腺癌HER-2基因影响的比较

背景与目的：适当的组织固定、透明脱蜡是苏木精伊红（hematoxylineosin，HE）染色及荧光原位杂交（fluorescence加s／tuhybridization，FISH）检测乳腺癌HER．2基因一个重的步骤。该研究旨在对比环保固定液聚羟基丙烯酸和4％中性缓冲甲醛固定制作的组织切片HE染色优良率，荧光原位杂交检测两者固定制作的乳腺癌组织切片的HER-2基因扩增阳性率，探讨其替代传统固定液4％中性缓冲甲醛的可行性。方法：收集中山市博爱医院2011年3月一2015年1月门诊及住院部送检的乳腺癌69例、乳腺纤维腺瘤41例、子宫平滑肌瘤40例、宫颈组织25例、胎盘组织25例，各取材2块，每例标本的2块组织用抽签法随机分2组，一组采用4％中性缓冲甲醛固定制作HE切片200张，另一组采用聚羟基丙烯酸固定制作HE切片200张。依据切片染色情况判定切片等级，采用SPSS19．0比较两者HE染色优良率；两组乳腺浸润性导管癌组织再各制作切片69张，采用SPSS19．0比较FISH技术检测的HER．2基因扩增率。结果：①聚羟基丙烯酸、4％中性缓冲甲醛固定组组织切片HE染色优质片分别为155张、166张，优良片41张、33张，中等片3张、1张，差片1张、O张，染色优良率分别为98．0％、99．5％，两组间优良率差异无统计学意义（x2=1．33，P=-0．25）。②聚羟基丙烯酸、4％中性缓冲甲醛固定组组织切片FISH阳性率分别为26．09％、23．19％，两组间阳性率差异无统计学意义C=0．50，P〉0．05）。结论：聚羟基丙烯酸固定制作的组织切片HE染色效果及荧光原位杂交检测乳腺癌HER．2基因扩增效果跟传统固定液4％中性缓冲甲醛相比差异无统计学意义，符合环保求，有推广使用的可能。

环保透明脱蜡液Van-Clear与传统试剂在组织处理后HE染色及FISH法检测乳腺癌HER2基因中的比较

目的比较环保透明脱蜡液Van-Clear和二甲苯透明脱蜡制作的组织切片苏木精-伊红(HE)染色优良率,对比荧光原位杂交(FISH)法检测2种透明脱蜡液制作的乳腺癌组织切片的人类表皮生长因子受体2(HER2)基因扩增阳性率,探讨Van-Clear替代二甲苯的可行性。方法收集中山市博爱医院2013年2月~2015年12月门诊及住院部送检的乳腺浸润性导管癌标本89例、乳腺增生标本125例、子宫平滑肌瘤标本193例、子宫平滑肌肉瘤标本12例、肺癌穿刺标本16例、绒毛标本139例,同一病变部位切取2个样本,用抽签法随机分为2组,命名为A、B组。A组采用二甲苯透明脱蜡制作切片574张;B采用Van-Clear透明脱蜡制作切片574张。依据切片染色情况判定切片等级,采用SPSS20.0软件比较A、B组切片HE染色优良率;A、B组中乳腺浸润性导管癌标本再各制作切片89张,采用SPSS 20.0软件比较FISH法检测的HER2基因扩增的差异。结果 (1)生物显微镜下,A、B两组切片HE染色结果细胞轮廓清晰、透明度好,细胞核与细胞质蓝红相映、色彩鲜艳,核质对比明显,核膜及核染色质颗粒清晰可见、分辨良好,分裂象染色体呈现黑蓝色,各种成分层次分明。(2)A、B两组切片HE染色优良片分别为570张、572张,中等、差片4张、2张。染色优良率分别为99.30%、99.65%,两组间染色优良率差异无统计学意义(χ2=0.50,P>0.05)。(3)荧光显微镜下,A、B两组切片乳腺浸润性导管癌HER2双探针FISH检测结果组织轮廓和背景均清晰,探针定位准确,可见耀眼的红/绿荧光信号。(4)A、B两组切片检测乳腺浸润性导管癌HER2基因,FISH阳性率分别为24.72%、26.97%,两组间阳性率差异无统计学意义(χ2=0.50,P>0.05),且FISH结果符合率为97.75%。结论环保透明脱蜡液Van-Clear有替代传统试剂二甲苯应用于组织切片HE染色及FISH法检测乳腺癌HER2基因的潜在可能。

病原菌血流感染对阿尔茨海默病小鼠记忆力和海马锥体细胞的影响

目的 探讨病原菌血流感染对阿尔茨海默病(AD)小鼠记忆力和海马锥体细胞的影响。方法 50只C57BL/6J雄性小鼠腹壁皮下注射D-半乳糖液[200 mg/(kg·d)]连续8周构建AD模型并于造模期间每周观测小鼠的一般状态及体质量;造模结束后均分为生理盐水(对照)组、大肠埃希菌组、铜绿假单胞菌组、肺炎克雷伯杆菌组、金黄色葡萄球菌组及白念珠菌组,对照组小鼠尾静脉注射生理盐水,其余病原菌感染组小鼠尾静脉注射低(1.5×107CFU/L)、中(1.5×108CFU/L)及高(1.5×109CFU/L)浓度的对应病原菌菌液,观察14 d,分别于注射后第1、2、3天及第1、2周测定各组小鼠的体质量及体温;观察结束后,于第15天采用Morris水迷宫实验检测各组小鼠逃逸潜伏期、游泳速度及穿越平台次数,水迷宫实验共进行6 d;实验结束后处死各组小鼠,取海马组织,采用苏木精-伊红(HE)染色观察海马组织学的改变。结果 小鼠于第8周体质量开始下降,且精神状况较造模初期明显下降;部分病原菌感染组小鼠逃避潜伏期较对照组延长(P<0.05),第5天肺炎克雷伯菌高浓度组小鼠水迷宫实验游泳速度较对照组下降(P<0.05);对照组小鼠海马CA1和CA3区神经细胞及皮层未见明显异常,白念珠菌组小鼠海马组织中发现少量凋亡细胞,肺炎克雷伯菌组、金黄色葡萄球菌组小鼠海马神经细胞轻度排列紊乱。结论 AD患者来源的病原菌经尾静脉血流感染可导致AD小鼠记忆力下降及海马锥体细胞的形态改变。

HE切片褪色后行EBER原位杂交检测方法的探讨

目的探讨将HE切片褪色后行EB病毒编码的小RNA(EBER)原位杂交检测的方法。方法收集40例EBER原位杂交检测阳性病例的HE切片作为实验组,相同蜡块的连续切片作为对照组,分别进行EBER原位杂交检测。结果实验组中,38张切片的组织结构全部完整,2张骨髓穿刺切片中的骨小梁脱片,但剩余组织组织结构保持完整。所有40张切片均杂交阳性,阳性信号位于细胞核,呈棕黄色,定位准确、清晰可读,杂交背景干净。阳性细胞数目及强度与对照组无差别。结论将HE切片褪色后行EBER原位杂交检测,方法简单、可靠,在病理诊断中有一定的应用价值。

HE切片中苏木素沉渣去除方法探讨

目的:探讨去除HE染色切片中苏木素沉渣的方法。方法:采用下列三种方法去除HE染色切片中的苏木素沉渣并比较其效果:(1)陈旧苏木素染色液加热至40℃;(2)滤纸过滤陈旧苏木素染色液;(3)石蜡切片经1.0%NaOH浸泡8min、1.0%NaHCO3浸泡2min、200U/ml糜蛋白酶浸泡8min后在陈旧苏木素染色液中染色。此外,在AB/PAS染色前和染色后用1.0%NaOH、1.0%NaHCO3、200U/ml糜蛋白酶浸泡切片,观察该化学处理对粘液染色效果的影响。结果:经过1.0%NaOH、1.0%NaHCO3或200U/ml糜蛋白酶浸泡的切片,HE染色后无苏木素沉渣附着;陈旧苏木素染色液加热后染色,切片中仍有大量苏木素沉渣附着,过滤后染色则切片中苏木素沉渣消失。在AB/PAS染色后、苏木素衬染前1.0%NaOH、1.0%NaHCO3或200U/ml糜蛋白酶处理切片,粘液区呈强阳性染色;AB/PAS染色前用上述方法处理切片,粘液染色强度略有减弱。结论:陈旧苏木素染色液过滤经1.0%NaOH、1.0%NaHCO3、200U/ml糜蛋白酶浸泡均可有效去除HE染色切片中的苏木素沉渣。

不同脱钙液对牙齿牙周联合石蜡切片HE染色的影响

目的探讨不同脱钙液对牙齿牙周联合切片HE染色质量的影响。方法选用10%乙二胺四乙酸钠(EDTA)、复合酸(Plank-Rychlo脱钙液)和5%硝酸3种不同的脱钙液,分成A、B、C3组用于犬和大鼠牙齿牙周组织联合石蜡切片脱钙,通过记录脱钙时间,观察HE染色效果,评价室温下(20℃～28℃)不同脱钙液对切片制作的影响。结果A组脱钙液所制切片染色效果良好,但脱钙速度缓慢,所需时间较长;B组脱钙液脱钙速度快,所制切片染色好;C组脱钙液所制切片的染色强度和对比度均不如前两种脱钙液。结论在制作牙齿牙周联合切片时,EDTA脱钙液和Plank-Rychlo脱钙液所制切片均能取得良好的染色效果,但EDTA脱钙液脱钙周期过长;Plank-Rychlo脱钙液脱钙均匀,速度快,更适合于临床速检工作;硝酸脱钙液组染色效果较差。

组织染色中HE染色法的改良

目的:比较改良苏木素伊红(HE)染色法与传统染色法的染色效果。方法:分别用传统染色法和改良染色法对组织切片和细胞涂片进行染色,进行比较。结果:改良染色法较传统染色法,染色液中沉淀更少,无氧化膜,染色时间短,不需过滤,使用期限长,染色效果好。结论:改良染色法克服了传统染色法的缺点,有利于切片质量的提高。

环保试剂聚羟基丙烯酸和Van-clear用于HE染色效果分析

目的探讨聚羟基丙烯酸和Van-clear在苏木精-伊红染色法(HE染色)中的应用效果。方法收集河北省人民医院2018年1月至2019年1月期间送检的脑组织(12例)、脾组织(18例)、骨髓组织(22例)、淋巴结组织(50例)、淋巴瘤组织(10例)、乳腺纤维腺瘤组织(62例)、宫颈腺癌组织(26例)、子宫平滑肌瘤组织(74例)、慢性宫颈炎组织(102例)、胎盘组织(35例)、脂肪组织(57例),同一部位各取材两块,采用抽签法随机分为A组和B组。A组采用传统甲醛和二甲苯处理HE切片468张,B组采用环保试剂聚羟基丙烯酸和Van-clear处理HE切片468张,观察对比两组切片HE染色情况。结果两组切片均取材完整、厚度适宜、未见褶皱或刀痕、HE染色核浆清晰、红蓝适度、透明洁净、封裱美观;A组优良率与B组优良率相比差异无统计学意义(98.08%比98.71%)(P>0.05);将A组HE染色结果作为参照,B组切片的优、良、中、差染色结果与A组染色结果的符合率分别为97.4%(456/468)、98.2%(459/468)、99.4%(465/468)、100%(468/468),本研究符合率最小为97.4%,将A组HE染色结果作为参照,B组HE染色结果预期符合率较高。结论聚羟基丙烯酸和Van-clear处理切片相较于甲醛和二甲苯更加环保、对病理工作人员的刺激性小、安全性高,在不改变任何处理程序的情况下效果相当,对于病理实验室而言具有运用推广意义。