Exploration of Methods for Preparation of Cattle Eyeball Paraffin Sections

[Objective] This study aimed to establish a new method for preparing paraffin sections of cattle eyebal s. [Method] The conventional method was used to prepare paraffin sections for cattle eyebal s in the control and a new method termed＆quot;opening a window on cornea and refixation＆quot; was used to prepare paraffin sections for cattle eyebal s in the treatment group. [Result] After the prepared specimens in the treatment group were fixed, it could be macroscopical y observed that retina and choroid were closely connected, with detachment occurring at a smal portion be-tween the two. According to the paraffin sections, it was microscopical y observed that the continuity of trabecular meshwork was intact, as wel as the continuity be-tween different layers of eyebal wal , without detachment between them, no retinal detachment, no shrinkage of each layer of tissue cells. [Conclusion] This study pro-vides a foundation for the basic research and pathological study of eyebal s.

Paraffin section observation of flower bud differentiation of Chimonanthus praecox in Kunming and comparison of the differentiation processes in different regions,China

Chimonanthus praecox is an important ornamental plant and cut flower material in China.It blooms in the freezing winter and its flower emits charming fragrance.However,in different region the flowering time is variable.In order to understand the flowering mechanism of Ch.praecox in the winter,we studied the flower bud differentiation in Spring City-Kunming using paraffin sectioning method in the present study.Meanwhile we compared the differentiation process difference from different regions.It was found that the temperature is the key factor for its flower bud differentiation and blossom of Ch.praecox.In the process of bud differentiation,the temperature 20℃was the optimum for inducing changes from vegetative axillary buds to reproductive buds and subsequent morphological differentiation in Ch.praecox.Furthermore in the first three differentiation periods—tepal primordial stage,staminal primordial stage and pistil primordial stage,Kunming took the shortest time to finish the process due to very rapid temperature rise to 20℃,whereas,in Zhengzhou the time for these differentiations was the longest,which may be caused by the slow temperature rise.After May,the high temperature stress forced the flower buds into the first long dormant period in all regions except Kunming.In Kunming,the average temperature was only 20–25℃,so the flower bud continued to differentiate.In all regions,Kunming is the first to complete whole flower bud differentiation even on the early August,and started the second dormancy very early but very long.In the other regions,the plants went through a shorter dormancy and the low temperature broke the dormancy rapidly.Contrarily the plants of Kunming spent a longer period for the low temperature.Thus,the low temperature less than 10℃is a key factor to breaking the second dormancy.Surely the regular effects of temperature on flower bud differentiation and blossom is very helpful for florescence regulation of Ch.praecox.

Preparation of Paraffin Section of Pinus sylvestris Needles

[Objective] Taking Pinus sylvestris needles as materials, a set of test con-ditions suitable for the preparation of paraffin sections of Pinus sylvestris needles was determined. [Method] Based on the traditional method for paraffin sections preparation, steps including fixation, dehydration, adhering to slides and staining were investigated taking the structural characteristics of pine needles into considera-tion. [Result] 70% ethanol was used in the FAA fixative; before affixed on slides, the cut sections were first expanded in a 40 ℃ water bath and taken out by using Su-perfrost Plus slides, fol owed by drying in a 35 ℃ oven for 24 h; pine needles were cut into 8 μm thick sections; staining was achieved by immersion in eosin for 5 min and in hematoxylin for 1 min. [Conclusion] The conditions obtained above can en-hance the effect of fixation and dehydration; adhering to slides is easy to operate and the sections are not easy to drop; the dyeing effect is relatively preferable.

Production Method for Paraffin Section of Invasive Species of Bemisia tabaci

[Objective] The purpose was to introduce a preparation method for paraffin section of Bemisia tabaci, so as to lay the foundation for the studies on changes of organizational structure of B. tabaci. [ Method] The technique of paraffin section and the method of H-E staining were adopted to study the organizational structure of B. tabaci. The slices were examined and photographed under fluorescence microscope. [ Result] The coloring of H-E staining cells was good. Under 400 x conditions, the fat body, compound eyes, nucleus and muscle of B. tabaci were clear. [ Conclusion] The production method for paraffin section of B. tabaci was reliable, and the quality of slices was high.

RAPID DETECTION OF ONCOGENE AMPLIFICATION IN PARAFFIN SECTIONS OF BREAST CARCINOMAS USING QUANTITATIVE DIFFERENTIAL PCR AND FLUORESCENT DNA TECHNOLOGY

Gene amplification is a common mechanism of oncogene activation and contributes to tumor progression. Analysis of such genetic alterations are relevant to the understanding of tumor genetics and could provide prognostic information for the individual patient. Standard analytical approaches using Southern blot and slot blot require a large amount of good

一种优化的桑树花芽石蜡切片制作方法

针对常规石蜡切片方法无法获得完整桑树花芽切片的问题,以“无籽大十”品种桑树花芽为供试材料,在常规石蜡切片技术基础上,对石蜡切片样品制作过程中固定、软化、脱水、透明及染色等环节进行了优化。结果表明,使用15%氢氟酸软化后,采用优化后的系列脱水、透明等步骤进行制片,可以使切片连续性、完整性得到明显提高。通过切片显微观察,可见细胞轮廓明显、裂痕较少、结构较完整,染色较清晰,并且对发育后期的花芽效果更为明显。优化后的制样流程解决了常规制片出现浸蜡不彻底、组织变脆、蜡带不连续、内部结构不完整等一系列问题。该研究可以从解剖学上为桑树花芽发育形态和花芽分化机制研究提供技术参考。

二甲苯、正庚烷和正丁醇对脂肪组织透明效果的比较研究

为了找到安全、高效、可替代二甲苯的脂肪组织石蜡切片透明剂,试验采集2月龄健康民猪背部脂肪组织,经固定、脱水后,分别用二甲苯、正庚烷、正丁醇处理不同时间(二甲苯处理1 h,正庚烷处理1 h、2 h、3 h、4 h,正丁醇处理1 h、2 h、3 h、4 h、10 h)进行透明,观察透明效果,然后进行石蜡包埋和切片;选取3种透明剂将脂肪组织完全透明用时最短的石蜡切片,经脱蜡和水化处理后用苏木精染液和伊红染液进行H.E.染色,探究3种透明剂对脂肪组织形态结构的影响;并采用免疫组织化学技术检测过氧化物酶体增殖物激活受体γ(peroxisome proliferators-activated receptor γ,PPARγ)的表达情况,探究3种透明剂是否会影响蛋白质的表达和定位。结果表明:二甲苯处理1 h、正庚烷处理2 h后脂肪组织可完全透明,而正丁醇处理10 h后脂肪组织才完全透明;正庚烷、正丁醇作为透明剂制备的脂肪组织石蜡切片细胞形态清晰、结构完整、着色好,与二甲苯作为透明剂的效果相似;在3种透明剂制备的脂肪组织石蜡切片中,PPARγ均清晰且准确地表达在脂肪细胞的细胞核中。说明正庚烷和正丁醇对脂肪组织的透明效果与二甲苯相似,但正庚烷透明速度较快,正丁醇透明速度很慢,因此实际应用中推荐使用正庚烷作为二甲苯的替代物。

制样方法对菊花花蕾激光显微切割样品RNA提取质量的影响

[目的]研究不同固定剂及切片制样方式对菊花花蕾激光显微切割样品RNA提取质量的影响。[方法]以丙酮和乙醇冰醋酸2种试剂固定菊花花蕾,采用石蜡和冰冻切片2种方式制作样本,显微切割并收集最外两轮小花原基,提取RNA后比较提取质量。[结果]石蜡和经梯度蔗糖保护的冰冻切片均能很好地保存花蕾组织形态,就石蜡切片而言,乙醇冰醋酸固定组织切割样品的RNA降解明显低于丙酮固定样品;而2种固定剂的冰冻切片切割样品RNA提取质量无明显差异。与石蜡切片相比,冰冻切片显微切割样品RNA质量更高。切割时收集管盖加入RNA提取缓冲液可明显缓解RNA降解。显微切割的不同品种菊花花原基与花发育有关的基因表达水平存在差异,表明切割获得的材料可用于下一步研究。[结论]植物组织经乙醇冰醋酸固定、梯度蔗糖保护后冰冻切片进行显微切割效果较佳,可富集特异组织或细胞并从中获得较高质量的RNA。

术中冰冻病理诊断对眼睑肿物切除的价值探讨及病理类型分析

目的 探讨送检术中冰冻病理诊断(frozen sections, FS)的眼睑肿物病理类型及其对眼睑肿物和切缘的诊断准确率,分析FS与石蜡病理诊断不一致的常见原因。方法 选取2016年12月至2021年12月首都医科大学附属北京同仁医院送检FS眼睑肿物标本561份,分析眼睑肿物的病理类型,并以石蜡病理诊断为金标准,分析FS对眼睑肿物和切缘的诊断准确率。结果 561份标本中,左眼280份,右眼281份;上睑225份,下睑311份,内眦18份,外眦7份;肿物最大直径1~35 mm,平均(15.1±9.4)mm。其中良性病变165份(29.4%),位居前三位的分别为炎性病变、色素痣和脂溢性角化病;恶性病变396份(70.6%),位居前三位的分别为基底细胞癌、皮脂腺癌和鳞状细胞癌。FS对肿瘤和切缘的诊断准确率分别为97.1%、99.3%,肿瘤延迟诊断率为6.2%。肿瘤诊断不一致类型为误分类(60.0%,9/15)、假阴性(26.7%,4/15)及假阳性(13.3%,2/15)。诊断不一致的原因为解读失误(73.3%,11/15)、制片失误(26.7%,4/15)。结论 送检FS的眼睑肿物以恶性病变为主,并以基底细胞癌和皮脂腺癌多见;FS对眼睑肿物及切缘均具有较高的诊断准确率,能很好地指导手术开展,皮脂腺癌和恶性肿瘤的特殊亚型是与石蜡病理诊断不一致的主要因素,切片深度不够是导致漏诊和诊断不充分的主要原因,日常工作中对于可疑复发及较小的肿瘤应给予充分评估。

小鼠整胚石蜡切片制备方法的改进

Objective:The morphology analysis of whole-mount mouse embryos was observed using an improved paraffin section technique.Methods:Mouse embryos of varying embryonic ages were collected and whole-mount embryo paraffin sections were prepared using PFA-intravenously injected fixation,prolonged dehydration,and paraffin embedding.Hematoxylin and eosin(H&E)staining and immunohistochemical staining were employed to evaluate the quality of sections,with different tissues being observed labeled by CD34.Results:Following a series of tissue processing and staining procedures,the structure of the whole-mount mouse embryo was well-preserved,and the staining was clear and easily distinguishable.Embryos of different embryonic ages were treated differently,yet the quality of tissue processing remained highly consistent.Conclusion:Tissue processing and staining have been significantly improved,allowing for the easy acquisition of whole-mount mouse embryos of different ages through simplified methods of tissue fixation and dehydration duration.The staining results are clear and stable,providing technical support for the study of mouse embryo development.

锦红冰糖橙大果芽变果实结构解剖、激素变化和转录组分析

【目的】探究锦红冰糖橙大果芽变的形成原因。【方法】以锦红和大果芽变为试材,采用石蜡切片、HPLC-MS/MS和转录组测序技术对其在细胞形态、激素含量和基因表达方面进行比较分析。【结果】与锦红相比,大果芽变果肉内汁胞数量增多,且汁胞变大。石蜡切片结果表明,大果芽变汁胞内细胞数量多于锦红。大果芽变内生长素和赤霉素含量在花后70和120 d均高于锦红,玉米素含量在花后70、120和170 d均低于锦红。对盛花后70、120和170 d的汁胞进行转录组分析得到4597个差异基因,其中24个激素信号相关基因在两个及以上时期差异表达。在大果芽变中有4个生长素相关基因在花后70 d上调表达。细胞分裂素相关基因ARR2在大果芽变花后70和120 d均下调表达。细胞周期蛋白CycD3在大果芽变花后170 d上调表达。【结论】锦红冰糖橙大果芽变的变异可能与汁胞发育有关,并且受激素水平影响。转录组分析为解析锦红冰糖橙大果芽变的分子机制奠定了基础。

光周期处理对刺槐无性系组培苗生根的影响

【目的】研究不同光周期对刺槐无性系组培苗不定根发生的影响,分析不同光周期下根原基突出时间及不同阶段根系的生长状态和生理指标,探讨其生根能力,筛选出促进刺槐生根的最适光周期。【方法】以4个二倍体刺槐无性系组培苗为材料,设置5种光周期处理,统计分析不同光周期下刺槐生根率及生理指标变化,并观察根外部形态变化,同时每天取根基部制作石蜡切片,观察根原基发生时间。【结果】1)解剖学观察显示,刺槐无性系组培苗生根过程分为3个时期:愈伤组织形成期(0~7 d)、不定根形成期(根原基诱导期)(7~10 d)、不定根伸长期(10 d后)。2)生根情况及解剖学观察结果显示,4个刺槐无性系中,D26-5①刺槐无性系根原基较早出现,同时生根能力最强,生根率为92%,且24 h/0 h光周期处理下根原基突出时间最早,生根率提高至97.8%。3)生根过程中,可溶性糖和叶绿素含量随着光周期增加而上升,24 h/0 h光周期下含量最高。丙二醛含量呈现出下降—上升—下降的趋势,脯氨酸含量呈先上升再下降的趋势,两者在24 h/0 h光周期处理下的含量都要小于16 h/8 h光周期,说明刺槐应对连续光照时可通过调节渗透物质等方式适应环境的变化。【结论】刺槐无性系组培苗不定根发生能力受光周期影响,在24 h/0 h光周期下根原基突出时间最早,生根能力最强。

沙冬青适应干旱环境的解剖结构变化

以沙冬青盆栽苗为试材,采用石蜡切片法,研究了不同土壤含水量对其根、茎、叶解剖结构的影响,以期为沙冬青的开发利用提供参考依据。结果表明:沙冬青在土壤含水量为2%~4%时仍能正常生长,但不同含水量下沙冬青根、茎、叶结构存在差异。随土壤水分的降低,沙冬青根系生长越细长,木质化程度越深,周皮相对厚度增大,通过发育分化根部结构来提高植株抗旱性。沙冬青茎和叶表皮外附角质层,茎皮层内有皮层维管束提高输导能力,同时在土壤水分低时髓依旧较发达,表现出较强的贮水能力。沙冬青叶为等面叶,随土壤水分的降低叶片厚度、角质层厚度、孔下室深度、栅栏组织/海绵组织的比值、主脉维管组织、木质化程度均呈不同程度增加,通过降低水分散失、提高气体交换、提高叶面积利用率和光合作用来适应干旱环境。

免疫荧光检测石蜡切片IgA蛋白在诊断疱疹样皮炎和线状IgA大疱性皮病的应用评价

目的:评价直接免疫荧光染色检测石蜡切片IgA抗体对于诊断疱疹样皮炎(DH)和线状IgA大疱性皮病(LABD)的应用价值。方法:选取2019-2024年我院病理诊断的DH和LABD病例,入选实验组病例DH 46例,LABD 32例。采用FITC荧光标记兔抗人IgA抗体进行直接免疫荧光染色检测。结果:46例DH石蜡切片中,40例IgA在真皮乳头为阳性,阳性率86.96%。32例LABD石蜡切片中,20例IgA在表皮基底膜线状沉积为阳性,阳性率62.50%。结论:直接免疫荧光染色检测石蜡切片IgA可以作为辅助诊断DH和LABD的有效方法。

槲树不同发育时期胚珠石蜡切片制备体系的优化

【目的】建立并优化适宜槲树子房或胚珠石蜡切片的制作体系,为后续槲树胚胎发育学研究提供技术支撑。【方法】以不同发育时期的槲树雌花为试验材料,将材料以60 d为界分为60 d前和60 d后两部分,对材料处理方式(授粉60 d前后的材料分别进行仅保留子房与胚珠处理)、浸蜡时间(18,24,36 h)与浸蜡条件(60℃烘箱内抽真空处理0,6,8,12 h)、切片厚度(6,8,10μm)进行优化;在此基础上,以授粉70 d样品切片为材料,对染色方法(染色液分别为5 g/L苏木精、1 g/L甲苯胺蓝和10 g/L番红-固绿)及染色时间(1,2,5,10,15 min)进行优化,建立适合槲树胚珠的石蜡切片制备体系。【结果】授粉60 d前的样品仅保留子房部分,授粉60 d后的样品仅保留胚珠,两者在50℃烘箱经甘油乙醇浸泡48 h的软化处理后,所得切片效果最佳,可避免蜡带破裂等不良现象出现。将浸蜡时间延长至36 h,并于第1次浸纯蜡过程中在60℃烘箱内先进行12 h抽真空处理,可保证发育后期体积较大材料的充分浸蜡;脱蜡前将玻片进行3 h抽真空处理,可明显降低脱片率;授粉60 d前样品的切片厚度以8μm为宜,授粉60 d及以后的样品以10μm最佳。槲树胚株石蜡切片制备时,采用5 g/L苏木精染色5 min时的制片效果最好。【结论】建立并优化了槲树不同发育时期子房或胚珠石蜡切片的制备体系,采用该优化体系可获得蜡带连续、结构完整、染色均匀、背景清晰的槲树子房或胚珠切片。

外源2,4-表油菜素内酯及褪黑素对樱桃番茄果实品质和果皮形态结构的影响

【目的】研究不同浓度的2,4-表油菜素内酯(EBR)和褪黑素(MT)处理对樱桃番茄果实生长发育过程中的果实品质和果皮形态结构的影响,为提高樱桃番茄果实品质提供一定理论或技术支撑。【方法】以樱桃番茄果实为材料,在樱桃番茄膨大期、绿熟期、转色期、红熟期喷施3种浓度的EBR和MT,分析果实的品质和细胞结构。【结果】累计喷施植物生长调节剂,在果实进入红熟期均产生不同程度的影响,单果质量在100μmol/L的MT处理下显著高出对照组29.96%;果实硬度在0.1 mg/L的EBR处理下与对照组相比提升了17.21%;0.1 mg/L的EBR和100μmol/L的MT分别是2种植物生长调节剂中糖酸比、可溶性蛋白、VC相比于对照(CK)提升最高的,分别提升了51.14%和58.81%、37.96%和39.40%、49.05%和27%;0.05 mg/L的EBR和100μmol/L的MT分别是2种植物生长调节剂中可溶性固形物提升最高的,分别提升了24.01%和44.10%;3种EBR处理下外表皮与CK相比均有所提高;厚角组织在0.1 mg/L的EBR处理下与对照组相比增加了10.06%。【结论】0.1 mg/L的EBR处理下的樱桃番茄的耐贮藏性和运输性较佳,100μmol/L的MT处理下的樱桃番茄内在品质更佳。

电磁引导经会阴前列腺穿刺联合小标本快脱病理学检查在前列腺癌诊断中的应用价值

目的探讨电磁引导局麻经会阴前列腺mpMRI-TRUS多模态影像融合穿刺活检联合小标本快脱病理学检查在前列腺癌诊断中的应用价值。方法回顾性分析138例PI-RADS评分≥3分并行电磁引导局麻经会阴前列腺多模态影像融合穿刺活检患者的临床病历资料。使用AI技术将mpMRI与TRUS多模态影像进行融合,对可疑病灶靶向穿刺2针,然后行前列腺系统穿刺12针。靶向穿刺标本进行小标本快脱病理学检查,比较靶向穿刺、系统穿刺及联合穿刺csPCa检出率的差异。结果138例患者csPCa检出率为71.01%。靶向穿刺和系统穿刺csPCa检出率分别为62.32%和70.29%,差异无统计学意义(P=0.20)。联合穿刺csPCa检出率高于靶向穿刺和系统穿刺,但差异均无统计学意义(P>0.05)。前列腺PI-RADS评分3分患者靶向穿刺、系统穿刺和联合穿刺的csPCa检出率分别为30.95%、38.10%和40.48%;评分4分患者分别为52.94%、61.76%和61.76%;评分5分患者分别为90.32%、96.77%和96.77%,不同PI-RADS评分患者三种穿刺方法在csPCa检出率方面差异均无统计学意义(P>0.05)。前列腺PI-RADS评分3、4、5分患者,靶向穿刺csPCa漏诊率分别为23.53%、14.29%和6.67%。结论对于PI-RADS v2.1评分≥3分的可疑前列腺癌患者,电磁引导经会阴前列腺穿刺活检联合小标本快脱病理学检查可快速获取病理结果,且有较好的诊断准确性,但联合穿刺仍是目前诊断前列腺癌最合适的方法。

佛罗里达文昌鱼哈氏窝组织在性腺发育不同时期的形态结构变化

哈氏窝是文昌鱼生殖内分泌调控轴重要组成部分,为了进一步揭示文昌哈氏窝形态特征与性腺发育的关系,丰富文昌鱼的生物学基础信息,利用组织学切片及HE染色技术观察了佛罗里达文昌鱼在性腺发育不同时期哈氏窝形态变化。结果表明:佛罗里达文昌鱼在性腺刚出现及早期时,哈氏窝位于文昌鱼哈氏窝位于脊索附近,有一层薄膜相隔,呈近似对称的倒“凹”状;在性腺发育和成熟时期,哈氏窝顶部增厚,胞质清晰,两侧向窝内增大,窝内空隙变小,形状呈现矮宽;在性腺排空及退化时期,哈氏窝两侧呈细长状,窝内空间狭窄。

萱草愈伤组织分化的研究

为了解决萱草愈伤组织仅生长增大并未分化出芽的问题,提高其繁殖系数,采用不同浓度配比的6-BA和IBA为外源激素,对选育的JH2-2萱草继代培养的愈伤组织接种培养,并对不同颜色的愈伤组织制成石蜡切片进行观察。结果表明:在接种培养的第6天开始有小绿芽自愈伤组织上分化而出,第21天愈伤组织的分化率达到最大值,同时部分愈伤组织出现玻璃化,且后续没有分化发芽迹象。外源激素6-BA比IBA对萱草愈伤组织的分化影响更大。适合萱草愈伤组织进行分化培养的培养基比例组合为:MS+蔗糖30 g·L^(-1)+琼脂6 g·L^(-1)+6-BA 1.0 mg·L^(-1)+IBA 0.2 mg·L^(-1),pH值为6.5,分化率达100%。对不同颜色的萱草愈伤组织制成石蜡切片进行观察,结果发现其中一种为胚性愈伤组织,外形呈球形颗粒状,细胞排列整齐、紧密,细胞核染色较深;另一种为非胚性愈伤组织,外形松软粗糙无固定形状,细胞大小不均匀且无固定形状,细胞核不明显。在萱草的组织培养过程中,可以优先考虑接种乳白色和绿色的活性高的愈伤组织,对于提高萱草的分化率和繁殖率具有重要意义。本研究结果可为优化萱草离体快繁技术体系,进一步创造新品种提供一定的技术支撑。

术中冰冻切片对甲状腺乳头状癌的诊断准确性分析

目的探讨术中冰冻切片(FS)诊断甲状腺乳头状癌(PTC)的准确性和价值,并对提高诊断准确性的措施进行思考。方法选择40例疑似PTC患者,均进行术中FS病理诊断、石蜡切片病理诊断、B超引导下的细针吸取细胞学(FNAC)检查。以石蜡切片诊断结果作为金标准,分析术中FS的诊断准确性、敏感性和特异性。结果40例患者经石蜡切片诊断后共有39例确诊为PTC,将其作为金标准;经术中FS诊断后共有38例确诊,其中仅有1例漏诊,FS诊断准确率为97.50%(39/40),敏感性为94.74%(38/39),特异度为100.00%(1/1)。B超报告病灶为3、4、5类甲状腺结节;术中FS对3、4、5类甲状腺结节的诊断准确率分别为100.00%、92.31%、100.00%。结论术中FS在PTC的诊断中可以取得与石蜡切片相对一致的结果,而且术中FS所谓时间短,通常不超过30 min,操作简单,更具实用性。