Air drying as a preparatory factor in cytology: investigation of its influence on dye uptake and dye binding

The effect of air-drying on cytological material is investigated in this article. Smears of rat liver were fixed completely wet and, after air drying, postfixed in ethanol, methanol/formaldehyde/acetic acid (MFA), and formaldehyde. Staining was performed with the thionin-Feulgen procedure, a standard Romanowsky-Giemsa stain with azure B-eosin Y and a Papanicolaou staining variant. The image analysis system IBAS 2000 was applied to evaluate objective criteria of the changes caused by air drying the chromatin texture. Nuclear absorption was measured with a Vickers M 85a Microdensitometer. Air-drying had striking effects on size and shape of cell nuclei (spreading), on the structure of the nuclear chromatin (chromatin condensation), and on the chromaticity coordinates (hue, saturation, and intensity of nuclear staining). The variations of the chromatin texture and dye-substrate affinity are attributed to alterations of the tertiary structure of the nuclear proteins.     

细胞化学染色在急性白血病分型中意义的探讨

目的 采用形态学及细胞化学染色方法对78例急性白血病进行诊断分型。方法 按照FAB标准进行形态学分型，采用Wright-Giemsa染色；细胞化学染色包括过氧化物酶，非特异性酯酶即α－醋酸萘酚酯加氟化钠抑制，糖原即过碘酸雪夫染色。结果 过氧化物酶、糖原反应可区别急性淋巴细胞白血病与急性非淋巴细胞白血病，同时糖原反应对诊断M6具有特异性；非特异性酯酶加氟化钠抑制试验，对区别粒－单白血病具有重要意义。细胞化学染色与形态学分型符合率为89.4%，形态学的确诊率为74.4%，加做细胞化学染色可将诊断率提高到93.5%。结论 形态学是白血病诊断分型的基础，细胞化学染色是形态学的重要补充，其经济、简便和稳定的特点有利于广大基层医院的开展。     

Small megakaryocytes — an identification problem

The number of bone marrow megakaryocytes (MK) was estimated in May Grunwald-Giemsa-stained (MGG) and immunoenzymatically stained smears. The latter was performed after incubation with a monoclonal antibody (ITI-PL1) directed against blood platelets and megakaryocytes (MK). The MK were grouped according to their size (diameter above or below 30 micron, respectively). Altogether 7.9% more MK were recognized in ITI-PL1 stained smears as compared to MGG-stained smears, mainly because comparatively few small MK were recognized in MGG-stained smears.     

海南省16个疟区市县“三热”病人血片抽查复检结果分析

目的了解基层疟原虫镜检人员的镜检技术、血片制作和染色水平,提高镜检质量,为海南省2018年前实现消除疟疾目标提供科学依据。方法 2008～2010年对琼海市、琼中县等16个市县＂三热＂病人血片抽样复检。结果复检血片21 817张,复检符合率为99.3%;其中阴性血片和阳性血片复检符合率分别为99.99%、93.14%;血片制作和染色优中等级血片分别占92.33%和88.89%。结论镜检符合率都达到了较高水平,但漏检、假阳性、虫种错检及染色效果不佳的问题仍然存在。因此,有针对性的加强基层镜检人员的培训,提高其镜检技术、血片制作和染色水平,以及镜检人员的责任心,是目前疟防工作的重要内容之一。     

The effects of fixation,processing and evaluation criteria on immunohistochemical detection of hormone receptors in breast cancer

A task force of the Japanese Breast Cancer Society has proposed a recommendation for adequate evaluation of hormone receptors in routine practice, in order to standardize handling of tissues, staining techniques and scoring systems. As a part of the study, several examinations were conducted to detect the effect of technical problems, including the influence of fixation time and other fixation and processing conditions, on the immunoreactivity for ERalpha. There is little influence of prolonged fixation on the immunoreactivity for ERalpha, except for cases in which particularly over-fixed blocks are used. A delay in the onset of fixation could decrease the immunohistochemical findings of steroid receptors, compared with shorter or longer fixation, and the situation is similar to the fixation of a whole large surgical specimen in formalin in a big bucket. Incomplete fixation might be an important cause of heterogeneiety of immunoreactivity for ERalpha. Manual and automated immunohistochemical (IHC) staining by DAKO (Glostrup, Denmark) and Biogenex (San Ramon, CA) and automated IHC staining by Ventana Medical Systems (Tucson, AZ) each employ different methods. Using a scoring system, in which the proportion of cells stained in each specimen was recorded as 0, less than 1%, 1% or more and less than 10%, and 10% or more, the intermethod variability of those IHC staining methods exhibited substantial multi-rater kappa values concerning the ER and PgR (kappa for ER according to the percentage of positive cells=0.67; PgR=0.72). Concerning intermethod consistency, the scoring system based on the percentage of positive cells was advantageous over other scoring systems, based on the intensity of nuclear staining. Using double staining, patients with ER-positive and HER2-positive tumors can be classified as those with co-expressed tumors and those with differently expressed tumors. As such, the co-expressed tumor might be resistant to antiestrogen therapy in ERalpha-positive and HER2-positive breast cancer and double staining might lead to the development of new therapeutic strategies for hormone and HER2-positive breast cancer.     

斑点免疫金银染色法诊断囊虫病的研究

用斑点免疫金银染色法(Dot-IGSS)和Dot-ELISA检测40份确诊为囊虫病患者的血清。两种方法检测的阳性率均为100％,用Dot-IGSS检测,40份血清的平均滴度为1:27470,显著高于用Dot-ELISA检测的平均滴度1:9069(P<0.01)。检测50份义务献血员血清,假阳性率分别为2％和4％。10份日本血吸虫病人血清分别有1份和2份出现假阳性。与肝吸虫病人血清无交叉反应,与包虫病人血清有明显的交叉反应。     

吞噬细胞吞噬作用实验方法的改进

目的探讨和改进实验方法,以便更清晰、有效地观察吞噬细胞对外来异物的吞噬作用。方法小吞噬试验是用金黄色葡萄球菌感染昆明小鼠、大吞噬试验是将鸡红细胞注入Hartly豚鼠体内,然后取鼠腹腔液涂片、甲醇固定,分别用瑞氏染色法和瑞氏-姬姆萨染色法进行染色,镜下观察吞噬细胞对外来异物的吞噬作用。结果瑞氏-姬姆萨染色法制作的标本比瑞氏染色法制作的标本显微镜下观察吞噬细胞的形态结构更清晰,能更好地显示出不同种类的细胞及细胞的不同成分的色彩特征,可以更快、更有效地分辨各类细胞的细胞核、细胞浆和被吞噬的外来异物。结论采用瑞氏-姬姆萨染色法制作的吞噬作用实验标本在镜下形态结构清晰、色彩特征明显,易于分辨细胞成分和外来异物的种类,便于学生观察和计数,有助于学生对吞噬细胞吞噬作用的理解和掌握。     

3种方法检验结核分枝杆菌的比较

目的比较直接涂片抗酸染色法、罗氏培养基培养法、环介导等温扩增法(lamp)检测痰结核分枝杆菌的准确度和特异性,及在临床诊断中的应用价值。方法用3种方法对送检的845个痰标本进行检验,并对检验结果进行分析。结果 845个痰标本中,涂片阳性81个,阳性率9.6%;培养阳性136个,阳性率16.1%;lamp阳性124个,阳性率14.7%。结论培养法是检测结核分支杆菌的金方法,检出率高,其次是环介导等温扩增法,其灵敏度较高与培养法检测结果相比差异无统计学意义(P>0.05);涂片法检出率较低,与另外两种方法相比差异有统计学意义(P<0.05)。     

人芽囊原虫标本染色方法的改进

人芽囊原虫标本经肖氏液固定后,用改良的哈氏苏木素染色。结果表明,标本内部结构清晰,中心体明显,与传统的铁苏木素染色方法相比,染色效果好,制片时间短,操作步骤简单。