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A modified mallory-cason staining procedure for large cryosections. 一种用于大切片的改良马氏染色方法。
Pub Date : 1990-01-01 DOI: 10.3109/10520299009105606
M B van Leeuwen, A J Deddens, P O Gerrits, B Hillen

The classic Mallory-Cason staining procedure has been modified for application to sections "on tape" obtained from large deep frozen tissue specimens. These 20 microns cryosections are collected on tape from a large heavy duty cryomicrotome. The stained sections provide anatomical details that are not revealed by other techniques. The merit of this procedure is found in the support of modern medical modalities, both for research and educational purposes.

经典的Mallory-Cason染色程序已被修改,适用于从大型深度冷冻组织标本中获得的“磁带上”切片。这些20微米的冷冻切片是从一个大型重型冷冻切片机上收集的。染色切片提供了其他技术无法揭示的解剖细节。这一程序的优点在于支持现代医学模式,既用于研究也用于教育目的。
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引用次数: 27
New stains for blood and bone marrow cells. 新的血液和骨髓细胞染色剂。
Pub Date : 1990-01-01 DOI: 10.3109/10520299009105612
L Kass

Traditionally, blood and bone marrow cells have been identified based on their characteristic shapes and colors when stained with one of several panoptic stains including Wright's or Giemsa's. As questions arose regarding the origin of normal and leukemic cells, cytochemical stains were developed. These stains help identify cells on the basis of a distinctive metabolite or enzyme. As part of an ongoing tradition in which textile dyes are used for biological staining, several new stains have been applied to hematologic staining. These include C.I. basic blue 41, basic blue 141, basic blue 93, and an asymmetrical polymethine dye. As additional cell-selective stains are developed, we can anticipate further improvements in our ability to identify normal and malignant hematopoietic cells.

传统上,血液和骨髓细胞是根据它们的特征形状和颜色来识别的,这些细胞是用几种全光染色剂之一染色的,包括赖特染色剂或吉姆萨染色剂。随着对正常细胞和白血病细胞来源的疑问的出现,细胞化学染色被开发出来。这些染色有助于根据一种独特的代谢物或酶来识别细胞。作为正在进行的传统的一部分,纺织染料用于生物染色,一些新的染色已应用于血液学染色。这些染料包括C.I.碱性蓝41、碱性蓝141、碱性蓝93和一种不对称的聚甲基染料。随着其他细胞选择性染色的发展,我们可以预期我们识别正常和恶性造血细胞的能力将进一步提高。
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引用次数: 1
A method to determine the end point of decalcification of hard tissue and bone. 一种测定硬组织和骨脱钙终点的方法。
Pub Date : 1990-01-01 DOI: 10.3109/10520299009108060
S Müller, J Pleul, M Götz, J Jähnig, H Schädlich

A method for decalcification end point determination of mineralized tissue is described. The calcium content of the decalcification solution was determined colorimetrically with a "continuous automatic analyzer" with a high degree of accuracy. The end point method used has been tested on two decalcification methods, 5% nitric acid with or without ultrasonic treatment. The results suggest it is possible to quantitate the decalcification process.

描述了一种矿化组织脱钙终点测定方法。脱钙溶液的钙含量用高精度的“连续自动分析仪”比色法测定。用终点法在5%硝酸加或不加超声处理两种脱钙方法上进行了试验。结果表明,对脱钙过程进行定量分析是可行的。
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引用次数: 3
Immunohistochemical detection of DNA replication in mouse uterine cells by bromodeoxyuridine labeling of wax- and resin-embedded tissue sections. 用溴脱氧尿苷标记蜡包埋和树脂包埋组织切片的免疫组化方法检测小鼠子宫细胞DNA复制。
Pub Date : 1990-01-01 DOI: 10.3109/10520299009139927
M Hanazono, A Yoshiki, K Ota, J Kitoh, M Kusakabe

To apply the bromodeoxyuridine (BrdU) labeling method using a monoclonal antibody to the study of cell proliferation in the mouse uterus, methods of fixation and embedding of tissues and of immunofluorescent staining were compared in terms of the rate of detection of labeled cells and specificity and stability of fluorescence obtained. BrdU was administered intravenously 2 hr before death and uterine blocks were embedded in polyester wax and Technovit resin after fixation in formalin and periodate-lysine-paraformaldehyde, respectively. The indirect method with anti-BrdU and fluorescein isothiocyanate (FITC) conjugated antimouse IgG antisera and the direct method with FITC conjugated anti-BrdU antibody were applied to both wax- and resin-embedded sections. Labeled and total cells were counted in luminal and glandular epithelia and stomata adjoining them. Counterstaining with hematoxylin for counting total cells produced intense fluorescence over the whole of resin sections and made counting of labeled cells impossible. On wax sections, on the other hand, the results were satisfactory, although the number of labeled cells detected was decreased slightly. In wax sections fluorescence due to nuclear incorporation of BrdU in the indirect method could be easily distinguished from the cytoplasmic or extracellular emission seen in some cells by its location and characteristic color. In resin sections, however, more careful observation was needed since the second antibody used in the indirect method cross-reacted with IgG in eosinophils and produced cytoplasmic fluorescence of the same color. By the indirect method greater numbers of labeled cells were detected in wax sections than in resin sections. The difference was distinct in tissues with extensive cell proliferation. By the direct method the fluorescence obtained was weaker and apt to fade more quickly than that obtained by the indirect method; use of the direct method reduced the number of labeled cells detected in both wax- and resin-embedded sections.

为了将溴脱氧尿苷(BrdU)单克隆抗体标记法应用于小鼠子宫细胞增殖的研究,比较了组织固定包埋法和免疫荧光染色法对标记细胞的检出率、荧光特异性和稳定性的影响。死前2小时静脉给予BrdU,分别用福尔马林和高碘酸-赖氨酸-多聚甲醛固定后,将子宫块包埋在聚酯蜡和Technovit树脂中。蜡包埋切片和树脂包埋切片分别采用抗brdu和异硫氰酸荧光素(FITC)偶联抗小鼠IgG抗血清间接法和FITC偶联抗brdu抗体直接法。计数管腔上皮、腺上皮及相邻气孔的标记细胞和总细胞。用苏木精反染色计数总细胞会在整个树脂切片上产生强烈的荧光,使标记细胞计数不可能。另一方面,在蜡切片上,虽然检测到的标记细胞数量略有减少,但结果令人满意。在蜡切片中,由于BrdU的核融合,间接方法中的荧光可以很容易地与某些细胞的细胞质或细胞外发射区分开来,通过其位置和特征颜色。然而,在树脂切片中,由于间接方法中使用的第二抗体与嗜酸性粒细胞中的IgG交叉反应并产生相同颜色的细胞质荧光,因此需要更仔细的观察。通过间接方法,蜡切片比树脂切片检测到更多的标记细胞。在细胞广泛增殖的组织中差异明显。与间接法相比,直接法获得的荧光较弱,容易褪色;使用直接法减少了蜡和树脂包埋切片中检测到的标记细胞的数量。
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引用次数: 17
Localization of healing tracheal wounds using bromodeoxyuridine immunohistochemistry. 溴脱氧尿苷免疫组织化学用于气管伤口愈合的定位。
Pub Date : 1990-01-01 DOI: 10.3109/10520299009105604
E M McDowell, X M Zhang, A M DeSanti

Immunohistochemical demonstration of the thymidine analogue bromodeoxyuridine (BrdU) in regenerating cells was useful in determining the size and location of the wounded areas (epithelial and submucosal) during regeneration of the hamster tracheal epithelium, at times late in the healing process (72-144 hr postinjury) when the wound sites and their boundaries were not recognized with certainty in conventionally stained paraffin sections. Cells distant from the wound sites remained unlabelled. The success of this method resulted from prolonged exposure to BrdU released over several hours from a 25mg tablet implanted subcutaneously at 24 hr postwounding at the time when DNA synthesis and cell proliferation are maximal. This simple technique promises to be useful in determining the size and location of wound sites with application to a wide variety of organs and tissues in studies of repair and healing.

再生细胞中胸腺嘧啶类似物溴脱氧尿嘧啶(BrdU)的免疫组织化学证明有助于确定仓鼠气管上皮再生过程中受伤区域(上皮和粘膜下)的大小和位置,有时在愈合过程后期(损伤后72-144小时),当常规染色石蜡切片无法确定伤口位置及其边界时。远离伤口部位的细胞仍未被标记。该方法的成功是由于在损伤后24小时皮下植入25mg片剂,在几个小时内释放BrdU,这是DNA合成和细胞增殖最大的时间。这项简单的技术有望用于确定伤口部位的大小和位置,并在修复和愈合的研究中广泛应用于各种器官和组织。
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引用次数: 15
Microwave fixation of the lung. 微波固定肺。
Pub Date : 1990-01-01 DOI: 10.3109/10520299009108063
C R Turner, S Zuczek, D J Knudsen, E B Wheeldon

A technique for microwave fixation of inflated rat lung is described. Conventional intratracheal fixation with instillation of fixative into the airways at a constant pressure results in pressure artifacts as well as flushing and disruption of cells and exudates. Microwave fixation fixes these elements in situ without disruption and thus is valuable when evaluating the distribution of inflammatory infiltrates. Exudative pneumonitis was produced in the rat using intratracheal instillations of either endotoxin or silica and comparisons were made between histologic sections fixed using either standard formalin fixation or microwave fixation.

介绍了一种微波固定膨胀大鼠肺的方法。常规气管内固定以恒压向气道内灌注固定剂,导致压力伪影以及细胞和渗出液的冲洗和破坏。微波固定将这些元素固定在原位而不破坏,因此在评估炎症浸润分布时是有价值的。在气管内注入内毒素或二氧化硅引起大鼠的渗出性肺炎,并用标准福尔马林固定或微波固定固定组织学切片进行比较。
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引用次数: 17
Chromatin fluorescence after carmine staining. 胭脂红染色后的染色质荧光。
Pub Date : 1990-01-01 DOI: 10.3109/10520299009105621
J C Stockert, A R Llorente, P Del Castillo, A Gómez

After staining with dilute solutions (0.1 mg/ml in distilled water) of commercial carmine, a strong reddish orange fluorescence was observed in nuclei from cell smears and frozen and paraffin tissue sections. Optimal exciting light was 436 nm (violet-blue) or 450-490 nm (blue). Compact chromatin from interphase nuclei, mitotic and meiotic chromosomes and the kinetoplast of Trypanosoma cruzi showed the highest fluorescence, while the basophilic cytoplasm appeared weakly fluorescent. No emission was observed in cartilage matrix, mast cell granules or goblet cell mucin. This selective method could be valuable in microscopic and cytochemical studies on chromatin because the carmine fluorescence is stable and preparations can be dehydrated and mounted permanently without changes in the fluorescence pattern.

用商业胭脂红稀释溶液(0.1 mg/ml蒸馏水)染色后,在细胞涂片、冷冻和石蜡组织切片的细胞核中观察到强烈的红橙色荧光。最佳激发光为436 nm(紫蓝色)或450-490 nm(蓝色)。克氏锥虫间期核致密染色质、有丝分裂和减数分裂染色体和着丝质体的荧光最强,而嗜碱性细胞质的荧光较弱。软骨基质、肥大细胞颗粒、杯状细胞黏蛋白均未见释放。这种选择性方法在染色质的显微镜和细胞化学研究中是有价值的,因为胭脂红荧光是稳定的,并且制备物可以脱水和永久安装而不会改变荧光模式。
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引用次数: 8
A simple procedure to visualize osmicated storage lipids in semithin epoxy sections of plant tissues. 在植物组织的半薄环氧树脂切片中可视化渗透储存脂的简单程序。
Pub Date : 1990-01-01 DOI: 10.3109/10520299009105608
E C Yeung
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引用次数: 20
Differential staining of the cell cycle of plant cells using safranin and indigo-picrocarmine. 用藏红花红和靛蓝-微胭脂红对植物细胞周期进行鉴别染色。
Pub Date : 1990-01-01 DOI: 10.3109/10520299009108070
D Swain, D N De

A trichrome staining technique using safranin-indigo-picrocarmine (SIPC) can be used to distinguish the various stages of the cell cycle in onion root tip. When the tissue was fixed first in formalin followed by picric acid and stained in SIPC, a clear differentiation of interphase nuclei into four color classes, viz., green, orange, red and brown can be recorded. Replacing crystal violet for safranin produces a similar pattern of differentiation of interphase nuclei into green, light blue, blue and deep blue. Autoradiographic study using 3H-thymidine as a DNA precursor demonstrates the reliability of the SIPC staining technique. All the orange and red nuclei are found to be labelled and therefore are in S phase of the cell cycle. Almost all the green nuclei are unlabelled and may be assigned to G1. The larger brown nuclei which are mostly unlabelled can be considered in G2 phase.

红花-靛蓝-微胭脂碱(SIPC)三色染色技术可用于区分洋葱根尖细胞周期的不同阶段。将组织先用福尔马林再用苦味酸固定,SIPC染色,可见间期细胞核明显分化为绿色、橙色、红色和棕色四种颜色。用水晶紫代替红花红可以产生类似的间期细胞核分化为绿色、浅蓝色、蓝色和深蓝色的模式。使用3h -胸腺嘧啶作为DNA前体的放射自显影研究证明了SIPC染色技术的可靠性。所有橙色和红色的细胞核都被标记,因此处于细胞周期的S期。几乎所有的绿色细胞核都没有标记,可能属于G1。大部分未标记的较大的棕色核可以认为处于G2期。
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引用次数: 2
A solution for removal of resin from epoxy sections. 一种去除环氧树脂的溶液。
Pub Date : 1990-01-01 DOI: 10.3109/10520299009108071
T Iwadare, E Harada, S Yoshino, T Arai

Development of a resin-dissolving solution for use at low alkali concentrations is described. Crown ether dissolved in dimethyl-sulfoxide produces a superbasic alkoxide anion. A five minute treatment resulted in complete resin removal from kidney biopsy specimens embedded in Epon 812. Specimens were well stained by Loeffler's methylene blue. Periodic acid-methenamine silver and Giemsa stains yielded good results. Application of PAS reaction and subsequent hematoxylin counterstaining was practicable for diagnosis.

介绍了一种用于低碱浓度的树脂溶解溶液的研制。冠醚溶解在二甲亚砜中产生超碱性醇氧阴离子。5分钟的治疗导致Epon 812包埋的肾活检标本的树脂完全去除。标本经勒夫勒亚甲基蓝染色。周期酸-甲基苯丙胺银染色和吉姆萨染色结果良好。应用PAS反应和随后的苏木精反染色对诊断是可行的。
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引用次数: 23
期刊
Stain technology
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