三色法显示小肠潘氏、杯状、上皮细胞

本文根据潘氏、杯状细胞都具有合成分泌多糖类物质的特性,先将小肠切片作PAS法染色,再进行胞质、服核复染。这样制作的切片标本,能直观地显示出小肠潘氏、杯状、上皮细胞的分布情况以及它们各自的形态     

室间质控中AB-PAS染色的异常着色原因分析

目的 分析室间质控工作中AB-PAS染色异常着色产生的原因。方法 选取有局部肠上皮化生的完整胃壁组织的连续切片,分别进行PAS和AB单一染色,以及以下3种不同顺序的AB、PAS和Mayer苏木精（Mayer）联合染色：ABPAS-Mayer、PAS-AB-Mayer或PAS-Mayer-AB联合染色。结果 单一PAS染色时,胃黏膜上皮内的中性黏液和化生肠上皮杯状细胞内的混合性黏液均显示玫瑰红色;单一AB染色时,仅化生肠上皮杯状细胞内的混合性黏液显示湖蓝色;AB-PASMayer联合染色时,胃小凹上皮呈玫瑰红色,化生肠上皮呈紫蓝色,差异明显且细胞核着色清晰;PAS-AB-Mayer联合染色时,胃小凹上皮和化生肠上皮内杯状细胞均呈蓝紫色,无法区别,但细胞核着色正常;PAS-Mayer-AB联合染色时,胃小凹上皮和化生肠上皮内杯状细胞均呈蓝紫色,无法区别,而且细胞核着色不佳。结论 行AB和PAS联合染色时必须严格按照先行AB染色后行PAS染色,最后行苏木素Mayer染色的染色步骤,不能将染色顺序随意打乱,否则会出现中性、酸性和混合性黏液物质混染,或者细胞核不着色等异常染色结果,进而干扰病理诊...     

胡子鲶粘液细胞类型及其在消化道中的分布

利用阿新兰过碘酸雪夫试剂染色方法 ,对胡子鲶粘液细胞类型以及在消化道中的分布进行了研究。根据AB PAS染色结果 ,将粘液细胞分为四型 :Ⅰ型 ,AB PAS染色呈红色 ,含有PAS阳性的中性粘多糖 ;Ⅱ型 ,AB PAS染色呈蓝色 ,含有AB阳性的酸性粘多糖 ;Ⅲ型 ,AB PAS染色呈紫红色 ,主要含有PAS阳性的中性粘多糖 ,同时含有少量AB阳性的酸性粘多糖 ;Ⅳ型 ,AB PAS染色呈蓝紫色 ,主要含有AB阳性的酸性粘多糖 ,同时含有少量PAS阳性的中性粘多糖。同时发现粘液细胞在消化道的各段均有分布 ,但是不同部位的粘液细胞数和类型均有差异。     

一种同时显示十二指肠内、外分泌细胞的染色方法

通常组织学教学用十二指肠切片标本多采用传统的H·E染色法制作,使用该种切片标本,无法看到内分泌细胞,仅能观察外分泌细胞和上皮细胞,且所见杯状细胞仅呈无色、空泡状。而同时在切片上显示十二指肠内、外分泌细胞的染色方法,目前国内尚无报道。我们经过反复试验,摸索出一种新染色方法,可在同一张切片上直接显示出十二指肠亲银细胞、杯状细胞和十二指肠腺腺细胞,所显示的各种分泌细胞颜色各异,对比分明,易于观察、辨别。方法如下:     

人结肠切片三种特殊染色的比较

目的比较人结肠组织切片阿利辛蓝AB染色(Alcian Blue)、过碘酸雪夫氏PAS染色(Periodic Acid Schiff)及洋红三种染色方法及镜下特点。方法取人结肠,Carnoy氏固定液固定,对结肠组织进行阿利辛蓝AB染色、PAS染色及洋红染色,光学显微镜下观察染色效果。结果三种染色都能清晰显示结肠组织的结构,尤其对结肠的粘膜层进行了观察。阿利辛蓝染色使结肠上皮吸收细胞的细胞核染为红色,杯状细胞中的黏液染为蓝紫色;洋红染色使结肠上皮吸收细胞的细胞核染为蓝紫色,杯状细胞中的黏液染为红色。结论阿利辛蓝和洋红染色均能清楚显示杯状细胞中的黏液,且染色后的细胞质和细胞核成为互补色,结构清晰,对比明显。     

中华鳖肾脏的组织化学特性研究

利用光镜组织化学反应对中华鳖肾单位的结构和组织化学特性进行了详细的观察和分析。结果表明,中华鳖肾脏为分叶形的实质器官,肾小叶由被膜和实质组成,实质无髓质和皮质之分,但可以区分为外侧区和内侧区。外侧区嗜酸性,主要分布有近端小管和集合管。内侧区呈弱嗜酸性,肾小体、颈段、中间段和远端小管主要分布在内侧区。肾小球PAS反应呈阳性,但其琥珀酸脱氢酶(SDH)弱阳性,碱性磷酸酶(ALPase)、Na+/K+-ATPase和阿利新兰(AB)反应为阴性。足细胞酸性磷酸酶(ACPase)反应呈阳性。近端小管刷状缘嗜伊红,PAS反应以及ALPase、ACPase和Na+/K+-ATPase酶反应呈阳性,而SDH弱阳性。中间段、远端小管、集合管弱嗜酸性,SDH阳性。中间段Na+/K+-ATPase弱阳性。远端小管细胞侧面呈PAS阳性,腔面显示AB阳性。集合管胞质含有许多ACPase阳性颗粒,腔面呈PAS强阳性,AB阳性。甲苯胺兰(TB)染色可见集合管腔面有阳性颗粒,肾小管上皮含有亮、暗两种细胞。上述组化反应和分布结果表明,鳖的肾小管细胞类型较多,近端小管在原尿的重吸收中起主要作用,远端小管和集合管具有分泌黏液作用。中华鳖肾单位的结构与组化特性不仅与哺乳类和鸟类有一定差异,也与其他爬行动物不完全相同。       

鲇肠道黏液细胞的类型、分布、发育及分泌方式研究

利用阿利新蓝-过碘酸雪夫氏试剂(AB-PAS,AB染液pH2．6),对鲇(Silurus asotus)肠道黏液细胞的类型与分布进行了研究,同时采用扫描和透射电镜,研究了黏液细胞的发育与分泌方式。结果显示:鲇肠道黏液细胞染色后呈深蓝色、淡蓝色和浅蓝色,主要含有酸性黏多糖:形态上进一步可将其分为囊状、梨状和杯状三种形态。肠道中黏液细胞的分布密度以中肠最高。其次是后肠,前肠最低;肠道黏液细胞由肠黏膜基底层细胞衍生后,不断向肠腔表层推移而发育成熟;发育过程中黏液细胞内富含大小不等的黏原颗粒,其分泌方式为顶浆分泌。     

豚鼠小肠EC细胞的免疫细胞化学和银染法的比较研究

本实验利用同一切片标本,在光镜水平上,对豚鼠小肠ＥＣ细胞进行了免疫细胞化学和不同银染法的比较研究。实验结果显示,全部Ｍａｓｓｏｎ－Ｆｏｎｔａｎａ亲银染色阳性细胞均呈ＰＡＰ染色阳性;另一方面,小部分（１５％）Ｇｒｉｍｅｌｉｕｓ嗜银染色阳性细胞却表现为亲银染色和ＰＡＰ染色双阴性。以上结果表明,在显示ＥＣ细胞上,亲银染色具有很高的特异性,因此仍不失为一种经济和实用的方法;嗜银染色显示ＥＣ细胞,尽管其敏感性稍高于亲银染色法,但不具有免疫细胞化学与亲银染色技术那样明显的特异性。     

以偶氮偶联(azo-conpling)及粘蛋白染色的对比作脑垂体前叶诸型细胞的组织化学鉴别

根据 Pearse 氏偶联四氮盐反应(coupled tetrazo-nium)染脑下垂体,其嗜酸嗜硷两种细胞皆可染色,仅前者着色较深,故不宜于组织学鉴别之用。作者鉴于嗜硷细胞的向性腺及向甲状腺激素皆系粘蛋白(mucopro-tein),故染粘蛋白的高碘酸白复红法(periodic acid-leukofuchsin)可作力重氮蓝B(diazo llue B)或重氮蓝B和H酸(H acid)联用的对此染色。作者也鉴于用重氮蓝B可加强嗜酸性细胞对于某些染料的亲和力(如固绿FCF)而描述了应用固绿 FCF(fastgreen FCF)的偶氮偶联染色法。     

消化道组织块黏液细胞的组织化学染色

目的：探索中华蟾蜍、黑斑蛙消化道组织块黏液细胞的组织化学染色。方法：利用阿辛蓝-过碘酸Schiff（AB-PAS）反应,对消化道组织块黏液细胞进行染色和观察。结果：对消化道壁较薄的组织块（食管、小肠、大肠）,采用3％乙酸3min、1％阿尔新蓝30min、3％乙酸3min、3％过碘酸氧化10min、Schiff反应20min染色,即可达到良好染色效果;对消化道壁较厚的组织块（胃）,则应采用3％乙酸9min、1％阿尔新蓝70min、3％乙酸9min、3％过碘酸氧化30min、Schiff反应60min染色,将能达到良好染色效果。结论：经过染色可将黏液细胞分为四个类型：Ⅰ型红色,PAS染色阳性,AB染色阴性;Ⅱ型蓝色,PAS染色阴性,AB染色阳性;Ⅲ型紫红色,PAS染色强阳性,AB染色弱阳性;Ⅳ型蓝紫色,PAS染色弱阳性,AB染色强阳性。     

Histochemical distribution of four types of enzymes and mucous cells in the gastrointestinal tract of reared half‐smooth tongue sole Cynoglossus semilaevis

The histochemical distribution of acid phosphatase (ACP), alkaline phosphatase (ALP), non‐specific esterase (NSE), peroxidase (POD) and mucous‐cell types was evaluated in the gastrointestinal tract of the half‐smooth tongue sole Cynoglossus semilaevis. The enzymes were detected in the entire stretch of the gastrointestinal tract. ACP activity was found in the supranuclear region of enterocytes and the lamina propria of the intestine, as well as the cytoplasm of epithelial cells of the stomach. The staining intensity of ACP in the anterior and posterior intestines was stronger than in the stomach. ALP activity was detected in the striated border of enterocytes and muscularis of the whole intestine, lamina propria and supranuclear cytoplasm of the enterocytes in the anterior intestine, as well as in the blood vessels of the stomach. The staining intensity for ALP in the anterior intestine was stronger than in the posterior segment and the latter was stronger than in the stomach. NSE activity was detected in the cytoplasm of the epithelial cells in the entire gastrointestinal tract, with the anterior intestine showing stronger intensity than the stomach. POD activity was located in the blood cells of the lamina propria of the gastrointestinal tract and the levels in the stomach were similar to the anterior and posterior intestines. Alcian blue (pH 2·5) periodic acid Schiff (AB‐PAS) histochemical results revealed three types of mucous cells in the gastrointestinal tract. Type I cells (PAS+AB‐) were observed among the gastric mucosa columnar cells in the stomach and enterocytes in the basal region of the villi and in the middle and top regions of the intestinal villi. Type II cells (PAS‐AB+) and type III cells (PAS+AB+) were not detected in the stomach but were distributed ubiquitously among enterocytes in the middle and top regions of the intestinal villi.     

Histochemical and Biometric Study of the Gastrointestinal System of Hyla Orientalis (Bedriaga, 1890) (Anura,Hylidae)

This study was carried out to assess the localization of hyaluronic acid (HA) and the distribution of glycoproteins in the gastrointestinal system of adult Hyla orientalis. Histochemical analysis of the gastrointestinal system in H. orientalis showed that mucous content included glycogene and/or oxidable dioles [periodic acid/Schiff (PAS)+], neutral or acid-rich (PAS/AB pH 2.5+), sialic acid residues (KOH/PAS+) and acid sulphate [Aldehyde fuchsin (AF)+] glycoproteins. However the mucus content was not the same in stomach, small and large intestine. The mucus content of stomach included only glycogene and/or oxidable dioles and sialic acid residues. Besides these histochemical methods, the localization of HA was detected using biotinylated hyaluronic acid binding protein labeled with streptavidin-fluorescein isothiocyanate (FITC). In the extracellular matrix of the submucosa, the reaction for HA was evident. Since HA was located in submucosa beneath the epithelial layer of gastrointestinal system, it has a significant role in hydric balance, and essential to provide the gastrointestinal system integrity and functionality. According to biometric results, there were statistical differences between small and large intestine in terms of the amount of material stained positive with PAS/AB, PAS, KOH/PAS and AF/AB. Additionally, number of goblet cells in the small and large intestine was significantly different.Key words: Gastrointestinal system, goblet cell, glycoproteins, hyaluronic acid, amphibian, Hyla.     

Histochemical study on the intestine goblet cells in cichlid and poecilid species (Teleostei)

Histochemical properties of intestine goblet cells in firemouth cichlid, zebra mbuna, freshwater angelfish and platyfish are described. Goblet cells occurred regularly in the epithelial cell layer throughout the entire intestine, they were strongly coloured by alcian blue at pH 2.5. This colour got gradually weaker when the pH was reduced, but still after alcian blue at pH 0.2 these cells displayed a distinct blue colour. When the goblet cells were treated with periodic acid-Schiff (PAS), they displayed a strong purple-magenta colour. The findings that a number of goblet cells displayed various colours between blue and purple-magenta when acidic alcian blue was followed by PAS, and between blue and red-brown when acidic alcian blue was followed by neutral red, may reflect different ages or stages of development and differentiation for these cells. However, such results may also suggest a true cellular heterogeneity in the present population of goblet cells, reflecting that the intestine mucus layer has a number of roles in teleosts like lubrication, protection, immunological defence, digestion and absorption.In the ferritin injected specimens of firemouth cichlid and platyfish, a number of macrophage-like cells in intestine wall displayed Prussian blue precipitations in tissue treated with acid ferrocyanide, suggesting that these cells play a cleansing role in the intestinal wall. No ferritin uptake was seen in the intestine goblet cells and eosinophilic granule cells.     

Histochemical features of the Muscovy duck small intestine during development

We demonstrated for the first time the distribution and morphology of argyrophil and of goblet cells in the mucosa of the small intestine of the Muscovy duck during development using the Grimelius silver staining and alcian blue/periodic acid-Schiff (AB/PAS) staining technique. The argyrophil cells distribution was variable over the length of the small intestine from embryonic day 24 (24E) to post-hatching day 13 (13d). In the villi most argyrophil cells belonged to the open-type, while in the crypts they belonged to the closed-type. In the duodenum the density of argyrophil cells was highest at hatching, while in the jejunum and in the ileum the highest density value was at hatching and 13d. AB/PAS-positive goblet cells appeared on the villi and crypts of the duodenum and jejunum at 30E, and in the ileum at hatching. The density of AB/PAS-positive cells was the highest in the three segments at hatching. The AB-positive cells, compared with the PAS-positive cells, predominated in villi and crypts of the three segments, moreover the rate of AB-positive cells to PAS-positive cells significantly decreased from 30E to 9d. An increase in argyrophil and goblet cells number during the later incubation and at hatching, could indicate the small intestine in that period is being prepared to face a new diet.     

Autophagy proteins control goblet cell function by potentiating reactive oxygen species production

Delivery of granule contents to epithelial surfaces by secretory cells is a critical physiologic process. In the intestine, goblet cells secrete mucus that is required for homeostasis. Autophagy proteins are required for secretion in some cases, though the mechanism and cell biological basis for this requirement remain unknown. We found that in colonic goblet cells, proteins involved in initiation and elongation of autophagosomes were required for efficient mucus secretion. The autophagy protein LC3 localized to intracellular multi‐vesicular vacuoles that were consistent with a fusion of autophagosomes and endosomes. Using cultured intestinal epithelial cells, we found that NADPH oxidases localized to and enhanced the formation of these LC3‐positive vacuoles. Both autophagy proteins and endosome formation were required for maximal production of reactive oxygen species (ROS) derived from NADPH oxidases. Importantly, generation of ROS was critical to control mucin granule accumulation in colonic goblet cells. Thus, autophagy proteins can control secretory function through ROS, which is in part generated by LC3‐positive vacuole‐associated NADPH oxidases. These findings provide a novel mechanism by which autophagy proteins can control secretion.     

Comparative histochemistry of the mucoproteic cells of the hypophysis from Rhamdia hilarii, Hypostomus punctatus, Prochilodus scrofa and Cyprinus carpio (Teleostei). Immunohistochemical identification of the gonadotropic cells

Adenohypophyseal cells showing positive histochemical reactions for mucosubstances were classified as type I-IV in Hypostomus (Plecostomus) punctatus (Loricariidae), Rhamdia hilarii (Pimelolidae), Prochilodus scrofa (Prochilodontidae) and Cyprinus carpio (Cyprinidae) according to cell shape, size, cytological characteristics and adenohypophyseal distribution. Cell types I and II are common to the four species, with each cell type showing very similar cytological and histochemical characteristics, in spite of different adenohypophyseal distribution of cell type II, according to the teleost species. Type I cells are globular basophils located in the proximal pars ditalis and are positive to PAS and Alcian blue pH 2.5 (AB) reactions, showing cytoplasmic vacuoles and changes in granule concentration in the mature phase of the gonadal cycle. The smaller type II cells are fusiform or oval basophils exhibiting a strong AB reaction but also reacting to PAS. Type III cells are located in the pars intermedia showing PAS-positive reaction. Considering different teleost species, these cells exhibit some variations specially in relation to cell size and shape which are not detected in mature male C. carpio. Otherwise cell type IV is only present in the rostral pars distalis of P. scrofa. They are weakly basophilic and negative to PAS, reacting strongly to AB. Only cell type I showed unequivocally positive immunohistochemical results with anti-salmon gonadotropin.