一种用于制作骨骼组织切片的新脱钙液

在日常病理工作中,对某些骨骼组织需作薄切片观察其细微结构,这就要对骨骼进行脱钙处理.通常使用的脱钙液有以硝酸、盐酸、甲酸为主配制的脱钙液、还有以铬酸和螯合剂配制的脱钙液.     

骨切片固定液和脱钙剂的筛选及效果观察

在研究某些骨病及骨组织内的各种成分时,尽量减少对骨组织内成分的损伤,选择合适的固定液、固定时间和脱钙液,对骨组织制片非常重要。我们查阅了国内外70年代以来的有关文献,找出固定方法40余种,脱钙方法     

一种简便快捷的骨及钙化组织脱钙液

脱钙是骨组织制片染色技术的关键步骤.在实践工作中以硝酸、盐酸为主配制的脱钙液,脱钙迅速,但对组织破坏程度大,影响以后染色.螯合剂是一种缓慢的脱钙剂,脱钙后染色分化仍佳,酶也不受破坏,但脱钙需花费2周至几个月时间,不能满足病理科日常工作的需求[1].现介绍一种混合脱钙液,它既对组织损害小,又简便快捷.它可促进脱钙并加快脱钙速度,同时又防止纤维性组织过度膨胀,减少对组织的破坏及对染色的影响.经此脱钙液制成的组织切片薄厚均匀,平坦完整,组织形态和细胞清晰,着色鲜艳.     

牙和骨切片的粘片剂防脱片效果及应用方法探讨

牙和骨切片HE尤其是免疫组织化学染色需多次更换试剂、改变温度、反复水洗,经常造成切片易皱褶、卷曲、与载玻片分离（脱片）。目前尚无理想的防止牙和骨切片染色脱片的方法,亦未见防止牙和骨切片染色脱片的报道。以下是我们探讨牙和骨切片的氨醛酸乙基硅（APES）、多聚-L-赖氨酸（PLLS）两种粘片剂的防脱片效果和应用方法。     

一种复合固定脱钙液在临床病理制片中的应用

临床病理检查经常遇到骨及钙化组织标本,常规处理方法是先固定,待充分固定后进行脱钙处理,然后进入常规制片程序,到完成病理诊断需要较长的时间。如何在较短时间内完成固定、脱钙处理,其速度快、效果好,同时制成的切片又和常规处理的不含骨组织的切片一样,不影响对病变组织     

四种脱钙液对骨和骨髓组织及结构影响的比较

背景：由于骨和骨髓组织结构非常特殊,在常规的病理制片过程中,要同时显示坚韧质硬的骨组织和含有多种幼稚娇嫩造血细胞的骨髓组织是非常困难的。目的：选择一种既能完全除去骨组织中的钙质,又能保护骨髓组织及细胞结构不受破坏的最佳脱钙液。方法：将含骨髓组织的犬长骨组织随机分4组,同等条件下,14%硝酸甲醛生理盐水溶液;14%硝酸水溶液;20%甲盐酸甲醛生理盐水溶液和20%甲盐酸水溶液4种不同脱钙液脱钙,记录脱钙时间,常规脱水、切片、苏木精-伊红染色,镜下观察,对比4种脱钙液对骨和骨髓组织常规病理切片质量和其苏木精-伊红染色效果。结果与结论：14%硝酸甲醛生理盐水溶液组脱钙能力最强,脱钙最均匀,耗时最短,既能有效完全除去骨皮质中的钙质,又能保护骨髓组织及细胞的形态完好,切片质量和苏木精-伊红染色的效果最佳;14%硝酸水溶液组脱钙液,使骨组织疏松发泡,组织变黄,对组织的损伤大,脱钙不均匀,组织切片不完整,制片质量和苏木精-伊红染色的效果最差;20%甲盐酸甲醛生理盐水溶液组对组织的脱钙能力、损伤程度、制片质量和苏木精-伊红染色效果都比14%硝酸甲醛生理盐水溶液组稍差;20%甲盐酸水溶液组对骨和骨髓组织的损伤小,脱钙效果好,切片质量和苏木精-伊红染色的效果佳,介于20%甲盐酸甲醛生理盐水溶液组和14%硝酸水溶液组之间。结果表明,4种脱钙液对骨和骨髓组织都具有良好的脱钙能力,制片质量及苏木精-伊红染色效果比较,14%硝酸甲醛生理盐水溶液> 20%甲盐酸甲醛生理盐水溶液> 20%甲盐酸水溶液> 14%硝酸水溶液;14%硝酸甲醛生理盐水溶液最适用于临床常规病理制片中骨和骨髓组织的脱钙。中国组织工程研究杂志出版内容重点：组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松;组织工程     

HAS快速脱钙液的研制和应用

水杨酸溶液具有松解胶原、溶解脂肪的作用［１］，临床上常用于疣和鸡眼病的治疗。由于骨组织中的主要成分是无机盐和胶原，故我们试验将水杨酸（ｓａｌｉｃｙｌｉｃａｃｉｄ）与盐酸（ｈｙｄｒｏｃｈｌｏｒｉｃａｃｉｄ）、醋酸（ａｃｅｌｉｃａｃｉｄ）制成快速脱钙液（简称ＨＡＳ液），用于钙化和骨组织标本的快速脱钙，结果显示其具有脱钙作用迅速，组织结构保存良好，切片染色清晰的优点。我们对ＨＡＳ液的研制及应用进行总结，现介绍如下。一、材料与方法１．ＨＡＳ液的配制：取３６％～３８％的盐酸８ｍｌ，９９％醋酸５ｍｌ，水杨…     

超声处理在含骨或沙砾体病理标本脱钙中的应用

病理制片是确诊含有骨化组织及沙砾体组织疾病的主要技术手段之一。常规方法脱钙需4～10h,超声处理可使脱钙时间大大缩短,只需25～60min。我们以对脑膜瘤组织进行脱钙为例,对应用超声处理进行脱钙方法概述如下。[第一段]     

脱钙液在微波辐射下对骨组织作用效果及免疫组织化学染色的影响

骨组织脱钙常规多采用酸性液体（如硝酸、盐酸、甲酸等）进行脱钙，一般需较长时间，组织细胞结构很难保持完整清晰，有些甚至破坏组织结构很严重，从而影响病理学的诊断和研究。骨组织中的蛋白抗原得不到很好的保存，有时甚至丢失。为此我们对９例骨组织用乙二胺四乙酸二...     

Assessment of different decalcifying protocols on Osteopontin and Osteocalcin immunostaining in whole bone specimens of arthritis rat model by confocal immunofluorescence

Confocal immunofluorescence is a valuable technique for the detection of relevant molecules in the pathogenesis of arthritis in rat models; however, it requires efficient processing of tissues including bone decalcification. The decalcification process must ensure the complete removal of calcium and also a proper preservation of cellular structures and, specially, the antigenicity of the tissue to allow the immunodetection of the molecules of interest. In the present study, we evaluated the effect of four different decalcifying solutions: the Morse´s solution, 10% EDTA (pH 7.4), 7% HCl/2% EDTA and 5% Nitric acid, as well as four different treatments of the tissues (including microwave irradiation) in the processes of decalcification for large pieces of adult rat bones (hind paw, fore paw, knee and column). We assessed the time of decalcification, the easiness of slicing, the morphological preservation and finally, the antigenicity of two different bone proteins (Osteopontin (OPN) and Osteocalcin (OC)) measured by its immunofluorescence intensity under controlled confocal microscopy conditions. Our results showed that the specimen size and the presence of skin are critical factors for the rate of decalcification, and no significant benefit was found if microwave irradiation is applied to the tissue. The comprehensive statistical analysis showed that the optimal solution for the detection of OPN and OC by confocal immunofluorescence is the 5% Nitric Acid, and followed by 10% EDTA (pH 7.4), Ana Morse solution and 7% HCl/2% EDTA.     

Antibacterial activity of sucralfate versus aluminum chloride in simulated gastric fluid

Studies have previously demonstrated that sucralfate possesses intrinsic antibacterial activity. This study was designed to indirectly assess whether aluminum is the active antibacterial component of sucralfate and to further evaluate factors that may influence this agent's antibacterial activity. Utilizing an in vitro model, the antibacterial activity of sucralfate, an equivalent quantity of aluminum in the form of aluminum chloride, and a control were compared. In addition, the influences of bacterial species (Enterobacter cloacae andPseudomonas aeruginosa), time (0–24 h) and environmental pH (3, 5, 7) on the agents' antibacterial activities were evaluated. Equivalent quantities of aluminum, as either sucralfate or aluminum chloride, were added to two of three flasks containing approximately 10⁵ cfu/ml of bacteria in pH-adjusted simulated gastric fluid. The third flask served as a control. Samples were obtained over 24 h, diluted and subcultured onto agar plates. The experiments demonstrated that bacterial growth was influenced by pH, time and treatment (aluminum chloride or sucralfate). Regardless of pH or bacterial species, bacterial death occurred within 20 min following the addition of aluminum chloride. In contrast, bacterial death following the addition of sucralfate was more variable and appeared to be pH dependent. In conclusion, sucralfate and aluminum chloride both possess antibacterial activity, even at pH values that normally support bacterial growth in gastric fluid. Although differences in the antibacterial activity of the two agents may in part be related to drug-induced changes in pH, these differences also support data suggesting that aluminum release from sucralfate is incomplete and is dependent on pH.     

试剂对脱钙骨基质成骨能力影响的实验研究

目的 :研究异体骨基质制备过程中常用的脱脂和脱钙试剂溶液对其成骨能力的影响。方法 :8周鼠龄的Wistar雌鼠作为供体 ,切取胫股骨 ,碾磨成 12 5～ 80 0微米的颗粒 ,分成四组 ,分别用丙酮 -盐酸、丙酮 -乙二胺四乙酸 (EDTA)、三氯甲烷 -甲醇 -EDTA或三氯甲烷 -甲醇 -盐酸 (控制组 )脱脂和脱钙。冷冻干燥处理后 ,每 3 0mg装入一胶囊中。 60只 6周鼠龄的Wistar雌鼠作为受体 ,植入腰背肌中 ,每只鼠接受同一组植入材料 2枚。术后 6周取标本测定重量、干燥后的重量、钙含量、碱性磷酸酶含量和组织形态学定量测定。结果 :丙酮 -盐酸组的干燥重量 (P <0 .0 1)和钙含量 (P <0 .0 5 )明显高于控制组。结论 :用丙酮清除脂质的异体骨基质成骨能力优于用三氯甲烷 -甲醇混合液。EDTA或盐酸脱钙对其成骨能力无明显影响。     

Decalcification by ultrasound and acid in a histological routine laboratory

The ultrasonic and acid treatment of biopsy samples is a rapid method for decalcifying bones and yields good preparations. The combination of the sodium sulphate technique and washing by ultrasound requires 24 h. The method described is feasible in routine laboratories without the necessity of high technical expense.     

Immunophenotypic analysis of acute lymphoblastic leukemia using routinely processed bone marrow specimens

Monoclonal antibodies have been recently developed that react with antigens expressed on T and B lymphocytes in routinely processed, paraffin-embedded lymphoid tissues. In this study, we assessed bone marrow clot and/or core biopsy sections of 19 cases of acute lymphoblastic leukemia (ALL) using routinely decalcified, B5- or formalin-fixed, paraffin-embedded sections and a panel of monoclonal antibodies, including LN1, LN2, L26, Leu-22, UCHL-1, and LCA. Each case had been previously phenotyped using freshly obtained aspirate material and a standard immunophenotypic protocol. Our results demonstrate the utility of the LN2 antibody in differentiating between precursor B-cell (pre-B) and precursor T-cell ALL. The LN2 antibody stained 11 of 12 cases of pre-B ALL and did not react with any of the seven T-cell ALLs. The other antibodies tested were less helpful. The Leu-22 antibody stained both pre-B and T-cell ALLs, while the results with UCHL-1 revealed peculiar nuclear staining of pre-B and T-cell ALLs; this we attributed to processing artifact. The L26 antibody reacted with only one case of pre-B ALL (also CD20 antigen positive), while the LN1 antibody did not react with any pre-B ALLs. Neither L26 nor LN1 stained any cases of T-cell ALL. The LCA antibody stained in only four (21%) of 19 cases, two pre-B and two T-cell ALLs. The results also suggest that this panel of antibodies may be useful in differentiating ALL from mature B-cell and T-cell lymphomas involving the bone marrow.     

Infra-red photography of bone and joint specimens

Summary Photographing with the aid of short-wave infra-red light has been used for improved photographic demarcation of hyaline cartilage from connective tissue and fibrous cartilage in specimens from children. This improved effect seems to be the result of a particularly high permeability to infra-red light rays in the hyaline cartilage of children.

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Zusammenfassung Das Photographieren mit kurzwelligem infrarotem Licht wurde zur verbesserten photographischen Begrenzung des hyalinen Knorpels gegenüber dem Bindegewebe und dem fibrösen Knorpel bei Präparaten von Kindern benutzt. Der Effekt scheint auf eine speziell hohe Durchlässigkeit beim hyalinen Knorpel des Kindes für die infrarote Strahlung zurückzuführen zu sein.