介绍一种微波水浴辐射快速石蜡切片法

传统的快速石蜡切片技术主要是利用酒精灯、电炉直接或水浴加热处理组织,以缩短组织的固定、脱水、透明和浸蜡的时间,但这种方法的缺点是组织受热时间和程度难以准确地控制,容易引起组织过脆或处理不足,从而影响诊断。因此,我们尝试利用微波水浴辐射处理组织,经过反复试验和不断改进,摸索出一种制片效果相当理想的微波水浴辐射快速石蜡切片法,在术中病理诊断中取得满意的效果,现介绍如下。     

介绍一种快速冷冻切片方法

快速冷冻切片有利于临床医师在术中获取病理诊断,以便决定进一步的治疗方案。我们在实践中摸索出一种快速、简便、高质量的－２５℃直接骤冷冻切片法,现介绍如下。１　材料与方法１－１　材料　日本产樱花牌恒冷切片机,我院术中冷冻切片标本１５２例。１－２　方法　将取材后的新鲜组织直接放在组织托上,底部、周围及表面都不加ＯＣＴ等包埋剂（穿刺活检、胃镜等微小组织,可预先在组织托上加少许ＯＣＴ等包埋剂,冷冻,再加上微小组织）,迅速置于冷冻台上,于－２５℃恒温下骤冻１～２ｍｉｎ后切片,厚度３～５μｍ,将切片直接贴附于…     

一种改良的石蜡切片显示琥珀酸脱氢酶的方法

目前恒冷箱切片装置虽已简化,价值较前便宜,但仍然属较昂贵的进口仪器,因而大大限制了一些脱氢酶组织化学方法的应用。为此,我们因工作需要摸索出用石蜡切片方法显示琥珀酸脱氢酶(SDH)的方法,现将此法介绍如下: 材料:选用成年杂种雄性小白鼠,取腓肠肌、小肠、睾丸、肝、肾等组织。     

介绍一种冷冻切片快速染色方法

制作优质的冷冻切片,以缩短染色时间、提高染色细胞的核浆清晰度最为重要。我们试将苏木精。伊红按比例配成混合液使整个染色过程在1min内完成,且不必经过盐酸乙醇分化,染色鲜艳而稳定,现介绍如下。1材料和方法1．1染液配制（1）冷冻固定液：85％乙醇85ml,4％甲醛10ml,冰醋酸5ml。（2）苏木精伊红混合染色液：沉淀酸化伊红Y乙醇应用液1份,哈雷苏木精液3份混合供染色用。1．2切片制作取手术新鲜组织置恒冷式切片机载物台上,－20℃冷冻后6μm切片,玻片附贴切片后即刻用冷冻固定液固定10S,水稍洗,滴加苏木精伊红混合染色液至液体覆…     

介绍一种冰冻切片快速染色法

介绍一种冰冻切片快速染色法毕学杰，薛春梅冰冻切片在保证制片质量的前提下，所用时间越短，对临床越有利。我们配制了一种快速冰冻染色液，使染色时间在５ｍｉｎ内即可完成，染色效果和原来慢速染色法相同，现介绍如下。１材料和方法１．１染液配制所用试剂有１０％福尔...     

介绍一种冷冻切片快速免疫组化染色法

对淋巴瘤、癌、恶性黑色素瘤及慢性炎症的冷冻切片 ,仅靠形态学诊断有时非常困难 ,免疫组化能有所帮助。但常规的免疫组化染色需时较长 ,因此对手术中冷冻切片的诊断是不适用的 ,我们应用Ｅｎｈａｎｃｅｄｐｏｌｙｍｅｒｏｎｅ ｓｔｅｐｓｔａｉｎｉｎｇ (Ｅ ＰＯＳ)法进行冷冻切片快速免疫组化染色 ,全部过程可在 13ｍｉｎ内完成 ,对确定或排除肿瘤有明确的辅助诊断作用 ,并能发现周围哨卡淋巴结内的微小转移灶 ,这对淋巴结冷冻切片的诊断尤为重要 ,可指导手术医师确定手术范围。1　材料与方法1.1　材料　手术切除 10ｍｉｎ内的新鲜标本 ,…     

介绍一种改良承载超薄切片制膜法

用于透射电镜的超薄切片往往需要有支持膜的铜网承载,有支持膜就能有效地吸附切片,并能加强切片承受电子束的轰击,但却有影响分辨率、易污染等弊端。近年来,许多电镜工作者采用无膜铜网承载超薄切片,虽能增强切片反差、提高图     

箱式快速石蜡切片法

材料1 木箱1个。规格46×14×16cm,上方为铝板,铝板上取4个直径2.5cm试管孔和1个直径5cm蜡杯孔,前方为活动门。2 容量75ml试管4根。40ml搪瓷     

介绍一种FAAPT冷冻切片快速固定液

手术中冷冻切片的病理诊断是根据冷冻切片组织学形态来进行 ,以确定病变组织的良恶性为目的〔1〕。制作冷冻切片包括制片、固定、染色三个环节 ,在冷冻切片中固定液的选择是不可忽视的一个因素。我们在工作中经反复试验在众多固定液中选择ＦＡＡ液 (福尔马林 -醋酸 -乙醇 ) ,经改良配成一种适合于冷冻切片的ＦＡＡＰＴ(福尔马林 -醋酸 -乙醇 -苦味酸 -ＴｒｉｔｏｎＸ - 10 0 )快速固定液。该液具有固定速度快、渗透力强、ＨＥ染色鲜艳、组织结构清晰等优点。1　材料与方法1 1　ＦＡＡＰＴ固定液的配制　福尔马林 15ｍｌ,醋酸 3ｍｌ,95 %乙醇…     

应用基因芯片检测喉鳞状细胞癌甲醛固定石蜡包埋组织microRNA表达谱

目的 建立以甲醛固定石蜡包埋组织为材料、基于基因芯片技术的microRNA(miRNA)表达谱的分析方法 ;筛选与喉鳞状细胞癌(简称喉癌)生物学特征密切相关的差异表达miRNA.方法 从喉癌甲醛固定石蜡包埋组织中制备总RNA,经质量鉴定后进行荧光标记.采用Agilent公司的容纳723条人类miRNA探针的基因芯片完成杂交实验,以获得喉癌的miRNA表达谱.以GeneSpring GX和R-Project软件处理分析基因芯片实验数据,筛选与喉癌转移相关的差异表达miRNA.结果 从24例甲醛固定石蜡包埋组织标本中获得了符合基因芯片实验质量标准的RNA样品,并完成了基因芯片杂交及数据分析.从中共鉴定到319个miRNA,有96个miRNA在24例喉癌中均有表达,其中与淋巴结转移密切相关的(检错率<0.05)差异表达miRNA有5个,分别为miR-23a* 、miR-28-5p、miR-15a、miR-16和miR-425.结论 甲醛固定石蜡包埋组织可以提供符合基因芯片分析质量要求的miRNA,是研究miRNA的重要样品资源.从喉癌的miRNA表达谱中筛选出的转移相关差异表达miRNA(miR-23a*、miR-28-5p、miR-15a、miR-16和miR-425)有可能成为评估喉癌转移风险的新型分子标志.     

应用基因芯片检测喉鳞状细胞癌甲醛固定石蜡包埋组织microRNA表达谱

目的 建立以甲醛固定石蜡包埋组织为材料、基于基因芯片技术的microRNA(miRNA)表达谱的分析方法 ;筛选与喉鳞状细胞癌(简称喉癌)生物学特征密切相关的差异表达miRNA.方法 从喉癌甲醛固定石蜡包埋组织中制备总RNA,经质量鉴定后进行荧光标记.采用Agilent公司的容纳723条人类miRNA探针的基因芯片完成杂交实验,以获得喉癌的miRNA表达谱.以GeneSpring GX和R-Project软件处理分析基因芯片实验数据,筛选与喉癌转移相关的差异表达miRNA.结果 从24例甲醛固定石蜡包埋组织标本中获得了符合基因芯片实验质量标准的RNA样品,并完成了基因芯片杂交及数据分析.从中共鉴定到319个miRNA,有96个miRNA在24例喉癌中均有表达,其中与淋巴结转移密切相关的(检错率<0.05)差异表达miRNA有5个,分别为miR-23a* 、miR-28-5p、miR-15a、miR-16和miR-425.结论 甲醛固定石蜡包埋组织可以提供符合基因芯片分析质量要求的miRNA,是研究miRNA的重要样品资源.从喉癌的miRNA表达谱中筛选出的转移相关差异表达miRNA(miR-23a*、miR-28-5p、miR-15a、miR-16和miR-425)有可能成为评估喉癌转移风险的新型分子标志.     

应用基因芯片检测喉鳞状细胞癌甲醛固定石蜡包埋组织microRNA表达谱

目的 建立以甲醛固定石蜡包埋组织为材料、基于基因芯片技术的microRNA(miRNA)表达谱的分析方法 ;筛选与喉鳞状细胞癌(简称喉癌)生物学特征密切相关的差异表达miRNA.方法 从喉癌甲醛固定石蜡包埋组织中制备总RNA,经质量鉴定后进行荧光标记.采用Agilent公司的容纳723条人类miRNA探针的基因芯片完成杂交实验,以获得喉癌的miRNA表达谱.以GeneSpring GX和R-Project软件处理分析基因芯片实验数据,筛选与喉癌转移相关的差异表达miRNA.结果 从24例甲醛固定石蜡包埋组织标本中获得了符合基因芯片实验质量标准的RNA样品,并完成了基因芯片杂交及数据分析.从中共鉴定到319个miRNA,有96个miRNA在24例喉癌中均有表达,其中与淋巴结转移密切相关的(检错率<0.05)差异表达miRNA有5个,分别为miR-23a* 、miR-28-5p、miR-15a、miR-16和miR-425.结论 甲醛固定石蜡包埋组织可以提供符合基因芯片分析质量要求的miRNA,是研究miRNA的重要样品资源.从喉癌的miRNA表达谱中筛选出的转移相关差异表达miRNA(miR-23a*、miR-28-5p、miR-15a、miR-16和miR-425)有可能成为评估喉癌转移风险的新型分子标志.     

Investigation of specimen mislabelling in paraffin-embedded tissue using a rapid, allele-specific, PCR-based HLA class II typing method

Mislabelling of surgical specimens can seriously affect the accuracy of histopathology reports. We describe two cases in which clinically suspected mislabelling was investigated by polymerase chain reaction (PCR)-based HLA DRB and DQB tissue typing of paraffin biopsy-derived DNA, using sequence specific primers (PCR-SSP HLA typing). In the first case, two patients underwent surgery for breast carcinoma. A subcutaneous lymph node containing metastatic carcinoma was received with the breast specimen from one patient, but was clinically considered more likely to originate from the other patient who underwent breast surgery on the same day. In the second case, histological examination of retained products of conception from a young woman revealed adenocarcinoma, but a repeat curettage specimen consisted of secretory phase endometrium. In case 1, PCR-SSP HLA typing confirmed the clinical suspicion that the subcutaneous lymph node received with tissue from one patient originated from the other patient. This result converted the first patient from lymph node positive breast carcinoma to lymph node negative disease. In case 2, there was no evidence from PCR-SSP HLA typing that the endometrial samples had originated from different patients. PCR-SSP HLA typing requires fewer steps than methods based on PCR amplification followed by oligonucleotide probing (PCR-SSOP HLA typing), and relies on the amplification of shorter sequences of DNA. Therefore, this technique can produce more rapid results than PCR-SSOP HLA typing, and is ideally suited to typing partially degraded DNA derived from formalin-fixed and paraffin-embedded tissue.     

Multiparameter immunofluorescence on paraffin-embedded tissue sections.

Immunohistochemical techniques have gained increasing importance in diagnostics and research. While formalin-fixed, paraffin-embedded human tissue retains excellent morphology, the detection of antigens by immunofluorescence in its sections and especially the demonstration of multiple simultaneous antibodies have limitations. Double immunofluorescence labeling of routinely processed paraffin sections has been described previously. The signal intensity observed after triple labeling has been reported to be significantly inferior to that obtained by application of double fluorochromes. The authors show multicolor labeling of three and four primary antibodies in routinely processed paraffin-embedded tissue sections using a standardized immunofluorescence technique. In addition, procedures to reduce background staining and to avoid nonspecific double staining are described.     

Immunophenotyping of acute leukemias using paraffin-embedded tissue sections

In a study of 55 patients with either acute lymphoid leukemia (ALL; 25 cases) or acute myeloid leukemia (AML; 30 cases), paraffin-embedded bone marrow particle sections were examined with a panel of monoclonal and polyclonal antibodies reactive toward lymphoid and myeloid-associated antigens, using the alkaline phosphatase-anti-alkaline phosphatase (APAAP) technique. All cases were previously classified according to the French-American-British (FAB) Co-operative Group, and cases of ALL were immunophenotyped by flow cytometry. Results indicated that myeloid-associated antibodies (Mac 387, KP 1 [CD68], antielastase, antilactoferrin, and antilysozyme) did not react with any case of ALL, M1-AML, or M6-AML, whereas at least one of these antibodies reacted with 20 of 21 (95%) cases of M2, M3, M4, and M5-AML. Anti-glycophorin C marked cases of M6-AML, whereas anti-CD3 labeled T-cell ALL. None of the antibodies tested specifically identified cases of B-cell ALL. The authors conclude that use of a selected panel of antibodies on paraffin-embedded bone marrow particle sections may be of value in the diagnosis and immunophenotypic classification of many cases of acute leukemias.     

荧光原位杂交在滑膜肉瘤诊断中的应用

滑膜肉瘤是儿童及青少年期常见的软组织肿瘤,约占软组织恶性肿瘤的2％～10％。好发于大关节周围,也可见于其他关节、软组织,还可发生于肺、前列腺、肾等脏器。组织学可分为双相分化型、单相纤维型、单相上皮型、低分化型H0,其中前2种最常见。由于其形态多样,发病部位广泛,免疫组织化学染色又缺乏特异的抗体,有时会造成病理诊断的困难。细胞和分子遗传学研究发现,90％以上的滑膜肉瘤存在特异的t（X;18）（p11．2;q11．2）,导致位于18号染色体SYT基因易位,和位于X染色体上SSX基因产生SYT—SSX融合基因。目前国内检测该融合基因均使用逆转录-聚合酶链反应（RT—PCR）。我们收集儿童滑膜肉瘤4例,探讨运用荧光原位杂交（FISH）法对甲醛固定、石蜡包埋组织检测其融合基因的可行性。     

Monoclonal antibodies marking B-cell non-Hodgkin's lymphoma in paraffin-embedded tissue

Traditional methods for the immunophenotypic analysis of the non-Hodgkin's lymphomas require fresh or snap-frozen tissue for flow cytometric or immunohistochemical studies. The monoclonal antibodies LN1, LN2, and L26 have been recently developed to recognize B-cell-specific antigens that survive routine tissue processing and paraffin embedding. In this study, the ability of these three antibodies to mark the neoplastic cells in 160 cases of paraffin-embedded non-Hodgkin's lymphoma relative to frozen section immunophenotype (42 T-cell, 118 B-cell), manner of fixation (B5 versus 10% buffered formalin), and histological subtype was examined. With B5-fixed tissue, the percentages of B-cell lymphoma marking with the antibodies were as follows: L26, 96.6%; LN1, 88.2%; LN2, 93.7%. With formalin-fixed tissue, the percentages of B-cell lymphoma reacting with the antibodies were: L26, 89.1%; LN1, 26.2%; LN2, 57.8%. Each of the antibodies marked a small percentage of paraffin-embedded T-cell lymphomas: L26, 4.7%; LN1, 4.7%; LN2, 7.1%. LN2, and to a lesser extent LN1, stained Reed-Sternberg cells, a feature not seen with L26. Nor did L26 mark nonlymphoid neoplasms, a feature previously reported with LN1 and LN2. Since a high percentage of B-cell lymphomas react with these antibodies and they are relatively specific for B-cells, they should prove highly useful for the evaluation of both diagnostic and experimental pathology specimens. L26 offers the distinct advantage of working well in both B5 and formalin-fixed tissues and seemingly not marking epithelial neoplasms.