介绍一款国产冷冻切片机的使用和保养

<正>冷冻切片是临床病理检查中的常用方法,对临床上需要进行快速病理诊断,确定病变性质及手术范围至关重要[1]。随着精准医疗的发展,对术中及术后冷冻切片的质量要求越来越高,冷冻切片的应用也越来越普遍[2]。如何合理的选择、使用及维护冷冻切片机,延长冷冻切片机的使用寿命是病理工作者必需面对的问题。现结合西京医院免疫科于     

恒冷箱切片机对不同新鲜组织冷冻切片的结果分析

恒冷箱切片机是冷冻切片机中最先进 ,最精密、价格昂贵的一种。它用途广泛 ,常用于组织化学 ,特别是酶的组织化学 ,免疫组织化学 ,临床病理切片快速诊断 ,以及某些组织学教学标本的制作[1- 5] 。虽然恒冷箱切片机先进、精密 ,但要做到很好地掌握 ,是一件不易的事。不同的织器官在不同气候环境中它所适宜的切片温度是不一样的。本文对使用恒冷箱切片机作不同新鲜组织进行冷冻切片 ,的最佳条件进行探索 ,为更好地使用恒冷箱切片机提供实施依据。1材料与方法1.1材料恒冷箱切片机 (ＫＲＹＯＳＴＡＴ 172 0型西德ＬＥＩＴＺ公司制造 ) ,动物及…     

全自动冷冻切片机的应用体会

冷冻切片是一种在低温条件下使组织快速冷冻到一定的硬度,然后进行切片的一种技术.随着临床的发展,手术中需要快速病理诊断的病例越来越多,冷冻切片技术对确定手术方案具有积极的临床意义[1].冷冻切片质量的优劣直接关系到病理诊断的准确率,做1张好的切片,除了技术过硬的技术员外,冷冻切片机也起着重要的作用[2,3].本文通过摸索实践,能够使各类组织的快速制片质量达到常规.现将南通市肿瘤医院病理科使用的德国Leica CM1950冷冻切片机应用体会介绍如下.     

冷冻切片机的使用及消毒保养

快速冷冻切片法是一种在低温下借助OCT水包埋组织,短时间内达到一定硬度,在恒冷冷冻切片机内进行切片的方法。冷冻切片机已经得到广泛使用,为加强仪器管理,现就冷冻切片机的使用及消毒保养要点进行探讨,以便更好地为临床诊疗服务。     

提高乳腺冷冻切片制作质量的体会

术中制作快速冷冻切片是目前术中诊断乳腺疾病的主要病理学检查方法之一.因乳腺标本中脂质含量高,制片难度较大.本文现将提高乳腺冷冻切片制作质量的体会介绍如下. 1减少冰晶的形成 冰晶的形成与结晶核的数目和晶体形成的速度有一定关系.利用较低的温度使组织迅速冷冻,结晶核形成快而多,使冰晶体积变小[1].乳腺组织中脂质和水分含量较多,为避免和减少冰晶形成(图1)我们建议:(1)采用急骤冷冻的方法使组织在低温下迅速冷冻.可将组织放在冻托上置入液氮中,10～20 s后取出,置于低温恒冷箱切片机中切片.     

恒冷切片箱对固定后组织的冷冻切片技术

恒冷切片箱是冷冻切片机中最先进 ,最精密的一种。它用途广泛 ,日前常用于组织化学特别是酶组织化学、免疫组织化学、临床病理切片快速诊断、以及一些组织学教学标本的制作。以往一般都是用新鲜组织直接行冷冻切片 ,虽简易方便 ,速度快 ,但新鲜组织经冷冻 ,组织细胞内常形成冰晶 ,造成一定程度的损伤 ,影响结构的完整性 (图 1) ,尤其在免疫组织化学研究中 ,很容易使组织结构不清 ,抗原定位不精确 ,影响了观察和结果[1、2 ] ,笔者经过多年摸索和实践 ,采用组织先固定后行冷冻切片 ,并将固定组织经鸡蛋白甘油浸渍处理 ,克服了组织细胞冰晶的形…     

病理冷冻切片制作质量的探讨

目的 探讨病理冷冻切片制作的质量.方法 严格规范操作取材、切片、固定、染色、封片等制片过程中的各个环节,对制片质量进行观察.结果 经光镜观察切片,切片组织切面完整,厚薄均匀,效果良好,所制切片色彩鲜艳,对比清晰.结论 只有严格规范地做好制片中各个环节的技术操作,才能避免冷冻切片制作中出现各种常见问题,提高冷冻切片质量,为临床医师决定手术方式、范围,缩短手术时间,确定进一步治疗方案,提供可靠的诊断依据,对临床治疗具有重要意义.     

乳腺手术中细胞印片的病理诊断价值

手术中快速病理诊断方法包括冷冻切片、快速石蜡切片和细胞学诊断。由于冷冻切片机设备的不断改进,使冷冻切片质量有了保证,加上具有操作时间短等优点而被广泛应用。快速石蜡切片由于操作步骤较烦琐,时间较长而很少被应用。细胞印片技术操作简单,在更短时间内即可对所送标本做出定性诊断。我们通过对772例术中乳腺标本分别进行细胞印片和冷冻切片检查,并对二者进行比较,探讨细胞印片法在乳腺块中的应用价值。     

用已固定组织制作冷冻切片体会

经福尔马林固定的组织不能直接做冷冻切片 ,为此作者经过反复试验 ,探索出一种利用液氮快速冷冻制作冷冻切片的方法 ,取得了满意的效果 ,现介绍如下。1　材料和方法　　经常规福尔马林固定 1～ 2天的外检和动物标本 (骨和含钙组织除外 ) ;低温恒冷切片机、ＯＣＴ包埋剂、包埋托、涂有防脱片胶的载玻片、液氮一罐。将福尔马林固定的组织常规取材 ,用滤纸吸干周围水分 ,置包埋托上加ＯＣＴ包埋剂 ,入液氮 3～ 5ｍｉｎ后取出 ,置于冷冻切片机冷冻台上片刻 ,修切组织 ,切片 ,吹干 ,不需固定 ,直接ＨＥ染色。2　体会　　组织从液氮中取出不宜立即…     

酵素水溶液在冷冻切片染色分化中的应用

冷冻切片是保证术中病理诊断和进行科学研究的重要手段,切片质量的好坏直接影响快速诊断的结果及科研的可靠性。影响冷冻切片质量的因素很多,分化液是其中之一。酵素又称为酶,具有催化、净化等作用,现今在生产、生活中广泛应用,如清洗、灭菌、净化、美容、减肥等。有关其在病理切片制作中的应用尚未尝试。作者在不同组织冷冻切片染色中利用酵素水溶液分化,以期探索出提高切片染色清晰度、对比度,优化冷冻切片质量的方法。     

Measurement of section thickness in quantitative microscopy with special reference to enzyme histochemistry

Quantitative histochemical techniques demand constancy in sectioning tissue and preferably knowledge of the true thickness of a section. In this investigation a simple, rapid, accurate, and reproducible technique is described for measuring section thickness using an instrument known as a Surfometer. It has been shown that: (a) cryostat microtome settings bear little relationship to the true section thickness of the sections produced; (b) that different instruments differ in their inaccuracies; (c) that this inaccuracy varies slightly with different tissues.If densitometric methods are used and either the absolute amount of a tissue component is to be measured or the amount of a tissue component is to be compared in two tissues then it is mandatory to determine the actual thickness of the section. The method described here seems the most appropriate one for these investigations.     

Cryosectioning of undecalcified tissues for immunofluorescence.

The present report describes a procedure for preparing 4--6 micrometers cryostat sections of undecalcified fresh frozen tissue which contain hard tissue, for immunofluorescence. The apparatus used is a cryomicrotome originally designed for cutting sections for whole body autoradiography. To obtain cryostat sections suitable for tissue immunofluorescence the standard procedure was modified with respect to the hardness and edges of the microtome knife, the temperature of the cryostat and the carboxymethyl cellulose concentration of the embedding material.     

Use of microwave oven improves morphology and staining of cryostat sections.

The quality of microscopic image of cryostat sections that had been subjected to microwave assisted fixation was compared with that resulting from conventional air drying of the sections. The role of microwaves in producing rapid special stains on cryostat sections was also assessed. Methods used permitted stains such as periodic acid Schiff, alcian blue, Gordon and Sweets's reticulin, Masson Fontana, Elastica, Prussian blue and Van Gieson to be performed within three minutes of cutting a cryostat section. The cytological detail of nuclei was much clearer using the microwave technique, allowing more accurate determination of cell type. The microwave oven seems to have major potential in improving the diagnostic accuracy of surgical frozen sections.     

Acetone/periodate-lysine-paraformaldehyde (PLP) fixation and improved morphology of cryostat sections for immunohistochemistry

Cryostat sections are extensively used in the study of lymphoid tissues because the majority of antigens do not survive routine fixation and processing. A major disadvantage of cryostat section immunohistochemistry is their poor morphology. The use of periodate-lysine-paraformaldehyde (PLP) fixation regimes for cryostat sections was evaluated and compared with conventional acetone immersion. This fixation gives excellent morphology but poor immunostaining. The quality of immunostaining with a panel of 31 antibodies was good with brief acetone fixation followed by PLP fixation. Morphological preservation was very good. We suggest that acetone-PLP fixation is an improvement over acetone alone for cryostat section immunohistochemistry.     

A possibility of cutting frozen sections of bone tissue in routine pathology

A low priced, efficient frozen section cutting device is described to produce 5 microns sections of frozen undecalcified bone specimen blocks in routine pathology. The device was developed by combining two commercially-available instruments, namely a frozen section microtome with methyl alcohol cooling and a hard tissue microtome. Bore holes were drilled in the hard tissue knife to permit cooling, using the methyl alcohol system of the other microtome.     

A method of preparing sections for express histological diagnosis

The method of improving quality of histological slides for intrasurgical diagnosis is proposed. The method consists in preliminary microwave processing of the slice of surgical material before putting in into cryostat for freezing and sections preparation. A slice of surgical material was first immersed in 10 ml of physiological solution (pH = 7.2). Then it was exposed to microwave radiation (2.45 GHz, 1 W/cm2) for 1.5 min. After that the slice was put into cryostat and frozen. Microwave processing is done in a home microwave oven or working chamber of microwave histoprocessor. Essential advantage in quality of slides is obtained.     

Use of the cryostat section in electron microscopy.

A method using the cryostat section as a source of material from which relevant areas for ultrastructural examination can be positively located and quickly processed is described. Preservation of the tissue does not appear to be unduly affected by freezing and subsequent cryostat sectioning.     

Histological and cytological criteria in the diagnosis of malignant melanomas by cryostat sections

Summary Although cryostat sections in general allow a distinction to be made between malignant melanomas and other pigmented lesions in clinically doubtful cases, the differential diagnosis may be difficult. The histological and cytological criteria taken into account can be classified as major, minor, and insufficient. Knowing the diagnostic value of each makes a conventionally established diagnosis safer. Variance analysis does not contribute to the problem but it can nevertheless be shown that the evaluation of six major criteria makes a quick and reliable cryostat section diagnosis possible. If these results are confirmed in a prospective study it would be a decisive step on the way to a quicker and safer cryostat section diagnosis of malignant melanoma, even for the less experienced histopathologist.The results published here were presented in part at the DDG meeting 1980 at Westerland/SyltWe are grateful to Miss Schubert, Institute of Biomathematics of the University of Munich, for the statistical evaluations     

Application of domestic microwave for urgent histopathology reporting: an evaluation

Rapid diagnosis of histopathological material is becoming increasingly desirable. In neuropathology, crush smear preparation and frozen section diagnosis of tissues removed during operative procedures, have remained as essential tools for rapid diagnosis. Microwave technology has been introduced into the field of tissue processing and staining in past decade. Now-a-days even automated microwave assisted rapid tissue processors are available. In our study we have analysed the use of a domestic microwave (cost approximately Rs.5000) for urgent histoprocessing (30 minutes). This could be useful in small laboratories or the ones which are in the phase of establishing the department as the procedure is much more economical than obtaining a frozen section (which requires a cryostat worth 3-6 lakhs) and the interpretation of the section obtained does not require any extra experience as these resemble the routinely processed tissue sections. The advantages and limitations of the procedure have been discussed.     

Electron microscopical enzyme histochemistry on unfixed tissues and cells. Bridging the gap between LM and EM enzyme histochemistry

In principle, enzyme histochemistry should be performed on unfixed tissues and cells to avoid inhibition of enzyme activity by chemical fixation. For EM enzyme histochemistry, unfixed tissue specimens include fresh tissue blocks, non-frozen tissue chopper sections, cryostat sections and cell preparations. Studies on localization of enzyme activity at the ultrastructural level in unfixed specimens, be it fresh or frozen, are reviewed here. Preservation of ultrastructural morphology is discussed with special attention to the effects of freezing. It is concluded that unfixed cryostat sections are the best alternative for EM histochemistry of tissues, when interposing a semipermeable membrane in between cryostat section and gelled incubation medium. It is an adequate method to preserve structural integrity of unfixed tissue on the one hand and to avoid inactivation of the enzyme by chemical fixation on the other. For EM cytochemistry on individual cells, a better preservation of ultrastructure may be obtained because freezing can be avoided, but mild pretreatment with a fixative or detergent may be necessary to permeabilize cellular membranes for demonstration of intracellular enzyme activity.