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merck millipore 默克密理博纯水超纯水一体系统明澈D24UV操作手册

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明澈-D 24 UV
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物品检查清单安装开始前请核实所有物品是否可用。
o 纯水系统主机 (A) o 用户手册和快速参考指南 (B) o 外部电源 (C) o PROGARD®预处理柱和 Q-GARD®超纯水柱（D） o 储水水箱：30、60 或 100 升（E） o POU 取水器（F） o 包含下述的附件包：管材配件电源
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2 显示器和小键盘
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3 Progard® 预处理柱位置
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4 锁定手柄
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5 消毒/清洁口
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6 电源接口
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7 系统进出水口连接
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*系统断电时，应将电源与插座断开。
说明系统 ID 和序列号电源适配器* Q-GARD®超纯水柱位置超纯水进出口接口 POU（用水点）取水POU 取水开关取水器终端过滤器
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您的纯水系统不适于家庭使用。
纯水系统长时间断电会使计时器的电池放电，导致失去时间和日期记录。如果长时间关闭纯水系统，应使用实验室关闭模式请与您的服务代表联系。
经常保存系统的历史记录-每三个月或每半年。参见 Millitrack 文档。
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Merck Millipore 30L-100L水存储胶囊说明书

Storage Tanks and AccessoriesStorage with a DifferenceMerck Millipore is a business ofGuarantee the purity of your stored waterPure water requires a storage system to prevent the degradation of your water quality. Merck Millipore’s 30-, 60-, and 100-liter polyethylene (PE) storage tanks are designed to maintain consistent purity of stored water and provide effective protection against airborne contaminants.*Prevent contamination Water stagnancy can cause bacterial proliferation. Our optimal Automatic Sanitization Module (ASM) provides the ideal solution for the prevention of bacterial growth and biofilm formation on the inner surface of the storage tank. In addition, our advanced vent filter protects pure water from airborne contamination.Distribute your stored water where it is neededTo provide pure water for use with all of their applications, laboratories need to be able to distribute stored water from their water purification system storage tanks.- For distribution of non-pressurized pure water,conveniently located on the front of the Merck Milliporestorage tanks.- For convenient distribution the storage tank, an E-POD® point-of-delivery dispenser can be connected to an Elix® Advantage or Milli-Q® Integral water purification system.- For automatic feed of pure water, distribution valves on the base of the storage tank allow connection to other laboratory equipment such as glassware washing machines. Distributionpumps are also available if needed.* A complete line of storage tanks is available, ranging in capacity from a few liters to several hundred liters . Your nearest Merck Millipore office will be able to guide you in the choice of the tank best suited to your needs.4562Unique features• Polyethylene selected for its minimum release of extractables • Opaque walls block sunlight to prevent algae development • Smooth inner surface prevents biofilm formation• Cylindrical shape minimizes surface area in contact with water • Conical bottom allows complete draining for cleaning and rinsing • Pure water smoothly fed in at the bottom of the tank prevents absorption of carbon dioxide• Front valve enables manual dispense of pure water• Distribution valves permit connection to other laboratory equipment • Hermetically sealed lid blocks air from entering the tank• Large top opening allows manual cleaning during sanitization procedure • Compact space-saving designFail-safe protection• Sensor rod float switch system for automatic storage tank refill and indication of water level (% full)• Overflow connected to the drain, in the unlikely event of a water system malfunction• Direct display of stored water level on water purification system units • Water sensorMerck Millipore 30-, 60-, and 100-liter polyethylene storage tanks incorporate the latest technical developments and advanced features for stored water of consistent purity.All tanks have a small footprint and are designed for wall-mounting if required. Underbench installation is also possible for some models.For details of the tests performed during the storage tank development process, please request the publication “R &D Notebook 1: Optimizing the storage of purified water for laboratory applications” (Ref. No.: RD001EN00)from your local Merck Millipore representative.Optimized pure water storageInnovative storage tank designStorage tanks designed for efficiencyThe main concern when storing pure water is degradation of water purity over time. Only a strict choice of storage tank materials, associated with a careful design and appropriate protection against airborne contaminants, can ensure consistent water quality during storage.Designed for efficiency1. Hermetically sealed lid2. Sensor rod float switch3. Sanitary overflow4. Blow-molded storage tank5. Cylindrical shape6. Conical bottom with distribution valves7. Front dispensing valve8.Advanced vent filter3Advanced storage tank vent filterStorage Tank AccessoriesIn order to help ensure optimum purity and distribution of your stored water, Merck Millipore offers a range of accessories and connections for your storage tank, including the following items:Advanced vent filter:important protection against airborne contaminantsAir is contaminated by carbon dioxide, particles, microorganisms, and volatile organic compounds that come mainly from the laboratory atmosphere. To protect pure water from all these contaminants, Merck Millipore has developed an advanced storage tank vent filter that includes:• Activated carbon to adsorb volatile organics (including lab solvents such as acetone, chloroform, and methanol)• A soda-lime bed to remove CO 2• A Durapore® hydrophobic membrane for particle and bacteria retentionThis advanced vent filter is recommended for the protection of high-resistivity water, such as Elix® product water, during storage. To protect RiOs™ reverse osmosis-quality water, a Durapore® 0.45 µm hydrophobic membrane vent filter is also available.1. Airborne Contaminants A. Volatile Organics B. Particles C. Bacteria D. CO 22. Volatile Organics absorption3. CO 2 removal4. Particle and Bacterial retention5. Storage tank inletPurified air enters the storage tank• Advanced Vent Filter• Automatic Sanitization Module • Air Gap Device• E-POD® pure water remote dispenser • Washer Distribution Kit • Water Sensor • Lab Close KitComplete Merck Millipore water purification chain with the ASM and water sensor4Automatic Sanitization Module (ASM):say “No!” to bacterial proliferation Maintaining high purity water with low bacteria levels during storage is critical. If left to proliferate, trace levels of microorganisms present in pure water compromise water purity. This bacterial contamination is responsible for the formation of a biofilm — an accumulation of organic material made up of active and dead organisms, on the inner walls of the storage tank.Even though chemical sanitization and mechanical scrubbing may be periodically performed, this biofilm is difficult to remove and is a frequent source of recontamination in stored water.Merck Millipore’s ASM is designed to prevent the growth and proliferation of bacteria and the resulting biofilm on the inner surface of Merck Millipore PE storage tanks. The ASM makes use of the germicidal properties of an ultraviolet (UV) light at 254 nm, which is fitted inside the tank.The ASM provides full flexibility for guaranteed results• 254 nm UV lamp; selected for its germicidal effectiveness • Pre-programmed intervals of 10-min / day automatic UV illumination for optimized efficiency• Additional programmable and manual UV exposure possible to meet critical application requirements• Up to 45 min /day of UV exposure for total flexibility• Program daily time settings, UV cycles, and UV lamp operation displayed on the Millitrack® e-Solution dashboard • UV lamp exchange alarm for easy maintenance• Compact design allowing installation on top of the storage tank10 minutes of daily UV exposure is sufficientDuring development of the ASM, the UV lamp exposure cycles were optimized by examining the resulting bacterial reduction after exposure.Two 60-liter storage tanks were fed by an intentionally contaminated reverse osmosis water purification system. The tanks were then emptied and refilled each day and alternately equipped with an ASM into which variable illumination times were programmed.As shown in the graphs, 10 minutes per day of UV exposure were enough to make the tanks return to their original low bacterial levels.Biofilm formation over timeGermicidal effectiveness of the UV lamp5Air gap device for protection against bacteriological contaminationWater purification systems and storage tanks sometimes require a connection to the drain. Drains are typically dirty environments contaminated by microorganisms, and in particular, bacteria. Therefore, when the outlet of the water system reject tubing is pushed into the drain, there is a risk that bacteria could contaminate the inside of the reject tubing, and then progressively move to the water system.One way to prevent this from occurring is to install an air gap device on the reject tubing. This allows the reject water flow to movethrough the tubing without touching the inside of the contaminateddrain environment. Installing an air gap device is an easy and safe way to prevent the development of bacteria above the air gap level.The E-POD® Elix® water point-of-delivery unit can be connected to an Elix® Advantage pure water system or Milli-Q® Integral pure and ultrapure water system to dispense pure water wherever it is needed in the lab.Advantages of the E-POD® remote dispenser include: • Improved bacterial water quality(less than 0.1 cfu/mL, with final filter)• V ersatility enabling use for multiple applications or users when a Millipak® or Biopak® polisher is fitted to the dispenser outlet• Volumetric dispensing to save time • Ergonomic design and ease of use• Flexibility, with installation of up to three E-POD® units per system• Information at a glance thanks to the color backlit screen on the dispenser base •Space-saving small footprintE-POD® pure water remote dispenserAir Gap DeviceE-POD® pure water remote dispenser: pure water where you need it6Water sensor for control over feed water supplyIf there is water on the floor, the water sensor enables shutdown of the feed water supply in order to prevent a lab flood.Lab Close Kit keeps your system in top condition when you’re away When your facility is closed for an extended time — such as vacation periods — the Lab Close K it will avoid water purification system standstill during these long periods. The Lab Close K it intelligently optimizes the consumption of water and electricity by your system, preventing the negative effects of nonuse, such as bacteria buildup. Your water purification system remains in top condition, ready forimmediate use upon your return.Distribution pumps to meet increased demandsWasher Distribution KitA key use of pure water is as feed to laboratory appliances such as glassware washers, autoclaves, sterilizers and weathering devices.The Merck Millipore Washer Distribution Kit provides cost-effective and convenient distribution of pure water to common laboratory appliances, with flow rates between 12.5 – 13.5 L/min (at 2 bar or 30 psi, depending on voltage).Installation of the small-footprint kit is fast, easy, and universal, with bench, underbench, or wall-mounted options. Users profit from a silent, automatic supply of pure water when required.The graph shows characteristics of Merck Millipore distribution pumps.Pumps provide a long service lifetime and quiet operation.Washer Distribution KitSpecifications & Ordering InformationSpecifications for Polyethylene Storage TanksThere are several different storage tank sizes available in this range:* For 30-liter and 100-liter storage tanks, underbench models are also available.Ordering InformationPolyethylene Storage Tanks7For more information, please visit our website:/labwaterPB5506EN00 Copyright 2015 EMD Millipore Corporation, Billerica, MA., U.S.A.Merck Millipore, the M mark, Milli-Q, Elix, AFS, E-POD, Millitrack, Millipak, Biopak, and Durapore are registered trademarks of, and RiOs is a trademark of, Merck KGaA, Darmstadt, Germany.All rights reserved.。

流式细胞术最全攻略：从protocol到问题解决

流式细胞术最全攻略：从protocol到问题解决文献中我们经常能看见这种图，也就是流式结果分析图，美观、简单、一目了然。

流式细胞术（flow cytometry）是20世纪60年代后期开始发展起来的利用流式细胞仪快速定量分析细胞群的物理化学特征以及根据这些物理化学特征精确分选细胞的新技术，主要分为流式分析和分选2部分。

今天我们主要介绍流式分析中基本操作与技巧，首先简要了解一下什么是流式细胞术。

一、流式细胞术的3大要素1. 流式细胞仪：现在市面上有多种型号的流式细胞仪，但其基本结构都是相同的，分析性流式细胞仪有液流系统、光路系统、监测分析系统。

（1）液流系统（2）光路系统光路系统始于激光器，其分类分法很多，最常用的分类方法是根据其发射的激光的波长来分，如常见的有488nm的蓝激光器、635nm的红激光器和405nm的紫激光器，不同的激光器发出的激光照射到细胞后产生的光信号会经过不同的光路系统被不同的通道接收。

流式细胞仪采集的光信号包括散射光信号和荧光信号，散射光信号也就是我们常说的FSC（前向散射光，反应细胞的大小）、SSC（侧向散射光，反应细胞的复杂程度）；荧光信号是细胞上结合有荧光素，被激光激发以后，会发射荧光信号。

（3）检测分析系统：流式细胞仪的检测分析系统就是以通道为单位将细胞的各个通道的光信号汇总分析，最后得出样品群体中细胞的物理化学特征。

通道，我们可以理解为就是光电倍增管，有2个作用，①将光信号转变为电子信号；②放大电子信号。

可以分为散射光通道（FSC通道、SSC通道）和荧光通道，一个激光器下可以有多个荧光通道，一个通道对应多个荧光染料，例如以beckman的DXflex机器的638nm 红激光器为例，APC通道可以接收APC、Alexa Fluor647、eFluor660荧光信号，原则上，这3种荧光素不能同时标记一个样品，否则，就无法分析该通道上的信号是来自于哪种荧光素。

另一个重要组成部分就是计算机分析系统，例如BD的DIVA系统。

细胞活性的检测方法比较

细胞活性的检测方法比较简介：细胞活性是进行细胞生物学实验的一项重要指标，是评价细胞培养、细胞毒性及生理研究的基础。

一些基于细胞的下游分析实验，需要前期有好的细胞，才能保证下游的实验结果的可信度以及实验的可重复性，因此，细胞活性的实验具有广泛的应用。

目前关于细胞活性的检测方法多种多样，如基于台盼蓝染色的细胞计数，以及在此基础上的图像分析型的计数设备，或者是基于荧光染料标记的流式细胞术的分析等。

本文将一种新的检测方法与传统的检测方法进行比较，考察不同方法之间数据的差异性与方法学本身的操作性和成本进行比较。

细胞活性的定义具有多样性，通过对关键的细胞生理指标进行标记来进行检测，如细胞代谢的活性（酯酶的功能，MTT法）、凋亡标记物（Annexin V）、细胞氧化还原电位、膜电位、增值率（DNA含量）、线粒体功能和膜的完整性等。

虽然有如此多的指标参数，但是基于对细胞膜的完整性和相关的染料排斥发检测，已经被广泛的认为是作为细胞活性检测的标准。

应用最广泛的细胞膜完整性的检测方法是台盼蓝染色法，该法最早是基于显微镜下观测并且手工计数，过程费事费力，并且计数的系统误差较大。

为了排除人工计数的误差，增加计数过程的准确性，出现了基于仪器放大、拍照计数的方法。

这样提高了计数过程的自动化程度，但是依旧无法摆脱由于计数时焦平面的选择、颗粒识别的准确性问题，并且容易高估细胞活性。

为了解决这些问题，一些荧光染料被发明进行使用。

该法能够较台盼蓝染色提高100倍或者更高的检测准确性与灵敏度。

这类型的荧光染料中，碘化丙啶（Propidium Iodidi，PI）是应用最广泛的一种标准染料。

与其他基于荧光染料标记的实验一样，该法的缺点就是需要使用荧光显微镜或者是流式细胞仪。

前者要求配置一套荧光显微镜，相对应的激发光源以及滤光片，并且操作过程依旧是手工操作，人工计算实验结果，过程耗时还无法避免人为的主观因素。

而流式细胞仪能够准确的计数，并且进行统计学分析，但是设备昂贵，需要有经验的实验人员进行专门操作，以及需要一定的设备维护等问题。

流式细胞分析技术

ü 光电倍增管（PMT）：
检测散射光和荧光；同时将光学信号转换成电脉冲信号。
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滤光片
分色反光镜
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PMT 滤光片
荧光和散射光检测系统
分色反光镜
激发光
激光聚光系统
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数据处理系统
• 计算机及其软件组成 • 进行实验数据的分析、存储、显示
Photons/Detector (V)
光信号转化为电脉冲信号
特点：单细胞、快速、高通量、多参数、准确、灵敏
经典流式细胞仪（分析型和分选型）量化成像分析流式细胞仪质谱流式细胞仪
单激发光；4个荧光检测通道
经典流式细胞仪
双激发光；6个荧光检测通道
Coulter EPICS XL
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量化成像分析流式细胞仪
美国merck millipore公司
质谱流式细胞仪
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经典流式细胞仪
液流系统-液流聚焦原理
喷嘴
鞘液(Sheath）
喷嘴
荧光信号或侧向散射光
样本流
前向散射光单个细胞流
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液流系统形成单个细胞流示意图
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光路系统
ü 激发光：
常用的是空冷式的氩离子激光光源（488nm）；氦氖激光光源（633nm)；紫外光源
ü 各种光学镜片：
分色反光镜光束成形器；透镜滤光片：长通滤片；短通滤片；带通滤片
(一）标本采集、运输、保存和操作 l标本来源
l几乎所有组织细胞均可用于FCM检测 l免疫标本主要来源于外周血、骨髓、淋巴器官或组织等
l抗凝剂选择
l肝素钠(首选) lEDTA-Na2
l标本保存
标本制备
ü 外周血或骨髓样本

流式细胞仪常用技术方法)

流式细胞仪常用技术方法细胞周期/DNA分析(PI法)一、鞘液准备(至少3L PBS,提前一天准备)0.01M的PBS配制方法:800ml蒸馏水中溶解NaCl8.0g;KCl0.2g;Na2HPO40.24g;调节pH到7.2-7.4(用HCl或NaOH调节,各地蒸馏水的酸碱度不同),加蒸馏水定容至1L,0.22um滤膜过滤后,室温保存。

二、制备一个阴性对照来设定流式细胞仪的取样参数和十字门的范围:没有染色的细胞(制备过程如下,仅不加抗体)三、实验步骤1.取对数期生长细胞,倒去培养液,胰酶适度消化细胞,用培养液吹打,1000rpm,离心10min弃上清;2.PBS洗2次,加0.5ml吹匀,务必吹散;3.加入70%预冷乙醇中(标准做法是将细胞悬液加入预冷70%酒精),固定时可加入Triton X-100以增加膜的通透性)封口膜封口,4℃固定1-2h或过夜(可长至一个月);4.1000rpm×10min离心收集细胞,PBS洗两次;5.用0.4mlPBS重悬细胞并转移至离心管中轻轻吹打;6.加入Rnase-A约3ul至终浓度50ug/ml,37℃水浴消化30min;7.加入PI约50ul至终浓度约为5-50ug/ml,室温避光染色30min;8.用300目滤网过滤,上机检测。

细胞周期/DNA分析(BD公司CycleTEST PLUS DNA Reagent Kit)一、鞘液准备(至少3L PBS,提前一天准备)0.01M的PBS配制方法:800ml蒸馏水中溶解NaCl8.0g;KCl0.2g;Na2HPO40.24g;调节pH到7.2-7.4(用HCl或NaOH调节,各地蒸馏水的酸碱度不同),加蒸馏水定容至1L,0.22um滤膜过滤后,室温保存。

二、制备一个阴性对照来设定流式细胞仪的取样参数和十字门的范围:没有染色的细胞(制备过程如下1-6)三、若做DNA倍体分析,需另设附加对照管肿瘤细胞和外周血单核细胞的混合管,比例至少为2:1四、实验步骤1. 将细胞消化后,用冷PBS洗细胞两次,300g×5min(若细胞数过少可提高转速到500g),为减少细胞损失可用1.5ml离心管离心;2. 弃上清留大约50ul下面的液体防止碰到细胞团,加入1ml Buffer Solution并低速混匀细胞。

流式细胞仪使用一般方法及技巧

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TITER
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10
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Dilution
孵育、溶血、固定
• 抗体孵育：室温下避光15分钟（某些情况下如：细胞
内因子检测需冰育） • 溶血：如样本含中有红细胞需溶血（见下页） • 样本溶血后需立即上机检测，固定后可放置较长时间
白细胞保护型溶血剂Cal-Lyse(其中已含细胞固定剂)：
标本处理时间：
新鲜样本分析时，样本采集后应立即分析
样本固定方法：
1%的多聚甲醛或75%的酒精，作用30分钟。注：不同的实验，所用的固定方法不完全相同。
样本处理标准试剂：
一、10%Bovine Serum Albumin BSA
1. 溶解10g BSA至100ml蒸馏水中 2. 4º C，20000 g离心30分钟
APC
• 激发光633nm，发射光660 在配有双激光的流式细胞仪上，此荧光染料与FITC、PE、 PE-Cy5联用做四色分析。但现在因单激光四色可实现，故使用该染料意义不大。
核酸染料
染料 Propidium Iodide Ethidium biromide 7-Aminoactinomycin D Acridine Orange Chromomycin A3 Hoechst 33342 DAPI Pyronin Y Thiazole Orange YO-PRO-1 TO-PRO-3 LDS 751 激发光发射光 495342 639 493320 637 546 647 503 530(DNA) 640(RNA) 430 580 395 450 372 456 545 565 509 533 642 661 642 661 543 712 光源 488 488 514488 488 457 UV UV 514488 488 488 630 488

Millisolve 清澈系统用户指南说明书

User GuideMillisolve™ Clarification System Cat. No. XX1614700IntroductionThe Millisolve™ Clarification System is designed for vacuum filtration of liquid chromatography (LC) buffers and solvents. The system uses a disposable 0.2/0.22 or 0.45 micron (µm) filter to remove particles which can shorten column life. Vacuum filtration through this system can also remove a large portion of dissolved gases from buffers, reducing the risk of air bubble interference with LC instrumentation.NOTE: To avoid solvent and buffer cross-contamination, EMD Millipore Corporation recommends a dedicated filtration system for each solvent and use of a new filter to process each batch of solvent.The Millisolve™ Clarification System is supplied with two different filter holder configurations. For hazardous fluids, the vacuum cap configuration allows filtration to take place in a closed system. When vacuum is applied, fluid is drawn directly from a bottle, through the filter, and into the conical Millisolve™ flask. For batch filtration, fluid is poured into the open 300 milliliter (mL) funnel and flows through the filter and into the Millisolve™ flask when vacuum is applied. The following table shows the types of membrane filters appropriate for filtering various solvents and buffers.Type of Solvent or Buffer Filter to Use Pore Size Cat. No.Water, buffers MF-Millipore™ hydrophilic mixed celluloseesters filter (Triton® surfactant-free)0.22 µm0.45 µmGSTF04700HATF04700Nylon hydrophilic membrane filter0.2 µm0.45 µm GNWP04700 HNWP04700Omnipore™ hydrophilic polytetrafluoroethylene (PTFE) filter0.2 µm0.45 µmJGWP04700JHWP04700LCR hydrophilic PTFE filter0.45 µm FHLC04700Water, buffers, alcohol, or mixtures Durapore® polyvinylidene fluoride (PVDF)hydrophilic filter0.22 µm0.45 µmGVWP04700HVLP04700 Nylon hydrophilic membrane filter0.2 µm0.45 µmGNWP04700HNWP04700 Omnipore™ hydrophilic PTFE filter0.2 µm0.45 µmJGWP04700JHWP04700 LCR hydrophilic PTFE filter0.45 µm FHLC04700All organic solvents (methylene chloride, hexane, tetrahydrofuran)Fluoropore™ hydrophobic PTFE filter0.2 µm0.45 µmFGLP04700FHLP04700orFHUP04700 Omnipore™ hydrophilic PTFE filter0.2 µm0.45 µmJGWP04700JHWP04700 LCR hydrophilic PTFE filter0.45 µm FHLC04700Additional Equipment Required●47 millimeter (mm) filter appropriate for your application●Vacuum source, either central laboratory vacuum or a portable vacuum pump. See Ordering Information foravailable models.Y WARNING: Choose a vacuum source appropriate for the fluid being filtered. Many vacuum pumps are NOT explosion-proof and should not be used when filtering flammable or explosive liquids.●Vacuum tubing, 3/16 inch (in.) (4.8 mm) inner diameter (ID) silicone rubber (cat. no. XX7100004)Millisolve™ Clarification System ComponentsVacuumNOTE: Smooth-tip filter forceps are supplied but not shown.A vacuum base with a stainless steel screen filter support (instead of a coarse-frit glass support) is also available(cat. no. XX1514732). The stainless steel screen filter support is easier to clean, and improves the filtration rate. PrecautionsY WARNING: Filtration of flammable liquids can generate enough electrostatic charge to cause a fire or explosion. To mitigate this potential, follow these precautions:●DO NOT use the stainless steel screen filter support with highly flammable solvents.●Work under a hood (or another ventilation system) when filtering flammable liquids toprevent accumulation of ignitable vapor and air mixtures. Ventilation systems, includingfume hoods, must be designed in accordance with the National Fire Protection Association(NFPA) code.●Keep the filtration rate low; apply only enough vacuum pressure to maintain steady flowthrough the filter.●Follow safe handling guidelines for laboratory glassware when working with the Millisolve™ Clarification System.Information on safe handling of glassware can be found at /labware/glassware.●Although this product is rated for 948 mbar (28 in. Hg), it is not necessary to apply maximum vacuum pressure toachieve a reasonable filtration rate. Whenever possible, operate at lower vacuum pressure in order minimize the potential for glassware breakage due to implosion.●Before use, soak new glassware in 1% hydrochloric or nitric acid for several hours, then wash and rinse thoroughly with laboratory grade water . This neutralizes the slight alkalinity of the new glass and ensures that loose particles (glass beads) are removed from the fritted glass filter support. Refer to the Cleaning section for details on cleaning after use. ●To protect the vacuum source from fluid contamination, install a vacuum trap flask between the Millisolve™ flask and the vacuum source and place a Millex ®-FA 50 filter (cat. no. SLFA05010) in the vacuum line between the vacuum trap flask and vacuum source.Millisolve™ system Vacuum trap Vacuum pump●When using the vacuum cap for continuous filtration, make sure that the filter is dry. If the filter is wet at the beginning of the filtration process (before the solvent/buffer flow enters the vacuum cap), there will be no flow because the vacuum pressure is not high enough to overcome the filter bubble point. If the filtration process is interrupted or air is introduced into the line, use a new, dry filter on a dry support to restart filtration.Operation1. Connect one end of the vacuum tubing to the sidearm (outlet) of the vacuum base. Push gently to secure thetubing, wetting the sidearm with water , if necessary.2. Optional : Stretch the funnel alignment guide over the bottom ofthe vacuum cap or funnel with the tab facing down and aligned with vacuum cap sidearm .If the alignment guide covers the sealing surface of the vacuum cap or funnel, it is upside down. Remove and change guide alignment guide orientation so that the sealing surfaces of the cap/funnel and base can be assembled in direct contact with one another .NOTE: The alignment guide is helpful in aligning the cap or funneland base, but it is not required.3. For continuous filtration using the vacuum cap , connect an appropriate length of PTFE tubing to thesidearm (inlet) of the cap.NOTE: Use the shortest length of tubing possible between the solvent container and the vacuum cap. Slowly push the tubing over the sidearm of the cap. If necessary, soften it first by immersing the end (approximately 1 centimeter [cm]) in hot water before pushing it over the sidearm.4. Place the Millisolve™ vacuum flask on a flat surface close to the vacuum source. Place the base on top of theflask.5. If using the stainless steel screen filter support, install it in the base, making sure that the supplied PTFE gas-ket is installed under the support.6. With smooth-tip forceps, center a 47 mm filter disc on thesupport surface.7. Without disturbing the filter, center the flange of the vacuum capor funnel on top of the base and lock the cap or funnel and basetogether with the spring clamp.8. Connect the free end of the vacuum tubing to the vacuum source.9. For continuous filtration (using the vacuum cap), pour the buffer/solvent to be filtered into a beaker,Erlenmeyer flask, or other suitable laboratory-grade container. Immerse the end of the PTFE tubing into the solutionto be filtered, making sure that the end reaches to the bottom of the container.For batch filtration (using the funnel), pour the solution to be filtered into the funnel.CAUTION: Make sure that the volume you want to filter does not exceed the 2 L capacity of the Millisolve™ flask.A 5 L flask (XX1614706) is available for larger volumes.10. Apply vacuum. Do not exceed 948 millibar (mbar) (28 in. Hg).For continuous filtration, the liquid will be drawn into the PTFE tubing, through the vacuum cap, through thefilter, and into the Millisolve™ flask.11. After all of the buffer/solvent has filtered, turn off the vacuum and disconnect the vacuum tubing from the vacuumsource.12. Remove the spring clamp, and lift the cap off of the base. Place the cap on a clean surface.13. With smooth-tip forceps, remove the filter from the base and discard.Using the Filtered Buffer/SolventThe Millisolve™ flask can be used as a solvent/buffer reservoir for theHigh Performance Liquid Chromatography (HPLC) or LC system. The Arrayconical bottom of the vacuum flask allows nearly the entire volume offiltered liquid to be utilized.1. Pass required lengths the LC solvent/buffer tubing through the 3-holestopper. If desired, attach a solvent filter/sparger to the ends of thetubing.2. Insert the 3-hole stopper into the flask.3. Connect the LC solvent/buffer tubing to the LC system and heliumsource.4. Pump the buffer through the HPLC or LC system.CleaningClean the system immediately after use. Depending on the solution filtered, you can flush the system with a filtered, volatile solvent (acetone or acetonitrile) or hot water, or disassemble and clean as follows:1. Rinse all components with hot tap water. Clean the porous glass frit of the filter support by back-flushing with tapwater.CAUTION: Do not clean with detergents, as they may introduce contaminants into the system.When cleaning the filter holder with the stainless steel screen filter support, the filter support drops outeasily when the base is inverted. Take care not to mislay or damage the PTFE gasket that lies under thefilter support.2. After rinsing thoroughly with tap water, rinse the components with laboratory-grade water.NOTE: Final rinse water can leave residues that affect critical tests. Use rinse water suitable for your application. 3. Allow components to air dry while disassembled.CAUTION: Do not wipe dry with paper or cloth, as this leaves fibers and lint on the surfaces and also generates electrostatic forces that can attract more dirt. To facilitate drying, flush with a volatile solvent such asacetone or acetonitrile.SpecificationsMaterialsMillisolve™ flask, vacuum cap, filter holder base, funnel ClampFunnel alignment guide3-hole stopper Borosilicate glass Anodized aluminum Silicone PolypropyleneFilter diameter47 mmEffective filter area Approximately 9.6 square centimeters (cm²) (1.5 in²) Funnel capacity300 mLVacuum flask capacity 2 LMaximum pressure948 mbar (28 in. Hg)Approximate dimensionsHeight with vacuum cap Height with funnel Vacuum flask diameter 45.7 cm (18.0 in.)53.3 cm (21.0 in.)12.5 cm (4.9 in.)Ordering InformationThis section lists catalogue numbers for the Millisolve™ Clarification System, replacement parts, and accessories. See the Technical Assistance section for contact information. You can purchase these products online at/products.Product DescriptionMillisolve™ Clarification System XX1614700 Replacement PartsVacuum cap XX1614701Vacuum base XX1514702Funnel, 300 mL XX1014704Clamp XX1014703Vacuum flask with conical bottom, 2 L, ground glass attachment XX1614705Funnel alignment guide ALIGNED05Filter forceps, stainless, smooth-tip XX6200006PPTFE tubing for solvent and buffer, 70 cm (27.6 in.)XT1200000 Accessories for Millisolve™ Clarification SystemBaseXX1514732 Vacuum base with stainless steel screen filter support and gasket,ground glass attachmentFunnelFunnel, 500 mL XX5014704Flasks with ground glass attachmentVacuum filtering flask, 4 L XX1014745Vacuum filtering flask, 1 L XX1514705Vacuum filtering flask with conical bottom, 5 L XX1614706Flask with stopper attachment for use as vacuum trapVacuum filtering flask, 1 L XX1014705Vacuum filtering flask, 1 L, threaded sidearm XX1514706PumpsChemical Duty Pump, 115 V, 60 Hz WP6111560Chemical Duty Pump, 100 V, 50/60 Hz WP6110060Chemical Duty Pump, 220 V, 50 Hz WP6122050FiltersMF-Millipore™ 0.22 µm hydrophilic MCE filter, Triton® surfactant-free, 47 mm, 100/pk GSTF04700Durapore® 0.22 µm hydrophilic PVDF filter, 47 mm, 100/pk GVWP04700Nylon 0.2 µm hydrophilic membrane filter, 47 mm, 100/pk GNWP04700Fluoropore™ 0.2 µm hydrophobic PTFE filter, 47 mm, 100/pk FGLP04700Omnipore™ 0.2 µm hydrophilic PTFE filter, 47 mm, 100/pk JGWP04700MF-Millipore™ 0.45 µm hydrophilic MCE filter, Triton® surfactant-free, 47 mm, 100/pk HATF04700Durapore® 0.45 µm hydrophilic PVDF filter, 47 mm, 100/pk HVLP04700Nylon 0.45 µm hydrophilic membrane filter, 47 mm, 100/pk HNWP04700Fluoropore™ 0.45 µm hydrophobic PTFE filter, 47 mm, 100/pk FHLP04700Fluoropore™ 0.45 µm hydrophobic PTFE filter, unbacked, 47 mm, 100/pk FHUP04700Omnipore™ 0.45 µm hydrophilic PTFE filter, 47 mm, 100/pk JHWP04700LCR 0.45 µm hydrophilic low extractables PTFE filter, 47 mm, 100/pk FHLC04700MiscellaneousMillex®-FA50 filter, 1.0 µm hydrophobic PTFE, 50 mm, 10/pk SLFA05010Tubing, 3/16 in. (4.8 mm) ID x 4.6 ft (140 cm), silicone with Luer adapter XX7100004NoticeThe information in this document is subject to change without notice and should not be construed as a commitment by EMD Millipore Corporation (“Millipore”) or an affiliate. Neither EMD Millipore Corporation nor any of its affiliates assumes responsibility for any errors that may appear in this document.T echnical AssistanceFor more information, contact the office nearest you. In the U.S., call 1-800-645-5476. Outside the U.S., go to our web site at /offices for up-to-date worldwide contact information. You can also visit the tech service page on our web site at /techservice.Standard WarrantyThe applicable warranty for the products listed in this publication may be found at /terms (“Conditions of Sale”).Millipore, Millex, and Durapore are registered trademarks of Merck KGaA, Darmstadt, Germany.The M initial, Millisolve, MF-Millipore, Omnipore, and Fluoropore are trademarks of Merck KGaA, Darmstadt, Germany.All trademarks of third parties are the property of their respective owners.© 2017 EMD Millipore Corporation. Billerica, MA, U.S.A. All rights reserved.20231184, Rev. 06/17。

Merck Millipore-液相芯片技术在肿瘤检测和研究中的应用

Milliplex/Luminex液相芯片技术在肿瘤检测和研究的应用前言目前，肿瘤标志物的研究与应用已成为肿瘤防治的重点和热点。

但当前肿瘤标志物检测能否达到早期诊断的效果？有无在人群中进行普查或筛查的价值？其临床意义如何解释？怎样规范与合理应用？尚存在诸多争议。

为此，中华医学会检验医学分会肿瘤标志物专家委员会，在2002年至2004年分别召开3次专家研讨会，起草制订了“肿瘤标志物临床检测的基本原则(建议稿) ”对以上问题进行了解释。

本文主要从以下几个方面进行介绍：●肿瘤标志物临床检测的基本原则●肿瘤标志物的检测指标●肿瘤标志物的高危人群检测项目●肿瘤血管的形成●慢性炎症与肿瘤微环境●趋化因子与肿瘤微环境●液相芯片技术原理●Milliplex肿瘤微环境相关试剂盒介绍●肿瘤微环境与肿瘤标记应用的相关文献一、肿瘤标志物临床检测的基本原则肿瘤标志物( TM) 是指在恶性肿瘤发生和增殖过程中，由肿瘤细胞的基因表达而合成分泌的或是由机体对肿瘤反应而异常产生和/ 或升高的，反映肿瘤存在和生长的一类物质，包括蛋白质、激素、酶(同工酶) 、多胺及癌基因产物等。

TM 存在于病人的血液、体液、细胞或组织中,可用生物化学、免疫学及分子生物学等方法测定，且对肿瘤的辅助诊断、鉴别诊断、观察疗效、监测复发以及预后评价具有一定的价值。

TM在肿瘤监测中的价值：TM的主要临床应用价值是判断治疗肿瘤治疗疗效和复发监测。

临床可通过对肿瘤患者治疗前后及随访中TM浓度变化的监测，了解肿瘤治疗是否有效，并判断其预后，为进一步治疗提供参考依据。

为确定何种TM适用于对肿瘤患者进行治疗监测，在患者治疗前应做相关TM检测。

TM浓度变化对肿瘤的疗效判断价值：恶性肿瘤治疗后TM浓度的变化与疗效之间有一定的相关性。

治疗前TM浓度变化,常有三种类型：①TM浓度下降到参考范围，提示肿瘤治疗有效。

②TM浓度下降但仍持续在参考范围以上，提示有肿瘤残留和/ 或肿瘤转移。

流式细胞仪使用方案

常见荧光素波长分布示意图
二、同型对照的选择：
流式细胞实验和其他抗体相关实验有点不同的就是需要选择同型对照。同型对照，是用于消除由于抗体非特异性结合到细胞表面而产生的背景染色，相当于实验的阴性对照。同型对照选择与标记抗体同种属来源，同亚型，同荧光标记的抗体。比如：抗人的CD3的PE标记的抗体，小鼠的IgG2a。同型对照选择PE标记的小鼠的IgG2a。
10固定30min4度冰箱过夜或20度长时保存二染色技术1选取荧光素2样品染色步骤pbs洗细胞12次加荧光单抗暗处孵育1530min如果必需加荧光二抗暗处孵育1530minpbs洗细胞一次过300目尼龙网上机检测3对照的设置阴性对照同型对照见附录三流式细胞数据处理与分析数据的显示通
流式细胞仪使用方案
2流式抗体荧光标记的方式包括直接标记和间接标记两种。在流式实验过程中，尽量减少实验工序和过程，以保证实验的真实和准确性。因此在条件允许的范围内，建议尽量用直接标记的抗体进行实验而不去做间接标记。
3流式抗体荧光标记的选择：如果实验中检测单一指标：不同荧光标记在不同的仪器上强度不同。FACS Calibur仪器为例：PE＞APC＞PE-Cy5＞PerCP＞FITC＞PerCP-Cy5.5，通常来说，PE最强，适用于弱表达抗原。FITC强度较弱，适用于强表达抗原，使用范围比较广。用户需根据检测的目标蛋白进行具体选择。如果同时检测多个指标：确认流式细胞仪能检测多少个通道：流式抗体每个通道只能选择1种荧光素。各个通道之间的荧光素可以随意搭配。如：实验者同时检测三个指标，可以在图1中绿色、黄色和红色三个通道中各选一个适当的荧光素标记，FITC、PE和PE-cy5。切忌所有指标选择同一个通道的荧光标记，以防止荧光的重叠和相互干扰，影响最后结果。因此，流式细胞仪的通道越多，同一份样本能同时做的表面/胞内标志就越多。常用荧光标记包括FITC,PE, PEcy5, PEcy5.5, APC等。

MAGPIX操作手册简介2

MAGPIX操作指南Merck Millipore 生命科学部开始使用您的MAGPIX系统目录:• MAGPIX系统简介 (3)• MAGPIX安装快速指南 (7)• MAGPIX操作快速指南 (16)MAGPIX技术简介一、Luminex液相芯片检测系统介绍液相芯片技术是20世纪90年代后期发展起来的被喻为后基因组时代的芯片技术。

液相芯片技术将流式检测技术与芯片技术有机地结合在一起，一方面大大延伸了流式检测平台，以微球体代替细胞作为反应载体，这个相当开放的反应体系可进行蛋白、核酸等等生物大分子的检测，不仅从细胞水平深入到分子水平，其检测范围也得到了前所未有的扩展；另一方面也使芯片技术取得重大的突破：在保持高通量检测的同时，将反应体系由液相－固相反应改变为接近生物系统内部环境的完全液相反应体系，对于确保建立在正确的高级结构之上的真实的蛋白质相互作用尤为关键。

迄今为止不到十年的时间，全球已有数百套基于此项技术（xMAP：flexible Multi-Analyte Profiling）的检测平台，用于蛋白质，免疫学等等领域的研究，临床诊断试剂的开发方面也发展迅速，自身免疫鉴别诊断试剂等已通过了FDA认证。

二、Liminex液相芯片检测系统技术原理Luminex液相芯片检测系统整合微球双荧光标记和液流分散激光自动检测技术，自动实现核酸、酶、受体、抗体、抗原、小分子有机物等多通道高通量分析。

核心技术为特制的100种不同色标编码的5.6um 塑料小球或6.5um的磁珠。

每一种编码的小球标记一种可以捕获相应目标分子的配体，任选几种或多至100 种标记好的小球混合后与样品中待检目标分子作用，由于每种小球标上不同的探针分子，从而可以对样本中多大100中的不同种目标分子进行同步检测。

三、Liminex液相芯片检测系统重要组成部分1. 微球通过两种荧光染料染色得到，调节其之间的荧光比例可以获得100种不同颜色的微球（5.6um聚丙烯微球或 6.5um磁珠），每种颜色的微球可以携带一种生物探针。

Merck MilliporeSigma AFS 8 16 24 水纯化系统说明书

The life science business of Merck operates as MilliporeSigma in the U.S. and Canada.AFS ®8 / 16 / 24Water Purification SystemsCost-effective solutions for clinical analyzers with pure water needs up to 480 L dailycost-effective solutionsFor clinical analyzers with pure water needs up to 480 L dailyA reliable and constant source of clinical laboratory reagent water (CLRW)that complies with the CLSI® guideline*Complementary water purification techniques, including complete pretreatment, advanced reverse osmosis (RO), exposure to a bactericidal UV lamp, and final 0.22 μm filtration, provide consistent water quality meeting CLRW standards.A cost-effective solution Optimized lifetimes for pretreatment and polishingpacks, as well as low feed water use make AFS® systemsa cost-effective choice.A system that fulfills accreditation needs (e.g., CAP 15189SM accreditation to the ISO® 15189:2007 standard)AFS® systems have full monitoring and automatic water quality archiving capabilities. Up to six months of information can be stored for reliable traceability, and Millitrack® software provides easy access to data.A robust system requiring little maintenance An intuitive icon-based system display facilitates maintenance, and a new ergonomic pack locking system makes pack changes easier than ever. Reduced maintenance saves time and also means less analyzer downtime.Professional, rapid service Our systems are backed by a responsive, professionalservice organization providing rapid intervention.Milli-Q® Service Plans offer a range of support, includingpreventive maintenance visits.An adaptable configuration that makes optimaluse of lab space AFS® systems have a small footprint, allowing installation wherever it’s convenient: on the wall, on or under the bench, or on a cart. A wide range of reservoirs handles a variety of water storage needs. Add production capacity by installing a RiOs™ system in duplex.Confidence in your water purification system supplier As one of the top three R&D investors in the Life Science Tools industry and with more than 50 years of experience in water purification systems manufacturing, we are a partner you can count on.*Clinical and Laboratory Standards Institute, Inc. (CLSI) guideline: “Preparation and Testing of Reagent Water in the Clinical Laboratory; Approved Guideline – Fourth Edition” (CLSI C3-A4)273Constant- and reliable-quality clinical laboratory reagent water (CLRW)AFS ® Systems’ Water Purification Pathway1. Feed Water2. Inlet Value3. Progard ® Pretreatment Pack4. Pressure Regulator (RO)5. RO Pump6. Feed Water Conductivity Cell7. Sanitization Port8. RO Membrane9. RO Pressure Sensor 10. Flush Valve11. Reject Water Recovery Loop 12. Reject13. Permeate Conductivity Cell 14. Rinsing Valve 15. Reservoir16. Distribution Pump17. Q-Gard ® Polishing Pack 18. Distribution Resistivity Cell 19. UV Lamp (254 nm)20. Distribution PressureRegulator21. Distribution Pressure Sensor 22. 0.22 μm Final Filter 23. Sampling Valve 24. AnalyzerCombined purification technologies for the best resultsIn biomedical laboratories, pure water is an essential reagent that is necessary for delivering consistent, high quality results, maximizing productivity and improving patient outcomes. Different types of contaminants may interfere with biomedical lab analyses and alter test results, which is why our AFS ® systems employ a combination of purification technologies.Water produced by AFS ® systems has consistent quality meeting Clinical and Laboratory Standards Institute, Inc. (CLSI ®) CLRW standards:• Resistivity > 10 MΩ·cm @ 25°C • Bacteria level < 10 cfu/mL• Total Organic Carbon (TOC) < 500 ppb • 0.22 μm filtrationIn the AFS ® system water purification sequence, potable tap water is first treated with a Progard ® cartridge, and then purified by advanced reverse osmosis to remove up to 99% of contaminants. Regardless of seasonal temperature changes, the AFS ® system produces a constant flow of RO water, ensuring there is always enough pure water.Optimized control over water quality for low bacterial levelsThe RO water is stored in a reservoir. To ensure optimal water quality, stored water is automatically recirculated through a Q-Gard ® polishing pack and a built-in 254 nm bactericidal UV lamp. As a last step, the pure water is sent through a sterilizing-grade filter before entering the analyzer. This helps to limit downtime resulting from analyzer decontamination.Depending on your needs, we offer either a 0.22 μm Opticap ® filter to remove particles and bacteria, or a BioPak ® C clinical ultrafiltration cartridge that will remove bacterial by-products for alkaline phosphatase-free water.4A water purification system fulfilling accreditation needsToday, recent significant improvements in quality management systems are leading biomedical laboratories more and more frequently to seek accreditation to the ISO ® 15189:2007 standard, which is supported by CAP 15189SM accreditation, or by “The Key to Quality” workbook from the CLSI ®.*As the most commonly used fluidic reagentonboard an analyzer, water is a critical reagent, and its quality must be documented. AFS ® systems have full monitoring and automatic water quality archiving capabilities. Up to six months of information is stored for easy and reliable traceability. Optional activation of Millitrack ® software provides quick access to thisdata, as well as enhanced data management control and remote access capabilities.AFS ® systems check and also archive relevantparameters, helping to control proper operation of the system. Risk management and reduced maintenance are achieved thanks to several adjustable set points that can trigger an alarm in case of deviation. Values such as feed pressure, feed water quality, RO pressure, RO water quality, and RO membrane efficiency (% ion rejection), etc., can be displayed on the system’s LCD display.*International Organization for Standardization (ISO ); College of American Pathologists (CAP); Clinical and Laboratory Standards Institute, Inc. (CLSI )A cost-effective solution to your clinical water purification needsBudget-conscious users will appreciate AFS ® systems for their optimized running costs. Advanced AFS ® system design as well as software with and increase cartridge lifetimes:• The single Progard ® pretreatment pack incorporates severalfree chlorine, and scaling.® pack.condition.of the Q-Gard ® pack.5A low-maintenance, robust systemWith only the Progard ® pretreatment pack and Q-Gard ® polishing pack to change, AFS ® systems provide a low-maintenance solution to your clinical laboratory water purification needs — saving you time and also reducing analyzer downtime.To help make pack changes even easier, AFS ® systems have a new ergonomic pack locking system. Just pull up on the locking handle to remove the exhausted pack, position the replacement pack in the cabinet, and push down on the handle to lock the new pack in place — it’s as simple as that!In addition, RFID technology prevents insertion of an incorrect purification cartridge in AFS ® systems, and also ensures traceability of pack use and replacement.Essential information at a glanceAFS ® systems have been designed for easy, effortless operation. Intuitive controls on the system cabinet simplify use — you see just the information you need, such as product water quality and reservoir water level. Whennecessary, icons and the system’s backlit LCD screen change color to visually inform users of any actions that should be performed.• Blue display: normal operation • Yellow display: maintenance needed • Red display: urgent action requiredWhen there has been no user interaction with the screen for 15 minutes, and there is no alert or alarm, the system’s screen saver (“ECO mode”) will be activated automatically.Additional information on system operation and maintenance is provided by the Quick Reference Guide and User Manual stored on the water production unit.Professional, rapid serviceAFS ® systems are backed by a responsive, professional service organization providing rapid intervention. When Millitrack ® software is activated, this can further facilitate a quick diagnostic from the AFS ® system dashboard. An optional emergency backup function can be installed to provide AFS ® customers with temporary coverage until their system is serviced.Milli-Q ® Service Plans offer a range of support, from a single annual checkup to full system coverage. Our certified Field Service Engineers provide expert, professional support for the installation and maintenance of your AFS ® water purification systems, and our technical hotline support experts are available to investigate, diagnose and solve customer issues.6An adaptable configurationConfidence in your water purification system supplieras water contaminants.AFS® systems are manufactured in an ISO® 9001-and ISO® 14001-registered site. Additionally, toreduce environmental impact, all AFS® systems(WEEE) directives.Optimal use of lab spaceWith their small footprint, AFS® systems can be placed whereverit’s convenient — on the wall, on or under the bench, or on a cart.A wide range of storage reservoirsSelect from a wide range of high quality polyethylene reservoirs (10 – 350 L)to match your water usage. Our reservoirs benefit from a number of features that maintain consistent purity of stored water and provide effective protection against airborne contaminants:• A reservoir vent filter protects water from particulates, bacteriaand dissolved CO2.• An aseptic overflow function maintains water quality by avoidingretro-contamination from the drain.• The conical reservoir base allows complete draining, and facilitates rinsing during sanitization, while the smooth interior limits biofilm formation.Systems that can be adapted to your needsAFS® systems are available with numerous options including: an Automatic Sanitization Module (ASM) for further bactericidal protection of stored water; sanitary sampling valve; sanitization kit for full decontamination of the entire system and reservoir; and water sensor.To add production capacity to your AFS® system, a RiOs™ system can be installed in duplex to provide up to 48 L/hour of pure water.AFS® system specificationsSystem PerformanceResistivity > 15 MΩ.cm @ 25°CTotal Organic Carbon (TOC)Typically < 30 ppbBacteria< 1 cfu/mLDissolved silica< 0.05 mg/LDispensing flow rate to analyzer Up to 2 L/minDispensing pressure to analyzer0.9 –3 bar (adjustable)Production flow rate to reservoir8 L/h (AFS 8), 16 L/h (AFS 16), 24 L/h (AFS 24)System InformationDimensions (H x W x D) 585 x 268 x 426 mm (23 x 10.6 x 16.8 in)Net weight (shipping box)< 23.1 kg (50.9 lb)Operating weight< 28 kg (61.7 lb)Voltage100 – 240 VACFrequency50 – 60 HzPower consumption200 W or 250 VAFeed Water Quality RequirementsPressure 1 – 6 barFlow rate> 5 L/min at 2 barTap Water Connection½ in. Gaz MType PotableTemperature 5 –35°CConductivity100 – 2000 μS/cm at 25°CpH 4 –10Langelier Saturation Index (LSI)< 0.3Merck KGaA Frankfurter Strasse 250 64293 Darmstadt Germany© 2018 Merck KGaA, Darmstadt, Germany and/or its affiliates. All Rights Reserved. Merck, the vibrant M, Milli-Q, AFS, BioPak C, Millitrack, Opticap, Progard, Q-Gard and RiOs are trademarks of Merck KGaA, Darmstadt, Germany or its affiliates. All other trademarks are the property of their respective owners. Detailed information on trademarks is available via publicly accessible resources.PB4337EN00 Ver. 2.02017 - 0124601/2018To Place an Order or Receive Technical AssistanceIn Europe, please call Customer Service: France: 0825 045 645Germany************Italy: 848 845 645Spain: 901 516 645 Option 1 Switzerland: 0848 645 645 UnitedKingdom************For other countries across Europe,please call: +44 (0) 115 943 0840For other countries across Europeand the world, please visit: /officesFor Technical Service visit: /techservice。

MILLIPORE凋亡试剂盒操作流程

自己翻译的，如有出错，请多指教该试剂盒的优点在于可以迅速得到结果，操作简单，并且结果清晰，资金允许的话强烈推荐〜需要准备的试剂1蛋白酶K(20卩g/m石蜡切片适用2.正丁醇3.酒精：乙酸2：14.1 %多聚甲醛-PBS (无甲醇甲醛固定的冰冻切片或细胞)。

5.10(V:V) 中性福尔马林(石蜡包埋固定前)6.PBS (50mM 磷酸钠，PH7.4 200mM NACL)7.30%过氧化氢8.Triton X-100 10%9.柠檬酸缓冲液(PH=6) 294g+100ml H2OC6H5NA3O7.2H2O10.P BS (PH=7.4) NA2 HPO4 55gNAH2 PO4 13.5gNACL 117 gH2O 1L试剂的准备与安装下面推荐的试剂量，足够充分覆盖组织浓缩的TDT酶在平衡缓冲液中保留其活性。

在使用前应必须与反应缓冲稀释后才能使用。

混合试剂中70%反应液30%的TDT酶。

在准备时用新的微离心管。

Vol/5 cm2ReagentEquilibration Buffer 平衡缓冲液13 ^L65 ^LWorking Strength TdT 工作强度转移酶11 yL55 yLAnti-Digoxigenin- Peroxidase 过氧化物酶13 yL65 yLDAB Peroxidase Substrate DAE过氧化物底物15 yL75 yL77 y L 33 yL110 yLReaction Buffer (90417) TdT Enzyme (90418) 反应液TDT酶Total用漩涡式振荡，该试剂在冰上存放时间不超过6H, 这能足够对付5CM 2个标本196蛋白消化酶或蛋白酶K200 g/mL蛋白酶K在20卩g/m稀释。

在200 ®/mL的蛋白酶K 3.9ML到35ML PBS以给予足够稀释的量。

直接加样，将30%的双氧水用PBS稀释为3%浓度。

可以在30MLPBS 中加入4ML 30% 的H2O2。

胆汁中外泌体及外泌体RNA提取方法比较

胆汁中外泌体及外泌体RNA提取方法比较王有礼; 谭明娟; 葛贤秀【期刊名称】《《检验医学与临床》》【年(卷),期】2019(016)023【总页数】5页(P3405-3409)【关键词】胆汁; 外泌体; 核糖核酸; 超速离心; 聚合沉淀【作者】王有礼; 谭明娟; 葛贤秀【作者单位】南京医科大学附属南京医院/南京市第一医院检验科江苏南京210006; 南京医科大学第二附属医院消化科江苏南京210011【正文语种】中文【中图分类】Q522外泌体是细胞内多胞体与质膜融合后分泌到细胞外环境中的一种膜性囊泡。

外泌体来源于晚期核内体，由活细胞分泌而来，几乎所有的细胞都能分泌，广泛存在于各种体液中。

研究发现，外泌体可通过囊泡运输方式，在体内充当细胞间通讯使者角色[1]；在免疫应答、凋亡、血管生成、炎症及肿瘤发生、发展等过程中均有作用[2-3]。

目前，外泌体分离和纯化方法尚不统一，对临床转化造成一定困难。

胆汁是胆管上皮细胞赖以生存的微环境，特别是其中所含外泌体中蕴含丰富的胆道疾病信息，是应用价值很高的液体活检标本。

关于对人胆汁进行分离纯化的研究较少，大多采用超高速差速离心法[4- 5],但该方法具有操作耗时、所得标本量少等缺点。

因此，寻找灵敏度高、简便快捷的提取方法至关重要。

本文比较超速离心法和聚合沉淀法提取人胆汁外泌体和外泌体RNA的效果，现报道如下。

1 资料与方法1.1 标本收集 2017年6月至2018年12月，收集南京医科大学第二附属医院消化内科因胆总管结石行内窥镜逆行胆囊-胰腺造影术(ERCP)的患者15例，标本为ERCP术中插管后造影前抽取得到的胆汁，每例患者胆汁收集量大于10 mL。

所有病例均经过影像学检查确认。

所有患者术前均空腹，手术均由熟练的操作者实施，获取的胆汁提取后均立即储存于-80 ℃冰箱待用。

本研究得到南京医科大学第二附属医院伦理委员会批准，纳入研究的患者均于术前签署知情同意书。

1.2 仪器与试剂超速离心机：Optima L-100 XP Ultracentrifuge(Beckman Coulter，美国)，与之适配的Beckman超速离心管(50 mL，Beckman Coulter，美国)；外泌体提取试剂： Total Exosome Isolation Reagent (from other body fluids) (Invitrogen，美国)；0.22 μm滤器(Merck Millipore，美国)；透射电镜：Tecnai G2 Spirit Bio TWIN(FEI，美国)；粒径检测仪器：ZETASIZER Nano series-Nano-ZS (Malvern,美国)；流式细胞术：Accuri C6(BD Biosciences，美国)；CD63和CD81抗体(BD Biosciences，美国)；RNA提取试剂：Trizol (Invitrogen，美国)；反转录试剂盒：Taqman MicroRNA Reverse Transcription KIT，引物Taqman microRNA assays(applied biosystems，美国)；实时荧光定量PCR试剂盒：Taqman Universal Master Mix Ⅱ，noUNG(applied biosystems，美国)。

Merck Millipore Direct-Q 水纯化系统用户指南说明书

Tap to pure and ultrapure water ― with easy and convenient dispense!Your water purification needs Our solution: the Direct-Q® range of water purification systemsHigh quality water produced directly from tap water Direct-Q® systems deliver both pure and ultra-pure water directly from tap for use with a wide variety of applications in your lab.Ultrapure water easily accessible wherever you need it in your lab With the Direct-Q® range of water purification systems, you benefit from a choice of ultrapure water dispensing possibilities. The innovative, space-saving Remote dispenser offers you water delivery solutions to best fit the way you work, with easy and convenient remote delivery up to two meters away from your water production unit.Compact all-in-one system for the most efficient use of your lab space The Direct-Q® range includes versatile solutions that can be bench-integrated or bench- / wall-installed.A choice of adequate storage volumes Direct-Q® systems come either with an integra-ted 6-liter reservoir or an external reservoir (30 l or 60 l).Flow rates adapted to your daily usage Systems in the Direct-Q® range provide 3, 5 or 8 liters of pure water per hour, covering lab needs of 50,100 and 150 l of pure water per day.Instant flow rates to match your needs Direct-Q® systems dispense up to 30 l ofultrapure water per hour.High quality water to meet the requirements of your most critical applications Options such as a UV lamp and a range of Application Pak point-of-use polishers are available to fine-tune your ultrapure water.Easily accessible information on system operation The user-friendly display provides system status at a glance; the concise Quick Reference Guide is a handy guide for daily operation.Simple, low-level self-maintenance All-in-one SmartPak®cartridges enable easy and rapid replacement.Direct-Q® Systems Water Purification Pathway Choose the solution that’s right for youEasy installationThe Direct-Q® range of systems requires no special installation. You can easily set the system up yourself: just connect the system to your tap water supply, plug it in, and insert the SmartPak® purification cartridges. Then, if your configuration includes the Remote dispenser or a 30- or 60-liter reservoir, follow the simple setup procedures ― and your system is ready to use!Optimized lab spaceThe compact, all-in-one Direct-Q® water purification unit lets you locate it nearly anywhere in your laboratory, either on or under the benchtop, or wall-installed.**Direct-Q® 5 and Direct-Q® 8 systems are designed for use with a 30 l or 60 l external reservoir.Organic-sensitive applicationsDirect-Q® systems are also available with a built-in 185/254 nm UV lamp to reduce the level of organics for critical applications. Water with low TOC provides important benefits to HPLC users such as higher sensiti-vity and longer column lifetime. The same UV lamp also destroys bacteria.Application Pak point-of-use polishersMerck Millipore’s range of Application Pak polishers makes it possible to fine-tune your ultrapure water quality to match your research needs. Are your applications sensitive to bacteria, particulates, pyrogens, nucleases, endocrine disruptors or Volatile Organic Compounds? If so, just choose the appropriate final polisher from our range of Application Paks to provide optimal water quality for your requirements.Please see /labwater for more information.Both pure and ultrapure waterThe Direct-Q® range of systems provides you with a convenient and flexible solution for your pure and ultrapure water needs, directly from potable tap water. You’ll have access to ultrapure water for your critical applications, and pure water for less critical applications such as general glassware washing or final rinsing ― from the same system!The high quality ultrapure water produced by Direct-Q® systems is suitable for applications such as production of mobile phase for chromatographic separations; preparation of blanks and standard solutions for spectrophotometry, spectroscopy or other analytical techniques; and preparation of buffers for biochemical experiments.Flow rates to match your requirementsChoose the solution that best meets your lab’s requirements with a Direct-Q®system that provides 3, 5, or 8 liters of pure water per hour, and more than 0.5 l of ultrapure water per minute (> 30 l per hour). When not in use, the Direct-Q® system automatically recirculates water in order to maintain high water quality,so that you do not have to wait when sourcing ultrapure water. For your conve-nience, and to save time, you can also set the system to automatically deliver your selected volume of ultrapure water on demand.Choice of storage volumesWith its built-in reservoir, the Direct-Q® 3 system can store 6 l of reverse osmosis (RO) water, while Direct-Q® 5 and 8 models are designed for use with a 30- or 60-liter reservoir. Just choose the storage volume that best meets yourdaily water volume needs.Fine-tune your water qualityVersatile remote dispenserDesigned to fit perfectly into your lab environment, the versatile Remote dispenser can be placed up to two meters from your Direct-Q® water purification unit. Select the free-standing or wall-installed model according to your needs ― their ergonomy will make either one a welcome addition to your lab,giving you the freedom to focus on your research, while dispensing ultrapure water exactly where you need it. Alternatively, Direct-Q® systems are also available with an integrated dispenser for use on the benchtop.The design of the bench-installed Direct-Q® systems is also adapted to the height and shape of common laboratory glassware.Stay focused on your workMerck Millipore offers more than waterJust the information you needThe intuitive color graphic display shows key system parameters at a glance,enabling easy water quality and maintenance warnings monitoring; the screen rotates for easy viewing wherever thesystem is located. A short Quick Reference Guide is conveniently located in the base of the system for immediate information; the complete User Manual can be stored at the back of the water purification unit.User-friendly maintenanceThe all-in-one SmartPak® purification cartridges are easily replaced in just a few minutes.Watercare Pact Service portfolioTo optimize the performance and lifetime of your water purification system,Merck Millipore offers a complete portfolio of Service plans ranging from a single annual checkup to a full system cover. For more information, please check with your Merck Millipore applications specialist or visit our website:/labwaterSpecificationsAvailable System ConfigurationsWater Purification Systems* 30 l or 60 l depending on daily water usageFor more information, please visit our website:/directq358Millipore, Direct-Q®, SmartPak®, Millipak® and BioPak® are registered trademarks of Millipore Corporation. Merck Millipore and the M mark are trademarks of Merck KGaA.Lit. No. PB2040EN00© 2011 Millipore Corporation, Billerica, MA, U.S.A. All rights reserved.。

成像流式细胞仪原理及应用

二代流式技术产品-成像流式细胞分析邱又彬Merck&Millipore旗下品牌Amnis于近期推出了新一代高速细胞成像系统ImageStreamX Mark II。

这是第三代ImageStream成像流式细胞仪，具有无与伦比的细胞分析能力。

显微镜可提供详细的细胞图像和形态信息，是研究细胞功能的重要工具。

然而，显微图像的解释却是主观、定性且费力的。

流式细胞仪擅长定量的表型分析，可产生统计学上可靠的结果，不过，流式细胞仪却缺乏成像能力，因此无法了解亚细胞定位。

ImageStream系列开创性地将流式细胞检测与荧光显微成像结合于一体，既能提供细胞群的统计数据，又可以获得单个细胞的图像，从而提供了细胞形态学、细胞结构和亚细胞信号分布的完整信息。

ImageStreamX Mark II能实时捕获每个流动细胞最多12幅高分辨率图像，检测速率可达5000细胞/秒，并具有更强荧光灵敏度。

ImageStreamX Mark II的这些功能可以对细胞形态、荧光探针的强度和定位进行检测，进而为科学家提供广泛的图像分析应用，包括细胞间相互作用、吞噬、凋亡和自噬、核易位、形态变化等。

ImageStreamX Mark II的特征如下：•速度更快：Mark II对进样速度进行了提升，每秒可以分析多达5000个细胞，简单易用的补偿向导可以指导您轻松完成多色补偿运算。

•操作更简单：全新而直观的用户界面提供了每一个细胞的图像及实时绘图相关的图形化控件。

•样品适应性更强：Mark II可选配7个激光；样品容量20-200 μl，增加了实验的灵活性，适用于多用户实验室。

•样品利用率更高：Mark II将样品利用率提高到了95%，更适用于稀有的细胞样品，而且未使用的样品也可回收用于进一步分析。

从2005年开始，Amnis量化成像流式分析仪被广泛应用于各个研究领域。

其中超过300篇的文章发表在高水平的同行评议杂志（peer-reviewed journal）上。

Merck Millipore MAS-100 微生物空气监测系统说明书

Improve your viable air sampling with the MAS-100®The most complete and accurateline of microbial air monitoring systems available!How the MAS-100® worksAll MAS-100 ® Microbial Air Monitor- ing Systems are sieve impaction sys- tems based on the Anderson impac- tion principle, accepted and proven worldwide. Air is aspirated through a perforated lid. A radial fan, controlled by a flow sensor, accurately regulates air flow real time at 100 litres/min. The air is impacted onto the surface of growth media in a 90 – 100 mm Petri dish or 55 – 60 mm Contact Plate. MAS-100 ® Microbial Air Samplers comply with the guidelines as specified in new ISO 14698 part1 and part 2.Sample collectionaccuracyThe MAS-100 ® Microbial AirMonitoring Systems provide the mostaccurate sample collection available.The MAS-100 ® systems (except theMAS-100 Eco®) utilize a state-of-the-art mass flow sensor to ensure aconstant air flow rate of 100 litres/min.The MAS-100 VF® uses a state-of-the-art electronic flow control to ensurethe same constant flow rate. The massflow sensor continuously regulatesthe air flow rate during samplecollection. This allows the MAS-100 ®to automatically adjust for differencesin fill volume of Petri dishes, changesin air density and differencesbetween individual perforated lids.ICR* andICRplus Settle PlatesThe ICR Settle Plates areproduced under aseptic conditions,gamma-irradiated and triple-bagged,making them optimal for passive airsampling in isolators and cleanrooms.Furthermore, they are also suitablefor personnel monitoring, as wellas active air monitoring withMAS-100® microbial Air Samplers.* Isolators and Clean Rooms2Table of Contents04The MAS-100 Iso NT®06The MAS-100 Iso MH®08The MAS-100 VF®10The MAS-100 NT®11The MAS-100 NT® Ex 12The MAS-100 Eco®13The MAS-100 CG Ex®14The DA-100 NT®15Services & Support16ICR & ICRplus Settle Plates 18Ordering Information34100 The MAS-100 Iso NT ®For IsolatorsThe MAS-100 Iso NT® is specifically designed for use in aseptic production and sterility testing isolators. It is available in an IP54 version for increased flexibility. The MAS-100 Iso NT® is produced by GAMP 4 specifications and is compliant with guidelines as specified in ISO 14698 part1 and part 2. The MAS-100 Iso NT® uses an innovativedouble valve system which enables the sampling heads and unit to be integrated into the decontamination process of the isolator or cleanroom. The valve system allows the vaporized hydrogen peroxide(VHP), to run through the sampling head and internal flow path without damage to the instrument.5Technical specifications for MAS-100 Iso NT®The system operates with an integrated mass flow sensor and uses 90 – 100 mm Petri dishes. The sampling head mounts inside the isolator with the instrument portion on the outside. The MAS-100 Iso NT® utilizes a flexible communications packageincluding Ethernet, Profibus and 9 digital inputs/outputs. The MAS-100 Iso NT® allows for remote activation via a computer or the PLS controlled on the isolator.The new M AS-100 Iso NT® shares the same samplingaccuracy and features as the rest of the MAS-100® Family.For further information about our workshops and onsiteservices please have a look at page 15.6100 Decontaminant exaustThe MAS-100 Iso MH ®Innovation pure by Merck MilliporeThe MAS-100 Iso MH® is used to control the microbio- logical contamination of the air in isolators. A unique security concept permits the installation of the sampling heads for standard 90 – 100 mm Petri dishes at thecritical control points. All electronic and moving parts are outside the critical zone. The MAS-100 Iso MH® has an additional internal pump with flow control for automatic disinfection of the sampling head and the aspiration tube.Air exaustup to 1 meter7An innovative double valve system enables the sampling heads to be integrated into the decontamination process of the isolator. Each unit has an air- and a sterilisation pump and works fully autonomous.The MAS-100 Iso MH® is built according to GAMP 5 and corresponds to the ISO 14698 standard. It is a completly validated system designed for the highest demands.Technical Specifications for MAS-100 Iso MH®Filter (optional use)up to 10 meters8100 The MAS-100 VF ®The next generation of Air SamplersLike all MAS-100® systems, the MAS-100 VF® samples at a flow rate of 100 litres per minute. The handle enables the sampler to be mounted onto an optional tripod, thereby allowing testing at different angles for fixed applications. The perforated lid is the same as on the standard MAS-100® sampler.The new MAS-100 VF® active Air Sampler was especially developed for controlled environments. Regulatory expectations require air monitoring in manufacturing facilities because microbial contamination mayinfluence quality and reduce shelf life. The MAS-100 VF® uses 90 – 100 mm standard Petri dishes, is easy to handle and compact, while it is ideal for monitoring the quality of your environment. Its electronic speed control maintains an accurate flow rate.Technical Specifications-100 VF®The simple user menu is easily accessed and operatedby a single touch slide control. Sampling volumesare programmable from 1 – 1,000 litres, with 5 presetvolumes to assure reproducible results.910100 The MAS-100 NT ®The industry standard for viable air samplingThe MAS-100 NT ® and MAS-100 NT® Ex portable micro-bial airsamplers are the industry standard for use in critical environments. These compact yet sophisticated devices are the preferred choice for those demanding the highest quality in microbial air monitoring. The MAS-100 NT® systems feature a new 300-hole perfo-rated lid for increased collection efficiency and i mpaction speed. Both systems utilize standard 90 – 100 mm agar plates or can be adapted to fit 55 – 60 mm contact plates allowing for a low consumable cost and greater flexibility. Sampling at 100 litres per minute, thesesystems have the highest airflow accuracy available at ± 2.5 %, compared to others that can be as high as ±10 %. The integrated flow s ensor allows the user to freely interchange the perforated lids without affecting the accuracy or the calibration of the unit. Sampling volumes are also easily configurable b etween 1 and 2,000 litres. The units have an impact v elocity of 19.6 meters per second equivalent to Anderson 6 and isokinetic flow rate that will not produce turbulence in a laminar flow environment. The new SQS function will allow for smaller sampling volumes over longer periods of time, up to 50 sequences over 24 hours.The new MAS-100 NT® systems are controlled using a new menu driven, larger illuminated display allowing for quicker navigation. A programmable start delay of up to 60 minutes allows for personnel to be out of the sampling area when the sampling starts and a newaudible alarm indicating the interruption of a sampling11The MAS-100 NT ® ExExplosion prooffor use in explosion proof areas. The MAS-100 NT® Ex has received ATEx Conformity and can be used in zone 2 and gas groups 11A,11B and 11C in temperature classes T1 to T4.cycle. The MAS-100 NT® is powered by a newLithium ion rechargeable battery with an intelligent charging program that assures long battery life without routine discharging. When fully charged, the battery pack provides 7 hours of continuous operation or about 42,000 litres of total volume. The MAS-100 NT® Microbial Air Sampler also features new USB data communication port. This allows for easy download of software upgrades and easy communication with database programs. The improved communications provides an interface to the new DA-100 NT® calibration standard for fully automated calibration. The MAS-100 NT® and MAS-100 NT® Ex are the first microbial air sampling systems with automated calibration, assuring absolute accuracy.The MAS-100 NT® Ex shares all of the same functions of the MAS-100 NT® system but is specially designedTechnical specifications for MAS-100 NT® and MAS-100 NT® ExRTC (Real Time Clock) battery; good for approx. 10 years Hot-wire anemometer, numerical control, Temperature and Pressure -100 NT® Ex12Technical specifications for MAS-100 Eco®100 The MAS-100 Eco ®The MAS-100 Eco® offers an economical alternative to our highly acclaimed MAS-100® Microbial Air Samplers. This smaller, lightweight unit is ideal for applications in or out of the cleanroom. The MAS-100® Eco offers manyof the same features found on the standard MAS-100® Air Samplers, without the mass flow sensor. Like all MAS-100® systems, the MAS-100 Eco® samples at a flow rate of 100 litres per minute and uses standard90 – 100 mm Petri dishes, providing a low operating cost. The handle doubles as a stand to allow different angles for testing and an optional tripod adapter attaches to the handle for fixed applications. The simple user menu is easily accessed and controlled using single touch “yes ” or “no ” responses. Sampling volumes are programmable from 1 – 1,000 litres with 5 preset volumes to ensure reproducible results. The unit is pre-calibrated at Merck Millipore but re-calibration on-site is easy withthe addition of our DA-100® Digital Anemometer.13The MAS -100 CG Ex®The MAS-100 CG Ex ®Compressed Gas Microbial Air SamplerThe MAS-100 CG Ex® System samples at two constant flow rates, 100 L/min and 50 L/min for low flowapplications. The system will sample gas at a pressure range between 1.5 bar to 10 bar. Compressed gas isdirected through a perforated plate onto the 90 – 100 mm Petri dish. After sampling of the required volume of gas, a gradual decompression occurs automatically, preventing any sudden pressure change, avoiding possible damage to the microorganisms and improving recovery. The MAS-100 CG Ex® system is the onlycompressed gas microbial Air Sampler approved for use in Zone 2 explosion hazard areas.The unit is pre-programmed for compressed air, nitrogen, carbon dioxide and argon gas, and a total of 10 gas protocols can be programmed and stored in the unit.The automated collection process saves time and eliminates the awkward and risky manipulations required with manual methods.Technical specifications for MAS-100 CG Ex®Coated aluminum100 litres/min. + 10 % (over the pressure range) 1.5 bar to 10 bar (absolute)50 litres/min. + 10 % over the pressure range of 1.5 bar to10 bar (absolute) 50, 100, 250, 500, 1,000 litres1 to 2,000 litres, volumes individually selectable between 0 and 2,000 litres. 0 volumes are not displayed Technical specifications for Sampling Head14T h e D A -100N T ®The DA-100 NT ®Automatic calibration featureThe flow rate accuracy is ± 1.0 %, the most accurate on the market. The display also shows temperature (°C), velocity in m/sec and barometric pressure in mbar. The DA-100 NT® may be used in manual or automatic calibration modes for calibration of the MAS-100 NT® instruments.The DA-100 NT® is calibrated at Merck Millipore laboratory using a critical venturi flow standard, SWISS traceable. Certification is provided for each instrument.The DA-100 NT® Digital Anemometer is a highly accurate transfer standard used to calibrate both MAS-100® and MAS-100 NT® instruments (cannot be used on the MAS-100 CG Ex®). This digital vane anemometer spins freely on a magnetic field bearing and the revolutions are counted by a laser diode. Technical specifications for DA-100 NT®* NIST=National Institute of Standards and Technology15Services & SupportServices plans: Preventative & Curative MaintenanceMerck Millipore has the most comprehensive preventative and curative maintenance program available for microbial air sampling.The services provided include:• C omplete yearly visual, functional and perfor-mance “as found” and “as left” checks including calibration for all MAS-100® systems. • C alibration certificate (traceability fromGFW Switzerland) and complete service report is provided with every services.In addition to the above services, additional verification, calibration or preventive maintenance (included verification & calibration) options are available. We offer several contract levels to give you the possibility to enhance the coverage of the equipment e.g. spare part and repair visit can be included in your contract.Our certified service engineers can service your air sampler in our closest repair depot or directly in your lab. Contact your local sales representative for more information.Validation & Training SupportMerck Millipore aims to provide complete validation documentation to help you through the validationprocess. Our validation protocols are based on our internal product qualification test methods. These extensive protocols will enable the QC/QA Lab to quickly initiate your validation master plan and perform IQ, OQ and PQ (suitability of the test methodology) with ease.They follow international guidelines such as EP/USP and GMP.To confirm the availability of validation support for your needs, please contact your local sales representative.Traceability & Calibration AccuracyThe sample collection volume for each MAS-100® system is measured and adjusted with the DA-100 NT® System (except the MAS-100 CG Ex®), which is directly traceable to standards from GFW Switzerland. Traceability is not a guaranty of measurement accuracy, it is a chain of documentation between the last measurements made, linking it to the referenced standard. The uncertainty (accuracy) of a measurement is determined by the combined uncertainties of all measurements made by devices between the referenced standard and that devise being calibrated, this is called the “chain of comparisons”. With each link in the chain, additive uncertainty occurs. To minimize uncertainty and maintain accuracy the number of links in the chain of comparison must be minimized and the uncertainty for each link must be documented.This is the highest level of flow accuracy available from any microbial air sampler manufacturer. You can have the confidence when monitoring critical environments that every sample volume collected is accurate.Merck Millipore has experienced and trained validation engineers who are skilled to assist in validation protocol implementation within the QC microbiology laboratory, so the QC/QA departments do not have to allocate resources. A complete and technical training on your Merck Millipore air sampler is also provided during the validation engineer’s visit. Having a Merck Millipore validation protocol and on-site service eliminates high costs, both apparent and hidden, and helps ensure the validation is completed quickly and economically and guarantee optimal performance over the equipment life time. Contact your local sales representative for the availability of training at your site.16ICR and ICRplus Settle PlatesThe ICR and ICRplus Settle Plates are produced under aseptic conditions, gamma-irradiated and triple-bagged, making them optimal for active air monitoring with MAS-100® Microbial Air Samplers. All of the MAS-100® Air Samplers have been validated according to ISO 14698 using ICR and ICRplus Settle Plates – they all showed the same reliable results in physical and biological efficiency testing.I C R a n d I C R p l u s S e t t l e P l a t e s17Close-up of the new two way closure systemFor use in isolators and cleanrooms • O ne product for all applications •V HP-impermeable packagingRecommended Casein Soya Bean Digest Agar (TSA) acc. to USP and FDA • F ormulations with neutralizers forthe inactivation of disinfectant • A dd-On: Chocolate Agar + LTH – ICRplusfor the detection of fastidious bacteriaRoom temperature storage• S torage at site of use • L ess cooling capacity neededICRplus Settle Plates – Secure your samples during transport with the innovative two way closure system • C LOSED-position for safe transportand aerobic incubation • V ENT-position for anaerobic/microaerophilic incubation18MAS-100 NT® Air Samplerwith hard carrying case, battery, 300-hole perforated lid, dust cover, mains charger, USB cable, 3 mm allen key, and operator’s manual on CD 1.09191.0001MAS-100 NT® Perforated Lid, Aluminum, 300-hole 1.09195.0001MAS-100 NT® Perforated Lid, Aluminum, 400-hole 1.09088.0001MAS-100 NT® Perforated Lid, Stainless Steel, 400-hole 1.09424.0001MAS-100 NT® Dust Cover, Aluminum 1.09084.0001MAS-100 NT® Li-Ion Battery 1.09208.0001MAS-100 NT® Mains Charger1.09200.0001MAS-100 NT® Ex Air Sampler, Explosion Proofwith hard carrying case, battery, 300-hole perforated lid, dust cover, mains charger, USB cable, 3 mm allen key, and operator’s manual on CD 1.09194.0001MAS-100 NT® Ex Perforated Lid, Aluminum, 300-hole 1.09195.0001MAS-100 NT® Ex Perforated Lid, Aluminum, 400-hole 1.09088.0001MAS-100 NT® Ex Dust Cover1.09084.0001MAS-100 Eco® Air Samplerwith hard carrying case, battery, 400-hole perforated lid, dust cover, mains charger, 3 mm allen key, and operator’s manual on CD 1.09227.0001MAS-100 Eco® Perforated Lid, Aluminum, 400-hole 1.09088.0001MAS-100 Eco® Dust Cover, Aluminum 1.09084.0001MAS-100 Eco® Mains Charger 1.09128.0001MAS-100 Eco® Tripod Adapter 1.09127.0001MAS-100 Eco® Dust Cover1.09084.0001MAS-100 CG Ex® Air Samplerwith sampling head (50 L/min), hose, mains charger, operator’s manual on CD 1.09327.0001MAS-100 CG Ex® Sampling Head1.09237.0001MAS-100 Iso NT MH® Control unit, 1 head1.17174.0001MAS-100 Iso NT MH® Control unit with Profibus, 1 head 1.17177.0001MAS-100 Iso NT MH® Control unit with Ethernet, 1 head 1.17178.0001MAS-100 Iso NT MH® Control unit, 2 heads1.17118.0001MAS-100 Iso NT MH® Control unit with Profibus, 2 heads 1.17144.0001MAS-100 Iso NT MH® Control unit with Ethernet, 2 heads 1.17145.0001MAS-100 Iso NT MH® Control unit, 3 heads1.17146.0001MAS-100 Iso NT MH® Control unit with Profibus, 3 heads 1.17147.0001MAS-100 Iso NT MH® Control unit with Ethernet, 3 heads 1.17148.0001MAS-100 Iso NT MH® Control unit, 4 heads1.17149.0001MAS-100 Iso NT MH® Control unit with Profibus, 4 heads 1.17155.0001MAS-100 Iso NT MH® Control unit with Ethernet, 4 heads 1.17157.0001MAS-100 Iso NT® Isolator System 1.09168.0001MAS-100 Iso NT® Control unit with Profilbus 1.09173.0001MAS-100 Iso NT® Control unit with Ethernet 1.09174.0001MAS-100 VF®1.17103.0001MAS-100 VF® power supply 1.17104.0001MAS-100 VF® Petri dish clamps, 3 1.17171.0001Quick Adaptor for Tripod1.09223.0001Ordering InformationAccessoriesMAS-100 Iso NT® IP54 Kit 1.17159.0001MAS-100 Iso NT® Perforated lid, stainless steel, 300-hole edge protected 1.09189.0001MAS-100 Iso NT® Perforated lid, stainless steel, 400-hole edge protected 1.09222.0001MAS-100 Iso NT® Base for sampling head, stainless with Tri-Clamp and gasket 1.09328.00011.17091.0001MAS-100 Iso NT® Easy clean base for sampling head, stainless steel,with 1 Tri-Clamp & 1 gasketMAS-100 Iso NT® Perforated lid, stainless steel, 300-hole 1.09329.0001MAS-100 Iso NT® Perforated lid, stainless steel, 400-hole 1.09424.0001MAS-100 Iso NT® Tri-Clamp, stainless steel 1.09440.0001MAS-100 Iso NT® Dust Cover, stainless steel 1.09644.0001MAS-100 Iso NT® Remote Control 1.17181.0001MAS-100 Iso NT® Elbow Joint with 2 Tri-Clamps 1.17083.0001MAS-100 Iso NT® Power Supply 1.17182.0001MAS-100 Iso MH® Power Supply 1.09784.0001MAS-100 Iso NT® Silicone Gasket 1.170.840.001MAS-100 Iso NT® Pressure Test Kit 1.170.850.001MAS-100 Iso NT® Silicone gaskets for easy clean base 1.170.990.0014" Opticap XL 4KTGRA04TT35" Opticap XL 5KTGRA05TT1M Air T® Isolator Tubing 3 m ATBTUBE01Flexable tube connector PF01501Tripod 1.09326.0001Tripod adaptor for MAS-100 Eco® 1.09127.0001MAS-100® Mains Charger 1.09085.0001MAS-100® Tube Adaptor 1.09224.0001Contact Plate Adaptor 1.09214.0001Perforated Lid for Contact Plates 1.09213.0001MAS-100® NiMH Battery Pack (for Version 2.7x or higher) 1.09229.0001MAS-100® Power supply 1.09085.0001Note: The 300-hole perforated lid is compatible with legacy systems, but a specific calibration is requiredValidation Protocols (A4)MAS-100 VF® Functional testing (A4)MAVFA4FT1MAS-100 Iso NT® Validation Protocol (A4)MAISA4VP1MAS-100 Iso MH® Validation Protocol (A4)MAMHA4VP1MAS-100 NT® Validation Protocol (A4)MANTA4VP119ReferencesAnderson, A.A. (1958)“New Sampler for the collection, sizing and enumeration of viable airborne particles.”Journal of Bacteriology, Vol 76, pp. 471-484Meier, R., Zingre, H. (2000)“Qualification of Air Sampler systems: The MAS-100®” Swiss Pharma 22, No. 1-2, pp. 15-21Feller, W.“An Introduction to Probability Theory and itsapplications”, John Wiley and Sons Inc., New York, 1950"Microbial Control and Monitoring of Aseptic Processing Environments" U.S. Pharmocopiea (USP), General Chapters <1116>, 2012 Ewald, R., and Meier, R. 2004“Detection of micro-organisms in compressed gases: Validation of MAS-100® CG using compressed gases.” Swiss Pharma, 26(10a): 16-18Merck KGaAFrankfurter Straße 25064293 Darmstadt, Germany/officesFor more information on our products:/biomonitoringFind contact information for your country at: /offices For Technical Service, please visit: /techserviceThe M logo is trademark of Merck KGaA, Darmstadt, Germany. MAS-100® is a registered trademark of MBV, Staefa, Switzerland. All other trademarks are the property of their respective owners. Lit No. PB4667EN00 08/2015© 2015 Merck KGaA, Darmstadt, Germany. All rights reserved.We provide information and advice to our customers on application technologies and regulatory matters to the best of our knowl-edge and ability, but without obligation or liability. Existing laws and regulations are to be observed in all cases by our customers. This also applies in respect to any rights of third parties. Our information and advice do not relieve our customers of their own responsibility for checking the suitability of our products for the envisaged purpose.。

流式细胞术最新进展及临床应用

流式细胞术最新进展及临床应用流式细胞术( F l o w cy t o m e t r y, F C M), 临床上也被称为流式细胞分析,是利用流式细胞仪同时对单个细胞的多个参数进行定性/ 定量( 相对/ 绝对) 分析的生物医学分析技术,检测速度快、通量高、灵敏度高、采集数据量大、节约样本及成本,在临床上已经广泛应用于血液学、免疫学、肿瘤学、精子学等检验领域,是未来临床检验不可替代的检测方法之一。

传统流式细胞术,也被称为荧光流式细胞术,是基于荧光标记及荧光发射光谱检测的一门综合性技术,定量方式多为定性分析,检测参数类型单一、数目有限,数据分析复杂且缺乏标准化分析流程,不同检测中心间数据重现性差,这些都限制了它在临床检验中进一步的推广及应用。

近年来,为克服以上问题,流式细胞术不断突破与创新,从定性检测发展为定量检测;从单参数分析、双参数分析发展成为多参数分析;从检测细胞表面抗原到胞内抗原及分泌到胞外的抗原;从检测蛋白表达水平发展为检测蛋白定位、蛋白功能及蛋白翻译后修饰等;从一维定量检测发展为二维定量定位分析,从体外检测发展为体内检测等;这些突破使得流式细胞术可以实现从单细胞水平去认识细胞在生理或病理状态下的免疫表型、分子表型甚至各种复杂的信号通路变化等,因此将更为广泛应用于临床检测。

1定量流式细胞术定量流式细胞术( Q u a n tit a ti ve fl o w cy t o m e t r y, QFCM),即通过流式细胞仪定量检测细胞或微球上荧光素的中值荧光强度 ( M e d i a n fl u o r e s ce n t i n t e n s it y, M F I ) 或每个细胞结合的抗体单位( A n ti b o d i e s b o und t o p e r ce ll,A BC) 来对生物分子进行相对或绝对定量的流式细胞技术。

定量流式细胞术已被证明是一种功能强大的临床检验技术,但由于M F I缺乏标准化度量方法,容易引起不同检测中心检测结果重现性差,导致诊断和治疗决策的不确定性及不可靠性, 限制了其在临床的推广应用, 因此,标准化M F I测量为流式细胞术实现精确定量分析,在临床广泛应用的必经之路。