guava easyCyte CN 20110419 - antpedia.com
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guava® easyCyte微毛细管细胞分析平台
Guava 细胞分析平台
作为生命科学和治疗领域的提供者和合作
者,默克密理博公司致力于细胞生物学与免疫
学研究领域解决方案的研究与开发。Guava系
列细胞分析仪是为细胞生物学与免疫学实验
研究专门设计的。Guava仪器紧凑集约型的设
计,易于掌握的操作功能,强大分析能力,实
现了集仪器、软件和试剂于一体的完整细胞分
析研究平台。
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Guava easyCyte 系列微毛细管细胞分析
平台采用目前先进的荧光检测系统和优质的光
学组件以期最大可能的提高检测灵敏度。程序
化软件模块避免操作错误,将给出精确的和可
重复的数据。专业设计,大大简化了复杂的操
作流程,轻松完成开机、样品检测和关机维
护。
easyCyte 8HT双激光六色八参数高通量
easyCyte 8双激光六色八参数高通量
easyCyte 5HT单激光三色五参数高通量
easyCyte 5单激光三色五参数高通量
easyCyte 6HT单激光四色六参数高通量
easyCyte 6单激光四色六参数高通量
easyCyte 6HT-2L双激光四色六参数高通量
easyCyte 6-2L双激光四色六参数高通量
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独创的微毛细管上样
每一台Guava系统均采用了全世界独有微毛细
管液流 (上样) 设计,摒弃了传统的鞘液流方
式,系统不需要使用鞘液即可形成稳定的单细
胞流。由于Guava采用微毛细管设计,使最小
上样量可达到1000个细胞/Test。微毛细管上
样设计使得上样细胞无损失,同时高精确的注
射泵上样,体积精确可控。
微毛细管核心技术带来的优点:
• 固定化细胞流路,液流自动校准,操作简捷;
• 样品需求少 (最小上样量1000个细胞/Test),
节省试剂和样本;
• 无需使用鞘液,最大程度减少废液的产生
(每运行8小时的废液产生量不超过50ml),减
小了仪器的体积;
• 精度注射泵与PEEK管路系统双保险,较少
Clog的烦恼;
• 上样细胞无损失,上样体积精确可测,实现
绝对细胞计数;
• 直接从样品管中吸取细胞样本。
与传统基于鞘液流体动力学的细胞上样模式,
无鞘液的微毛细管上样系统。样品上样时,由
微毛细管与样品液体接触的横截面中心形成的
负压区,形成单细胞流上行。注射泵提供稳定
精确的上样流与上样体积。由微毛细管和注射
泵协同作用形成的单细胞流更加精确与稳定。
每运行8小时的废液产生量不超过50ml。上样
体系小,每个test最小上样量仅为1000个细
胞,这使您可以减少样本的需求量、节约样本
处理时间与抗体试剂的消耗,从而提高您的研
究效率以及节省您的实验室运营成本。
Sheath Fluid: ~10 mL/testWaste: 8,000 mL per 8 hour runTypical #Cells Per Protocol: 100,000 - 1,000,000 cells/test
Traditional
Waste
SampleSheath Fluid
Laser
Laser
Shea
th F
low
Inje
ctio
n T
ip
Waste
Sam
ple
Flow
Sheath Fluid: NoneWaste: < 50 mL per 8 hour runTypical #Cells Per Protocol:≥1,000 cells/test
Microcapillary
Sample
Laser
WasteLaser
Waste
Flow Cell
Sample
Sam
ple
Flow
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Guava 细胞分析检测系统
专利的微毛细管技术和激光系统保证了仪器能
够检测包括哺乳动物细胞、微生物、藻类、细
菌以及微珠在内的多种样本。在检测过程中,
荧光标记的细胞首先被吸入微毛细管上样针
内,通过流动室时,蓝色激光或红色激光并线
激发细胞上的荧光素,其产生的散射光信号被
光电倍增管等电子器件捕捉。经过数字处理
后,通过GuavaSoft软件模块,将数字信号呈
现在计算机上,以进行数据分析和处理。
液流系统
Guava的液流系统由微毛细管、注射泵、废液
瓶、管路组成。由微毛细管与样品液体接触的
横截面中心形成的负压区,使单细胞流更加精
确与稳定。注射泵提供稳定精确的上样流与上
样体积,保证了Guava可以在不需要使用任何
参考微珠的情况下进行绝对细胞计数。由于
系统不需要使用鞘液,每运行8小时所产生的
废液仅有50ml,极大地减少了废液处理带来
的工作量。系统内部管路材料均使用PEEK材
料,PEEK对蛋白与颗粒均不吸附,管路清洗
异常简单方便。
光路设计
全固态激光器
Guava系列微毛细管细胞分析系统均采用全固
态激光器。全固态激光器 (DPL) 具有体积小、
重量轻、效率高、性能稳定、可靠性好、寿命
长、光束质量高等优点。半导体激光泵浦的全
固态激光器是20世纪80年代末期出现的新型激
光器。全固态激光器由于其单位输出的热负荷
降低,可获取更高的功率,更长的系统寿命和
可靠性。使用寿命超过25000小时。
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sample
flow cell
waste
Laser
独特的双激光共线性设计
Guava双激光系统 (easyCyte 6-2L/6HT-2L、
easyCyte 8/8HT) 采用了独特的双激光同步共
线性设计,两个波长的激光即不相互干扰也不
需要进行激光间时间延迟的校正。消除了以往
多激光光路设计中普遍存在的信号延迟误差问
题。
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Rainbow Bead Data
Rainbow Beads校正——流式细胞分析仪检测的金标准
Rainbow Calibration Particles from Spherotech - 8 Peak detection.SPHEROTM 多色荧光校验微珠是检验流式细胞性能的金标准。通过该校验能够测试仪器的灵敏度和线性信号。从图中独立的荧光波峰显示了仪器的高灵敏度,不存在信号间的交叉;FITC,PE,PE-Cy5.5清晰的八峰证明了仪器信号良好的线性度。
CD4 staining of Whole Blood across all 6 Fluorescent Channels
不同的荧光通道,相同的实验结果——完美配合您的实验设计
6种荧光染料检测全血种的T细胞实验过程:取成人全血,每10ul为一份,分别与CD4-FITC, CD4-PE, CD4-PECy5.5, CD4-PECy7, CD4-APC, CD4-APCCy7混合,室温孵育15分钟。每份样品中加入180ul,含0.2% PFA 固定液的细胞裂解液。 在easyCyte 8HT中上样检测CD4%和CD4T细胞记数。如图表所示,可在所有的6个荧光通道中都可以分到清晰的CD4群体,得到细胞绝对记数,和CD4百分比结果。
精密设计的数字化光电转换系统
Guava精密的数字化光化电转换系统数据获取
速度更快,拥有数字化的光电转换系统,荧光
间的补偿调节更加简单,激光内的荧光补偿可
自由进行,可实现实时在线补偿与离线补偿。
进样系统——高通量自动进样
• 上样系统预设了1块96孔板与10个单样品管
(1.5ml或0.5ml tube) 的位置,使得 高通量
上样与中低 通量上样可同时完成,避免了从
中低通量到高通量上样时需要手动切换所带
来的麻烦。
• 特有的桨式mixer为每个样品提供上样前的
再重悬,避免样品在等待过程中产生下沉。
保证所有样品都是在均匀悬浮状态时上样,
从而减小孔间误差。
• 自动上样程序整合了样品间 清洗程序,同时对
上样针与mixer进行清洗和空干,极大程度地
避免了样品间交叉污染 (carryover<0.05%)。
进样系统——单管上样系统
• 上样系统预设了2个单样品管 (1.5ml或0.5ml
tube) 的上样位置,手动可以进行快速上样
与样品读取。
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InputSignal
Amplifier FastADC
DSP*
* digital signal processor
height
width
area
Com
pute
r
Wash tubes
Plate holder
Tubes positions
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GuavaSoft 数据采集和分析系统
GuavaSoft 是一套专为细胞生物学研究量身
定制的软件。包括了无需补偿设置的模块化软
件GuavaSuite,拥有强大图形显示和数据分
析能力的InCyte,以及广受好评的ExpressPro
操作软件。集数据获取、数据分析和仪器维护
程序于一体 。通过GuavaSoft 友好的视窗操
作界面, 使数据获取和实验分析的操作更为
灵活。通过对多种复杂、交叉的数据进行更深
入、更立体的分析,协助您完整地了解细胞生
物学事件。
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GuavaSuite Assay-specific Software Modules ——优化过的自动分析模块
GuavaSuite由一系列特异性模块组成,为凋
亡研究、细胞活力分析和细胞周期等应用分别
专门设计的实验方案、补偿参数及分析模板。
由于每个模块只针对一种特异性应用,其独特
的补偿优化设计,使您无需进行复杂的补偿调
节*;预设的实验方案,自动设置标准化结果
呈现形式;分析模块内嵌,自动计算数据结
果。所有这些只为加快您的实验过程,得到更
精确的数据。
*需配合Guava的试剂一起使用。
GuavaSuite 特点:
• 针对性强,一个模块专为一类研究而设
• 预设实验方案,补偿内置,一键式操作完成
数据获取
• 自动呈现标准化数据窗口,无需手动设置数
据的横纵坐标
• 分析模板内嵌,针对不同研究分别优化的分
析功能,使结果更准确
• 批处理功能,96个数据分析一键完成
• 自动保存为Excel格式,可以FCS2.0格式输
出
GuavaSuite功能化模块包括:
• 细胞绝对计数和活力分析 (ViaCount)
• 早期凋亡Annexin V
• 线粒体膜电位
• 活化的Multi-Caspase检测
• 晚期凋亡 Tunel
• 细胞周期 Cell Cycle
• 细胞毒实验
• 细胞示踪
• 细胞增殖
• 抗体定量检测
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Guava InCyte 软件:流式细胞分析的新高度
——多维分析,解析高通量数据信息
InCyte是第一款集合了多种统计方法于流式分
析的软件。它为您提供特异性的图示分析功
能,整合6×96个生物信息,在Heat Map上直
观地显示数据间的关联性,协助您快速判断分
析结果。引入拖曳式Gating模式,自动嵌入内
在逻辑关联;自动批处理功能,96个数据分
析一次完成。InCyte所提供的数据获取和分析
功能,已远远超出了对一个样品进行统计的层
面,已然成为一款理想的,从生物信息学角度
解析多参数、多样本间关联性研究的平台,适
用于siRNA筛选,药物研发中化合物的筛选以
及其他高通量的细胞生物学检测。
Heat Map (热图):
Heat: 由浅至深的蓝色代表了信号的强/弱。颜
色愈深,则信号愈强。
Map: 显示样品在96孔板中的位置,直观显示
样品名。同时,根据所检测的指标 (1-6
个),将样品孔相应地分割成1-6等分的
饼图。
通过HeatMap的诠释,可比较96孔板上不同
样品的检测结果,而且可以整合1至6张96孔
板上的信息,即比较6×96 个样品的数据,
显示多参数信号的强/弱,展现各参数间的关
联。直观的图示能够有效地协助您做出快速、
准确的判断。
以药物研发中化合物的筛选为例:
Screen compounds for Apoptosis induction
in Jurkat Cells. Want to find compound
that induces apoptosis via multiple markers
in Jurkat cells. Apoptosis Markers Tested:
Caspase 8, Caspase 9 , Mitochondrial
depolarization and Annexin V
Set Up:Four 96 well plates:
Plate 1 - Jurkat Cells + compounds –
Caspase 8 Reagents
Plate 2 - Jurkat Cells + compounds –
Caspase 9 Reagents
Plate 3 - Jurkat Cells + compounds – Mito
Potential Reagents
Plate 4 - Jurkat Cells + compounds – Nexin
Reagents
The Heat-Map displays comparative findings
in Jurkat cells for all 4 apoptotic markers
(Legend inset). Certain compounds
demonstrated Caspase-8 (F9, H11) or
Caspase-9-(B11, C4) specific modes of
induction while others invoked cell-specific
caspase pathways. Discrimination required
early assessment (4HR) as both enzymes
are actively involved during later stages of
cell death. At 10uM, JC-1 staining, provided
no additional information.
Drag-and-drop gating from one plot to another
View up to 11 plots at once
Real-time plot adjustmentsOrganize
acquired data sets and select individual wells for display
Quickly link to and review previously analyzed data
Easily create analysis templates
Combine groups of data and analysis templates to construct heat maps or EC50 curves
Slider bars set cut-offs or threshold values for each experimental sector
Heat map shows values across an entire plate
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Drag and Drop Gating (拖曳式设门) ——操作简单,逻辑关系明确。
IC50/EC50自动计算
独创将IC50/EC50分析功能直接嵌入流式分析
软件。可使用细胞增值或凋亡数据直接计算
IC50/EC50,使您的实验更加快捷、高效。并
且无需您额外购买第三方分析软件。
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友好的互动式视窗操作界面
• 可实时监控11个数据窗口
• 阶梯式数据管理模式,可整合六张96孔板数
据,以实现对6×96个数据的整合分析。
• 可实现在线补偿和离线补偿
• 数据可直接以FCS文件形式导出,与第三方
分析软件兼容。数据可自动保存为Excel格
式,图片输出与Microsoft office软件兼容。
ExpressPro 8.0 ——全开放平台,灵活配合您前沿的研究需求
ExpressPro是一种全开放数据获取和分析软
件,提供补偿调节,电压调节,脉冲调节;
Lin/Log数据格式选择;可自由设计实验方案
和分析模板等功能,以配合您前沿的研究需
求。
• 可同时监控8个窗口
• 五种Gating方式可选
• 批处理功能,多个样品一键分析
• 可通过overlay进行样品间的比较
• 数据可输出为FCS2.0、Excel格式
• 与自动化平台兼容:Robotic arms;Liquid
Handling Stations
功能
通过overlay进
数据可输出为FCS2.0
与自动化平台兼容:Robo
Handling Stations
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系统使用与维护
由于无需鞘液,Guava 系列细胞分析仪无需对
复杂的鞘液系统进行维护。固化的细胞流路实
现了细胞流自校准。PEEK管路材料和高压注
射泵保证了液流系统的顺畅。自动Easy Check
功能是简单程序化的QC工具,使您可以在任
何时间对仪器工作性能进行检测。
无需鞘液,废液产生少
无需专人维护,免去繁复的鞘液检查与机械
校正;系统每运行8个小时废液排放量小于
50mL,更能免去因废液处理而给您带来的额
外的工作。
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Easy Check 实现程序化QC过程
Easy Check试剂盒使用标准化的荧光Beads
与试剂通过精确计算和荧光检测来验证系统工
作是否正常。使用Easy Check可以将您从繁
复的校检工作中解脱出来,实现机器自动自检
功能。
一键式关机操作
—— Clean & Shut down使用Cleaning功能可以自动完成仪器的清洁,
确保仪器的正常使用。在一天实验结束后运行
Cleaning能够有效地避免荧光染料在流动室中
的残留,并能有效的清洁整个管路系统,从而
保证了仪器的正常使用。
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默克密理博为众多细胞分析应用提供一系列全
套的Guava微毛细管细胞分析系统与服务,这
些产品秉承了默克密理博对用户的一贯:即提
供质量优秀、开放灵活、使用简便和低成本的
产品。默克密理博所提供的微毛细管细胞分析
系统包括:Guava微毛细管分析系统主机、专
业分析软件、专业技术服务与相关应用的专业
试剂。
Guava easyCyte HT 系列为客户提供了一个自动上样托盘,可以同时自动处理96孔微孔板
和多达10个样品管的高通量分析。
Guava easyCyte Mini 系列为客户提供了一个可进行手动上样的上样托盘,可以处理
2个样品管。
Microcapillary fl ow cellrequires no sheath fl uid and is
user-replaceableSingle sample loaderSwivel arm functionality, holds two tubes and allows instant acquisition
Waste vial collects less than 80 mL of waste in a typical 8-hour workday
Wash vial offers a high-pressure purge to easily clear obstructions from the fl ow cell
Up to six-color detectionmade possible by one (blue) or
two excitation lasers (blue and red)
Small footprint saves valuable laboratory spaceWidth: 17.75 in (45.1 cm)Depth: 17.25 in (44.5 cm)Height: 8.75 in (22.2 cm)(does not include laptop)
Microcapillary fl ow cellrequires no sheath fl uid and
is user-replaceable
Wash vial offers ahigh-pressure purge toeasily clear obstructionsfrom the fl ow cell
Waste vial collects lessthan 80 mL of waste in atypical 8-hour workday
Robotic sample trayprovides walk-awayautomation for a 96-wellmicroplate and up to 10sample tubes
Up to six-color detectionmade possible by one (blue) or
two excitation lasers (blue & red)
Small footprint savesvaluable laboratory space
Width: 20.3 in (51.5 cm)Depth: 23.4 in (59 cm)
Height: 10.0 in (25.4 cm)(does not include laptop)
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Guava easyCyte 8HT 和 NEW easyCyte 8微毛细管细胞分析系统
Guava easyCyte 8HT与easyCyte 8均配置了
488nm蓝色固态激光器与640nm红色固态激
光器,可同时提供多达8个检测参数,包括6个
荧光参数与前向散射角和侧向散射角2个光学
参数。
可检测的荧光素:FITC, PE, PE-Texas Red®,
PerCP or PerCP-Cy5.5, PE-Cy7, APC, Alexa
Fluor® 700, APC-Cy7。
583 nm
Side Scatter
Forward ScatterBLUE LASER (488 nm)
Multiwell Plate
RED LASER (640 nm)
525 nm
690 nm
661 nm785 nm
785 nm
RED1
NIR – Near InfraRed FITC – FluoresceinPE – Phycoerythrin APC – Allophycocyanin
YELLOW NIR1
BLUE LASER (488 nm)
GREEN
FITC PE PE-Cy5PE-Cy5.5
PE-Cy7
RED2 NIR2
RED LASER (640 nm) APC APC-Cy7
Inside the guava easyCyte 8HT and easyCyte 8 systems
HOW IT WORKSThe guava easyCyte systems use patented, microcapillary, laser-based technology capable of detecting mammalian and microbial cells and beads. A sample of fl uorescently labeled cells is aspirated into a uniquely proportioned microcapillary fl ow cell. A red or blue diode laser excites the cells and each cell emits signals that are individually detected by photomultipliers and a photo diode. Guava software modules show all relevant data and results immediately.
SPECIFICATIONS
System Guava easyCyte 8 Guava easyCyte 8HT
Catalogue # 0500-5008B 0500-4008B
LaserBlue Laser solid state 488nm 75mWRed Laser solid state 640nm 40mW
Blue Laser solid state 488nm 75mWRed Laser solid state 640nm 40mW
Laser ConfigurationSpatially separated beams with 20 x 700-μm elliptical spots
Spatially separated beams with 20 x 700-μm elliptical spots
Forward Scatter Detector √ √
Side Scatter Detector √ √
Fluo
resc
ence
Det
ecto
rs
PM
T
GREEN 525/30 nm 525/30 nm
YELLOW 583/26 nm 583/26 nm
RED 1 690/50 nm 690/50 nm
NEAR INFRARED 1
785/70 nm 785/70 nm
RED 2 661/19 nm 661/19 nm
NEAR INFRARED 2
785/70 nm 785/70 nm
Sample Flow Rate 7 μL/min to 71 μL/min 7 μL/min to 71 μL/min
Sample Acquisition Rate10,000 events/second, 6 compensated fluorescence parameters and 2 scatter parameters
10,000 events/second, 6 compensated fluorescence parameters and 2 scatter parameters
Sample Requirement ≥1000 cells/test ≥1000 cells/test
Particle Size 0.2μm - 70μm 0.2μm - 70μm
Automation – 96 well and 10 tubes
N/A √
Mixing N/A √
Carryover N/A < 0.05%
Pulse Processing Digital Signal Processing Digital Signal Processing
Fluorescence Threshold Sensitivities
FITC <100 MESF FITC <100 MESF
System SizeHeight: 10 in (25.4 cm)Width: 20.3 in (51.5 cm)Depth:23.4 in (59 cm)
System Weight 73 lbs (32.4 kg)
18
19
100 101 102 103 104
Side
Sca
tter-(
SSC-
HLin
)
Plot P01: Ungated
CD45 PerCP-Cy5.5 (RED-HLog)
050
0040
0030
0020
0010
0060
0080
0070
0090
00
CD45+
100 101 102 103 104
050
0040
0030
0020
0010
0060
0080
0070
0090
00
Side
Sca
tter-(
SSC-
HLin
)Plot P02: Gated on CD45+ cells
CD3 FITC (GRN-HLog)CD3+
CD3-
104100 101 102 103
CD19
APC
(RD2
-HLo
g)
Plot P04: Gated on CD3- cells
CD16+56 PE (YLW-HLog)
100
101
102
103
104
B-Cells
CD4
PE-C
y7 (N
IR-H
Log)
Plot P03: Gated on CD3+ cells
CD8 APC-Cy7 (NI2-HLog)100 101 102 103 104
100
101
102
103
104
CD3+ & CD4+
CD3+ & CD8+
NK Cells
Immunological Phenotyping-6 Color Assay
50 μL adult human blood was stained for 15 minutes at room temperature with a cocktail containing CD3-FITC, CD16-PE, CD56-PE, CD45-PerCP-Cy5.5, CD4-PE-Cy7, CD19-APC, and CD8-APC-Cy7. After incubation, cells were lysed and fi xed with 900 μL guava lysing solution containing 0.2% PFA. Samples were then acquired on the guava easyCyte 8HT system. CD45 positive cells were gated into a SSC vs. CD3 (FITC) plot. Lymphocytes (CD3+ and CD45+) were gated into a plot comparing CD4 and CD8 positive cells. To separate the natural killer (NK) and B cells from the lymphocytes, CD3-negative cells were gated into a plot comparing CD19 (B-cells) and CD16+56 (NK cells). The counts and percentages for each application were calculated. As the fi gure shows, separation is visible and is comparable to other published data.
Parameter Percent (%) Absolute CountLymph Events 2862
CD3+ 76.76 2197
CD3+ CD8+ 32.41 712
CD3+ CD4+ 34.55 759
CD3+ CD4+ CD8+ 1.82 40
CD16+ CD56+ 7.3 210
CD19+ 12.5 357
4/8 Ratio 1.07
easyCyte 8
Guava easyCyte 8, Base System 0500-5008B
Guava® Flow Cell 0500-2270
Power Conditioner, International 1000-3340
GuavaSoft Software Package for Guava easyCyte Systems includes InCyte, Express Pro and GuavaSuite Modules
0500-5115
Guava® Instrument Cleaning Fluid (ICF) (100 mL)
4200-0140
Guava® Easy Check Kit 4500-0025
easyCyte 8HT
Guava easyCyte 8HT, Base System 0500-4008B
Guava® Flow Cell 0500-2260
Power Conditioner, International 1000-3340
GuavaSoft Software Package for Guava easyCyte Systems includes InCyte, Express Pro and GuavaSuite Modules
0500-4115
Guava® Instrument Cleaning Fluid (ICF) (100 mL)
4200-0140
Guava® Easy Check Kit 4500-0025
20
Guava easyCyte 6HT-2L 和 NEW easyCyte 6-2L微毛细管细胞分析系统
Guava easyCyte 6HT-2L与easyCyte 6-2L均配
置了488nm蓝色固态激光器与640nm红色固
态激光器,可同时提供多达6个检测参数,包
括4个荧光参数与前向散射角和侧向散射角2个
光学参数。
可检测的荧光素:FITC, PE, PE-Texas Red®,
PerCP or PerCP-Cy5.5 , APC, Alexa Fluor®
700。
583 nm
Side Scatter
Forward ScatterBLUE LASER (488 nm)
Multiwell Plate
RED LASER (640 nm)
525 nm
690 nm
661 nm
RED1
NIR – Near InfraRed FITC – FluoresceinPE – Phycoerythrin APC – Allophycocyanin
YELLOW
BLUE LASER (488 nm)
GREEN
FITC PE PE-Cy5PE-Cy5.5
RED2
RED LASER (640 nm) APC
Inside the guava easyCyte 6HT-2L and easyCyte 6-2L systems
HOW IT WORKSThe guava easyCyte systems use patented, microcapillary, laser-based technology capable of detecting mammalian and microbial cells and beads. A sample of fl uorescently labeled cells is aspirated into a uniquely proportioned microcapillary fl ow cell. A red or blue diode laser excites the cells and each cell emits signals that are individually detected by photomultipliers and a photo diode. Guava software modules show all relevant data and results immediately.
21
SPECIFICATIONS
System Guava easyCyte 6-2L Guava easyCyte 6HT-2L
Catalogue # 0500-5007B 0500-4007B
LaserBlue Laser solid state 488nm 20mWRed Laser solid state 640nm 40mW
Blue Laser solid state 488nm 20mWRed Laser solid state 640nm 40mW
Laser ConfigurationSpatially separated beams with 20 x 700-μm elliptical spots
Spatially separated beams with 20 x 700-μm elliptical spots
Forward Scatter Detector √ √
Side Scatter Detector √ √
Fluo
resc
ence
D
etec
tors
P
MT
GREEN 525/30 nm 525/30 nm
YELLOW 583/26 nm 583/26 nm
RED 1 690/50 nm 690/50 nm
RED 2 661/19 nm 661/19 nm
Sample Flow Rate 7 μL/min to 71 μL/min 7 μL/min to 71 μL/min
Sample Acquisition Rate10,000 events/second, 4 compensated fluorescence parameters and 2 scatter parameters
10,000 events/second, 4 compensated fluorescence parameters and 2 scatter parameters
Sample Requirement ≥1000 cells/test ≥1000 cells/test
Particle Size 0.2μm - 70μm 0.2μm - 70μm
Automation – 96 well and 10 tubes
N/A √
Mixing N/A √
Carryover N/A < 0.05%
Pulse Processing Digital Signal Processing Digital Signal Processing
Fluorescence Threshold Sensitivities
FITC <100 MESF FITC <100 MESF
System SizeHeight: 10 in (25.4 cm)Width: 20.3 in (51.5 cm)Depth:23.4 in (59 cm)
System Weight 73 lbs (32.4 kg)
22
小鼠CD4+ T细胞分别被定向诱导,分化为Th1, Th2, Th17细胞。经Fixable Viability Dye (FVD) 染料染色,区分活细胞与死细胞。
FSC vs SSC散点图用以界定淋巴细胞群。通过CD4-PerCP, IFN – γ PE, IL – 4 PE, IL – 17 PE 抗体分别鉴定Th1, Th2, Th17细胞。
easyCyte 6-2L
Guava easyCyte 6-2L, Base System 0500-5007B
Guava® Flow Cell 0500-2270
Power Conditioner, International 1000-3340
GuavaSoft Software Package for Guava easyCyte Systems includes InCyte, Express Pro and GuavaSuite Modules
0500-5115
Guava® Instrument Cleaning Fluid (ICF) (100 mL)
4200-0140
Guava® Easy Check Kit 4500-0025
easyCyte 6HT-2L
Guava easyCyte 6HT2L, Base System 0500-4007B
Guava® Flow Cell 0500-2260
Power Conditioner, International 1000-3340
GuavaSoft Software Package for Guava easyCyte Systems includes InCyte, Express Pro and GuavaSuite Modules
0500-4115
Guava® Instrument Cleaning Fluid (ICF) (100 mL)
4200-0140
Guava® Easy Check Kit 4500-0025
23
Guava easyCyte 5HT、easyCyte 6HT和 NEW easyCyte 5、easyCyte 6微毛细管细胞分析系统
Guava easyCyte 5HT、easyCyte 6HT、
easyCyte 5和easyCyte 6均配置了488nm蓝
色固态激光器,可同时提供多达5-6个检测参
数,包括3-4个荧光参数与前向散射角和侧向
散射角2个光学参数。
可检测的荧光素:FITC, PE, PE-Texas Red®,
PerCP or PerCP-Cy5.5, Alexa Fluor® 700,
Alexa Fluor® 750。
583 nm
Side Scatter
Forward ScatterBLUE LASER (488 nm)
Multiwell Plate
525 nm
690 nm
785 nm
RED1
NIR – Near InfraRed FITC – FluoresceinPE – Phycoerythrin
YELLOW NIR1
BLUE LASER (488 nm)
GREEN
FITC PE PE-Cy5PE-Cy5.5
PE-Cy7
Inside the guava easyCyte 8HT and easyCyte 8 systems
HOW IT WORKSThe guava easyCyte systems use patented, microcapillary, laser-based technology capable of detecting mammalian and microbial cells and beads. A sample of fl uorescently labeled cells is aspirated into a uniquely proportioned microcapillary fl ow cell. A red or blue diode laser excites the cells and each cell emits signals that are individually detected by photomultipliers and a photo diode. Guava software modules show all relevant data and results immediately.
24
SPECIFICATIONS
System Guava easyCyte 5Guava easyCyte 5HT
Guava easyCyte 6Guava easyCyte 6HT
Catalogue # 0500-5005B 0500-4005B0500-5005B/0500-5006B
0500-4005B/0500-4006
LaserBlue Laser solid state 488nm 20mW
Blue Laser solid state 488nm 20mW
Blue Laser solid state 488nm 20mW
Blue Laser solid state 488nm 20mW
Laser Configuration
Spatially separated beams with 20 x 700-μm elliptical spots
Spatially separated beams with 20 x 700-μm elliptical spots
Spatially separated beams with 20 x 700-μm elliptical spots
Spatially separated beams with 20 x 700-μm elliptical spots
Forward Scatter Detector √ √
Side Scatter Detector √ √
Fluo
resc
ence
D
etec
tors
PM
T GREEN 525/30 nm 525/30 nm 525/30 nm 525/30 nm
YELLOW 583/26 nm 583/26 nm 583/26 nm 583/26 nm
RED 680/30 nm 680/30 nm 680/30 nm 680/30 nm
NEAR INFRARED
N/A N/A 785/70 nm 785/70 nm
Sample Flow Rate7 μL/min to 71 μL/min
7 μL/min to 71 μL/min
7 μL/min to 71 μL/min
7 μL/min to 71 μL/min
Sample Acquisition Rate
10,000 events/second, 3 compensated fluorescence parameters and 2 scatter parameters
10,000 events/second, 3 compensated fluorescence parameters and 2 scatter parameters
10,000 events/second, 4 compensated fluorescence parameters and 2 scatter parameters
10,000 events/second, 4 compensated fluorescence parameters and 2 scatter parameters
Sample Requirement ≥1000 cells/test ≥1000 cells/test ≥1000 cells/test ≥1000 cells/test
Particle Size 0.2μm - 70μm 0.2μm - 70μm 0.2μm - 70μm 0.2μm - 70μm
Automation – 96 well and 10 tubes
N/A √ N/A √
Mixing N/A √ N/A √
Carryover N/A < 0.05% N/A < 0.05%
Pulse ProcessingDigital Signal Processing
Digital Signal Processing
Digital Signal Processing
Digital Signal Processing
Fluorescence Threshold Sensitivities
FITC <100 MESF FITC <100 MESF FITC <100 MESF FITC <100 MESF
System Size
Height: 10 in (25.4 cm)Width: 20.3 in (51.5 cm)Depth:23.4 in (59 cm)
Height: 10 in (25.4 cm)Width: 20.3 in (51.5 cm)Depth:23.4 in (59 cm)
System Weight 73 lbs (32.4 kg) 73 lbs (32.4 kg)
25
Jurkat细胞细胞凋亡检测。图A:Jurkat细胞未受凋亡诱导,正常组;图B:0.02 μm CD95/FASL刺激诱导凋亡; 图C:5 μM CD95/FASL刺激诱导凋亡。凋亡诱导后用Guava Nexin进行染色。从结果来看,当诱导剂剂量增加,凋亡细胞比例逐渐增加,而活细胞比
例由92.5%下降到45.1%
直方图示细胞色素c含量的变化。图A,Jurkat细胞用1 μM staurosporine诱导2小时后,细胞色素c含量与对照组的比较;图B,Hela细胞用3 μM staurosporine处理4小时后,诱导组与对照组比较;图C,HL-60细胞用33 μM anisomycin处理4小时后,诱导组与对照
组的比较; 图D, HB细胞用100 μM anisomycin处理4小时后,诱导组与对照组的比较。
26
TUNEL法检测凋亡晚期。图A,HL-60未经凋亡诱导;图B,HL-60细胞在经staurosporine诱导的结果。直方图中,BrdU+(绿色
门)为凋亡阳性,BrdU-(紫色门)为凋亡阴性。图C-F,HL-60经Camptothecin诱导4个小时,显示细胞在不同浓度诱导剂作用下,
凋亡细胞比例的变化
THP-1细胞经Guava Cell Cycle试剂盒染色30分钟后在检测。结果由Modifit软件的曲线拟合分析得各期的百分比
27
图A显示为流式结果图。BrdU-Alex Fluor488的使用能够更准确的标记S期。同时,通过PI染色区分G0G1期与G2/M期
easyCyte 5
Guava easyCyte 5, Base System 0500-5005B
Guava® Flow Cell 0500-2270
Power Conditioner, International 1000-3340
GuavaSoft Software Package for Guava easyCyte Systems includes InCyte, Express Pro and GuavaSuite Modules
0500-5115
Guava® Instrument Cleaning Fluid (ICF) (100 mL)
4200-0140
Guava® Easy Check Kit 4500-0025
easyCyte 5HT
Guava easyCyte 5HT, Base System 0500-4005B
Guava® Flow Cell 0500-2260
Power Conditioner, International 1000-3340
GuavaSoft Software Package for Guava easyCyte Systems includes InCyte, Express Pro and GuavaSuite Modules
0500-4115
Guava® Instrument Cleaning Fluid (ICF) (100 mL)
4200-0140
Guava® Easy Check Kit 4500-0025
easyCyte 6
Guava easyCyte 6, Base System0500-5005/0500-5006
Guava® Flow Cell 0500-2270
Power Conditioner, International 1000-3340
GuavaSoft Software Package for Guava easyCyte Systems includes InCyte, Express Pro and GuavaSuite Modules
0500-5115
Guava® Instrument Cleaning Fluid (ICF) (100 mL)
4200-0140
Guava® Easy Check Kit 4500-0025
easyCyte 6HT
Guava easyCyte 6HT, Base System0500-4005/0500-4006
Guava® Flow Cell 0500-2260
Power Conditioner, International 1000-3340
GuavaSoft Software Package for Guava easyCyte Systems includes InCyte, Express Pro and GuavaSuite Modules
0500-4115
Guava® Instrument Cleaning Fluid (ICF) (100 mL)
4200-0140
Guava® Easy Check Kit 4500-0025
www.millipore.com.cn客服电话: 400-889-1988
LS guava easycyte 02 2011年4月上海印刷
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