新世代流式細胞儀
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新世代流式細胞儀. Partec GmbH from Münster Germany. 派特科技有限公司. Fluorescence Based Flow Cytometry. Invented in 1968 in the University of M ün ster, Germany at the Institute of Radiobiology by Professor Dr.Wolfgang Göhde.

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新世代流式細胞儀

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Partec gmbh from m nster germany

新世代流式細胞儀

Partec GmbH

from Münster Germany

派特科技有限公司


Fluorescence based flow cytometry

Fluorescence Based Flow Cytometry

  • Invented in 1968 in the University of Münster, Germany at the Institute of Radiobiology by Professor Dr.Wolfgang Göhde.

  • First Published paper in Jan. 1969 “Dittrich W., Göhde W. "Impulsfluorometrie bei Einzelzellen in Suspensionen", Zeitschrift für Naturforschung (1969), Tübingen, Band 24 b, Vol. 3”

  • “Impulse Fluorometry” was the name used in the paper, “Impulse Cyto Photometry Technology” was called and still can be found in the Internet.

  • Hertzenberg started flow cytometry together with Bernhard Shaw (at that time V. P. of Becton Dickenson) in 1973, when the NIH had decided to develop an own American Flow Cytometer


More than 30 years of experience

More than 30 years of experience...

1969:

The ICP 11.

The first commercial flow Cytometer is born -the legendary ICP-11, developed in Münster and distributed by

Phywe AG, Göttingen.

Electronics size and fashion changed since that time...


Partec gmbh from m nster germany

1971:

ICP-11獲頒卓越產品獎

Award for outstanding product quality of theon the

Leipziger Herbstmesse.


Partec gmbh from m nster germany

需要判定細胞濃度及強弱螢光細胞的數量

一個光學高度計

一個磅秤

在入口處分發螢光棒

裝設一個紅光與綠光偵測及計數器

讓來客漁貫進入…

最後一個客人進入時資料也得到了

螢光顯微鏡 vs. 流式細胞儀

  • 主辦單位想由演奏會台上 數觀眾中男生及女生人數


Fluidics flow chamber

Fluidics & Flow Chamber


How it works

How it Works


Partec gmbh from m nster germany

Sheath fluid

Laminar Coaxial Flow

The Bernoulli Effect

鞘液流動方向

Lower pressure

Velocity Gradient

細胞之輸送

Hydrodynamic Focusing

Viscous drag along walls. Particles move to low pressure area


Partec gmbh from m nster germany

Laser

Light

雷射與螢光訊號

High angle scatter : SSC

大角度散射代表細胞內部

細胞質密度及表面顆粒性

Granularity

Low angle scatter : FSC

細胞顆粒大小

螢光散射

Fluorescence at longer wavelengths.

Intrinsic內部螢光

(自體螢光 auto fluorescence)

and extrinsic.


Partec gmbh from m nster germany

流式細胞儀之數據分析圖形


Partec gmbh from m nster germany

DNA Analysis

Cell Cycle

Cell Viability: Cell Dying by Apoptosis

Tumor Cell Analysis

Immunophenotyping

Simultaneous DNA content and Cell-Surface Immuno- fluorescence Analysis

Matching for Solid Organ Transplantation

Cell Membrane Potential Analysis

Sperm Chromatin Structure Assay

Intracellular pH

Intracellular Ionized Ca

Lysosomal Proton Pump Activity

“No-Wash” procedures for Nuclei Antigen Detection

Simultaneous Analysis of Cellular RNA & DNA content

PRINS & FISH

Enzyme Kinetics

Reticulocyte Analysis & Maturity Index

Cell Sorting

….

流式細胞儀部分應用


Partec gmbh from m nster germany

儀器操作步驟簡介


Light source

Light Source

Various light sources:

  • Argon

  • NdYAG

  • HeNe

  • Red Laser Diode

  • HBO UV lamp

  • HeCd


Partec gmbh from m nster germany

螢光發光原理及光譜


Sorting

Sorting 細胞分選原理

  • 1979:

  • The PAS-II.

  • mercury arc lamp (UV)

  • best DNA measurement precision (CV< 1%)

  • 3 optical parameters + electrical cell size

  • first piezo activated cell sorting (patented), real time acquisition

  • absolute cell counting

  • Motorola 6809, 68000 and 68020 processors

  • real potentiometers for PMT high voltage

  • acquisition rate 10000 cells/s

  • interchangeable filters


Cytopeia influx

Cytopeia InFlux

  • High Performance Sorter

  • High Speed Sorting up to 50,000/sec

  • Easy to Operate

  • Multiple Lasers

    • Blue 200mw 488

    • Green, Red

    • Solid State UV Laser

  • Highly Hygiene for Cell Culture by Disposal Sorting device


Partec gmbh from m nster germany

體積小

低耗能源

輸出功率高

啟動時間短不需暖機

使用壽命長

目前現有發展雷射 :

Blue 488 nm 200 mW

Red 635 nm 25 mW

Green NdYAG 532 nm 50 mW

Violet Laser

UV Laser

Many More to Come

雷射發展之趨勢 – 固態雷射


Cyflow

CyFlow

Compact

Highest Stability and Highest Precision

Power Connection: Regular 100/240 VAC or Car Battery (12 V)

Single Platform Volumetric Absolute Counting

Up to 5 Parameters (FSC, SSC, FL1, FL2, FL3)

Single Laser SystemUV/Violet, Blue, Green, and Red Solid State Lasers

New Level of Price / Performance Ratio


Partec gmbh from m nster germany

CyLab


Partec gmbh from m nster germany

CyFlow in Space - ISS


Cyflow ml

CyFlow ML

  • 桌上型設計

  • 可裝 3 組雷射及 UV 燈 (red, green, blue, violet lasers)

  • FloMax® Software

  •   16 參數 (FSC, 2 x SSC, FL-1 - FL-13)

  • 體積小 (size: ca. 56 cm x 65 cm x30cm)

  • Windows™ XP 作業系統

  •   New 16 bit 數位處理系統

  • 定體積絕對細胞計數功能True Volumetric Absolute Counting

  • 德國光學系統 超強螢光訊號靈敏度低於 100 MESF

  • FCS, SSC 可偵測粒子直徑 < 0.2 um


Cyflow space

CyFlow® Space

  • 桌上型設計

  • 雙固態雷射六色螢光系統: 20 mW @ 488 nm 25 mW @ 635 nm option

  • 6 色 Immunophenotyping: FITC + PE + PE-Cy5,PE-Cy5.5, PerCP, PI, ECD + PE-Cy7 + APC, APC-Cy5.5 + APC-Cy7

  • 德國光學系統 超強螢光訊號靈敏度低於 100 MESF

  • FCS, SSC 可偵測粒子直徑 < 0.2 um

  • 定體積絕對細胞計數功能True Volumetric Absolute Counting

  • 細胞分選系統 PPCS Particle and Cell Sorter option


Partec gmbh from m nster germany

模組化光學系統


Partec gmbh from m nster germany

偵測微細粒子之靈敏度

Supported by the use of a PMT for the FSC parameter, the small white blood cell platelets (PLT) can safely be discriminated from red blood cells (RBC) in the scatter plot.


Yeast analysis dna and lipids with double excitation

Yeast Analysis DNA and Lipids with Double Excitation

DAPI (DNA, FL4) and red staining (neutral lipids, FL3) of brewery yeasts after double excitation with HBO-lamp (UV) and argon ion laser (488 nm, 20 mW)


Bacteria analysis rfp in e coli

Bacteria Analysis RFP in E. coli

Measurement of RFP-expression (FL2: orange-red) of an E. coli population after excitation with an YAG-laser (523 nm, 50 mW): 40% of the E. coli population show the red fluorescence due to RFP-expression.


Microbiology analysis in food

Microbiology Analysis in Food

Bacteria in chocolate drink. left: control, right: contaminated sample.

The discrimination between “chocolate” particles and bacteria is based on spectral analysis in the two fluorescence channels.


Partec gmbh from m nster germany

定體積絕對細胞計數

The measurement volume is defined by mechanical measurement, rather than reference beads.


Partec gmbh from m nster germany

定體積絕對細胞計數原理

  • 不需添加標準濃度粒子試劑

  • volume:200 ± 10 µl, 流速軟體控制 0 - 120 uL/sec

  • coincidence losses: < 1% at analysis rates < 2000/s

  • 分析後可以任意圈選而得到不同區域細胞群之濃度


Partec gmbh from m nster germany

德國科技堅持 嚴選零件

Partec流式細胞儀使用與火星計畫 Sujourner 相同的馬達


Leukocytes

減白血液製品的Leukocytes數目

  • 機械式體積測量精確度高

  • 不需添加標準濃度粒子試劑

  • 可偵測濃度低至 < 0,01 Leukocytes / µl (= 10 / ml)

  • 以單一螢光參數 HBO-Lamp / ExciteDAPI

  • “Digital” counting due to bright CyStain staining

  • 方便快速


Cd4 cd8

CD4/CD8 單位體積之細胞數目

Routine concentration analysis on CD4/CD8 for HIV monitoring.

The concentration of subpopulations can as well be analyzed “offline”, after analysis.


Cd34 stem cells rare event

CD34 Stem Cells / Rare Event

Rare event concentration analysis on CD34+.


4 colors

多重螢光 4 colors 分析

  • Dual laser excitation: Argon and red laser diode.

  • MultiColor gating.

  • 4 color analysis.


Partec gmbh from m nster germany

多螢光譜線之重疊


Flomax

FloMax – 螢光校正補償

  • N-color cross-talk compensation.

  • Matrix-based: Compensation of any parameters, e.g. FL1-FL3.

  • Correction of compensation at any time: No need to rerun samples.


Partec gmbh from m nster germany

FloMax 人性化電腦介面


Flomax report

FloMax Report


Fcm application workshop

FCM Application Workshop


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