招生          学生培养           合作伙伴      
  •   细胞分选平台


细胞分选平台
    平台简介应用&服务平台成员招聘信息

Introduction

The cell sorting core at SIAIS provides flow cytometric analysis and cell sorting for ShanghaiTech University researchers and the surrounding biotech and biomedical research community. This essential resource aids investigators performing research in antibody drugs development, cellular immunology, cancer related areas and stem cell biology. Samples are prepared by investigators and then delivered to the Core for flow cytometric analysis and/or cell sorting.
Flow cytometry-Cell analysis

Flow cytometry is a highly sensitive technology for single cell fluorescent signals measurements. By using specific fluorescent probes, this technology allows simultaneous multiparametric analysis of many thousands of cells per second, enabling researchers to rapidly analyze complex cell populations.
Cells or particles labeled with fluorescent molecules enter a fluid stream and flow at a constant speed in flow cytometer. When the cells pass through laser focuses along the stream one by one, the fluorescent probes are excited by the laser of specific wavelength and then emit light. The optics collects and transduces fluorescent light to the detector according to its color. Then the fluorescent light is detected, amplified and translated into an electronic signal, which will be processed by signal electronics and sent to a computer for presentation. Information about the cell is recorded and the result can be visually displayed on the computer screen in real time.

Flow cytometry-Cell Sorting

Cell sorting is the physical isolation and enrichment of selected cells. Sorting on a flow cytometer is executed just after the standard measuring process. During sorting process the stream is controllably vibrated at a specific frequency to stably break it into droplets. These droplets containing cells with selected values by investigator are then positively or negatively charged more or less and go through a constant electric field. As a result the charged droplets are sent along selected trajectories into vessels (tube or plate), and the uncharged fluid containing non-target cells flow into waste drawer.

When should you use flow cytometric cell sorting?

1). To enrich target cell population at a very high purity of 95%-100%.
2). To sort desired cells on the basis of multiple parameter measurements.
3). To separate cell populations with some low expression antigens on cell surface.
4). To isolate cells according to the accurately quantitative density of specific cell markers.
5). To acquire one single cell in multi-well plates.
6). To separate cells on the basis of internal constituents or functional staining, e.g. fluorescent hydrolysates.
7). To detect and sort very rare cells (0.001% or less) from mixed populations.

Instrument

BD FACSAriaIII for analysis and sorting

Laser
Detector
Mirror
BP
Fluorochromes Detected
Blue
488nm/
13mW
A
655LP
695/40
PerCP-Cy5.5, PE-Cy5.5, 7AAD, PE-Alexa700
B

502LP
530/30
FITC, Alexa488, YFP, GFP, CFSE
Mitotracker FM, Rhodamine 123,
Calcein, DCF, DCFDA, CMFDA
C
488/10
SSC
Yellow
561nm
/30mW
A
735LP
780/60
PE-Cy7
B
685LP
710/50
PE-Cy5.5, PE-Alexa 700
C
630LP
670/14
PE-Cy5,
D
600LP
610/20
PE-Texas Red, DsRed, RFP, mCherry,
Mitotracker Red, FP635/Far Red, TMRM, PI
E
582/15
PE, Pyronin Y, Tomato, Orange,
Alexa594, Cy3, PKH26, CMRA
Red
633nm
/11mW
A
755LP
780/60
APC-Cy7, APC-Alexa750, APC-H7,
APC-Alexa780, APC-eFluor780
B
690LP
735/45
Alexa700
C
502LP
660/20
APC, Alexa 647, Mitoxantrone, Cy5
Violet 407nm
(10mW)
/375nm(5mW)
A
502LP
510/50
Pacific Orange, Live/Dead Aqua,
BV510, Cascade Yellow , Amcyan
B
450/40

DAPI, Hoechst, BV421, eFluor450,
Pacific Blue, CFP, BFP, Live/Dead
Violet, Cascade Blue

Quality control: Run BD CS&T Procedure every 48hour.




 
地址
|

上海张江高科技园区海科路99号6号楼 邮编:201210 
邮箱: siais@shanghaitech.edu.cn

 

Copyright © 2015 上海科技大学免疫化学研究所 版权所有