Amnis Imagestream-X imaging flow cytometer

Room: S633
Contact: Ewa.Menet@yale.edu or (203)785-7958

Manufacturer's Website

Amnis Imagestream-X imaging flow cytometer provides users with the ability to gain detailed images of a large number of cells in a relatively short period of time and with the opportunity to perform a range of novel applications including co-localization, internalization, stem cell differentiation, and cell-cell interactions.

ImageStreamX produces up to 12 high resolution images of each cell directly in flow, at rates exceeding 1,000 cells per second, and with the fluorescence sensitivity of conventional flow cytometers. These capabilities allow you to quantitate cellular morphology and the intensity and location of fluorescent probes on, in, or between cells, even in rare sub-populations and highly heterogeneous samples.

With the ImageStreamX you can:

  • Image cells directly in suspension with the resolution of a 40X microscope and the fluorescence sensitivity of flow cytometers
  • Analyze highly heterogeneous samples and rare cell sub-populations at speeds exceeding 1,000 cells per second
  • Perform phenotypic and functional studies at the same time using up to five lasers and 12 images per cell
  • Quantitate virtually anything you can see using the IDEAS® software package’s numerous pre-defined fluorescence and morphologic parameters

ImageStreamX Optics

ImageStreamX has 4 lasers (488 nm blue laser, 561 nm green laser, 635 nm red laser, 405 nm violet laser and a 785 nm laser used by SSC). The output of each laser can be independently adjusted from 10 to 100 percent of maximum laser power for greater experimental flexibility and optimal results.

ImageStreamX Fluorochrome Chart – 2 camera STD

Camera 1

Excitation Laser (nm)

Ch

Band(nm)

405

488

561

658

785

1

430-480

BRIGHTFIELD

2

480-560

FITC, AF488, GFP, YFP, DyLight488, PKH67, Syto 13, Spectrum Green, Lyso TrackerGreen, Mito TrackerGreen, QD525*

3

560-595

PE, PKH26, Cy3, AF555, DSRed

PE, Cy3, AF546, AF555, DyLight549, DyLight594*, PKH26, DSRed, SpectrumOrange, Mito TrackerOrange

4

595-660

PE-TexRed*, ECD*, PE-AF610*, 7AAD*, PI*, RFP, QD625*, eFluor625*

AF568*, AF594*, AF610*, DyLight594*, PE-TexRed*, ECD*, TexRed*, PE-AF610*, RFP, mCherry*, 7AAD*, PI*

5

660-745

PE-Cy5*, PE-AF647*, PerCP*, Per-CP-Cy5.5*, DRAQ5*, QD705*, eFluor650*

PE-Cy5*, PE-AF647*, DRAQ5*

6

745-800

PE-Cy7*, PE-AF750*, QD800

PE-Cy7*, PE-AF750*,

SSC

Camera 2

Excitation Laser (nm)

Ch

Band(nm)

405

488

561

658

785

7

430-505

DAPI, Hoechst, PacBlue, CascadeBlue, AF405, eFluor405, DyLight405, CFP, LIVE/DEAD Violet

8

505-570

PacOrange, Cascade Yellow, AF430, QD525*

9

570-595

BRIGHTFIELD

10

595-660

QD625*, eFluor625*

11

660-745

QD705*, eFluor650*

AF647, AF660, AF680, DRAQ5*, APC, Cy5, DyLight649, DyLight680, PE-AF647*, PE-Cy5*, PerCP*, PerCP-Cy5.5*

12

745-800

QD800*

APC-Cy7, APC-AF750, APC-eFluor750, Cy7, AF750, DyLight750, eFluor750, PE-Cy7*, PE-AF750*

Dyes with an* are excited by one laser directed to camera 1 and another laser directed to camera 2. The channel that the dye will appear brightest in depends on the relative laser power used. Recommended dyes (based on excitation and detection channels) are in boldface. QD565 and QD585 are not included because their primary fluorescence appears in the Ch9 Brightfield.