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About us Medical Sciences Research Flow Cytometry Unit Instruments
University Medical Center Groningen

Beckman Coulter MoFlo Astrios

Beckman Coulter MoFlo Astrios
Beckman Coulter MoFlo Astrios

General Information

In the FCU (Flow Cytometry Unit) one MoFlo Astrios sorter is located. This high speed sorter is used for research purposes.

The MoFlo Astrios is a high speed sorter equipped with 7 lasers (355 nm, 405 nm, 488 nm, 532 nm, 561 nm, 592 nm and 640 nm The MoFlo Astrios provides the ability to analyze or sort up to 32 parameters (FSC, 6 SSC and 25 colors). According to the technical specification it can achieve a validated sort rate of up to 70.000 events per second . In our practice, sorting rates till 25.000 events per second can be achieved with good recovery, yield and viability using the standard 70 nozzle. Higher speeds (45.000 cells/sec) are very well possible but will lead to a lower yield (because of Poisson statistics). Six different populations can be sorted at the same time. Each population can be sorted in its own sort mode (Single, Purify and Enrich)

Optional nozzles sizes (100 um, 120 um and 150 um) can be used besides the standard 70 nozzle, to sort bigger cells (This prevents clogging of the nozzle and creates better side streams). On the backside it will reduce the speed a lot (max 8000 cells/sec with a 100 um nozzle) but because of lower shear stress involved it is more gentle for the cells.

Multiple tube formats can be applied for sorting using the temperature controlled SmartSampler. The sorted cells can be collected in multiple tube formats like eppendorf cups, 5, 15 and 50 ml tubes. The Cyclone enables (single cell) sorting in different plates (12, 24, 48, 60 (Terasaki), 96, 384 wells) and also on slides (customable format).

Samples are acquired (32 bits digitalization in 5 decades) using the Summit software. Samples are saved as FCS3.0 files. Compensation afterwards is possible.

Sample preparation

Before sorting the sample needs to be filtered, to eliminated cell aggregrates, using the BD Falcon 5 ml polystyrene Round-Bottom Tube with Cell-Strainer Cap (REF 352235) (35 um). Doing this prevents clogging in the sample line or nozzle and results in a better, more time-efficient way of sorting.

Specifications

Laser

Laser

Wavelength

Optical Power (at the Nozzle)

Ultraviolet 355 nm 20 mW
Violet 405 nm 30 mW
Blue 488 nm 125 mW
Green 532 nm 95 mW
Yellow Green 561 nm 150 mW
Orange 592 nm 150 mW
Red 642 nm 80 mW

Standard Configuration

355 nm Laser

Filter

Fluorochrome

379/21

448/59

DAPI, Hoechst Blue

692/75

Hoechst Red

405 nm Laser

Laser

Filter

Fluorochrome

Violet 405 nm

405/10

SSC Violet Laser

448/59

Pacific Blue, H450, E450, Brilliant Violet 421

546/20

Pacific Orange

607/35

eFluor 600(605), BV605

795/70

488 nm Laser

Laser

Filter

Fluorochrome

Blue 488 nm

488/6

SSC Blue Laser

513/26

FITC, Alexa 488, CFSE, BB515

576/21

PE**, dsRed**, dTomato**, PI**, mBanana**

620/29

PE-Texas Red, PI**

664/22

PE-Cy5, PI**, PerCP

710/45

PE-Cy5.5, PerCP-Cy5.5

795/70

PE-Cy7

532 nm Laser

Laser

Filter

Fluorochrome

Green 532 nm

530/11

SSC Green Laser

576/21

PE**, dsRed**, dTomato**, mOrange, mBanana*

622/22

PI*

664/22

PE-Cy5

692/18

PE-Cy5.5

736/47

561 nm Laser

Laser

Filter

Fluorochrome

Yellow/Green 561 nm

561/14

SSC Yellow Laser

579/16

PE*, dsRed*, dTomato*

614/20

mCherry, mRaspberry**

692/75

PE-Cy5.5

795/70(from the Blue Laser)

592 nm Laser

Laser

Filter

Fluorochrome

Orange 592 nm

592/8

SSC Orange Laser

620/29

mPlum, mCherry, mRaspberry*

671/30

722/44

795/70

642 nm Laser

Laser

Filter

Fluorochrome

Red 640 nm

642/10

SSC Red Laser

671/30

APC, A647

`

722/44

A700

795/70

APC-Cy7, APC-A750, APC-EFluor 780

*Best choice

** Second best choice

Sample Formats

Sample format(s)

Smartsampler

0.5 ml, 1.0 ml, 1.5 ml, 5 ml, 7 ml, 15 ml, 50 ml tubes

Sorted Cell Collection Formats

Tube Holder

Sample format(s)

Plate deposition

6-1536- well plates, 60 well Terasaki plate

Tube deposition

5 mL tube holder

6x 1.5 or 5 mL tube sizes

15 ml tube holder

2x 15 mL tubes

50 ml tube holder

2x 50 mL tubes

50 mL & 5 mL holder

3x 5 mL plus 1x 50 mL tubes

Slide Deposition

Custum slide configurations

Last modified:15 October 2019 1.23 p.m.