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University Medical Center Groningen

Beckman Coulter MoFlo XDP

Beckman Coulter MoFlo XDP
Beckman Coulter MoFlo XDP

General Information

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

The MoFlo XDP is a high speed sorter with 4 lasers (488 nm, 532 nm, 404 nm and 642 nm Lasers. The MoFlo XDP provide the ability to analyze/sort up to 13 parameters (FSC, SSC and 11 colors). According to the technical specification it can achieve a validated sort rates of up to 70.000 events per second . In our practice, sorting rates till 25.000 events per second are achieved with good recovery, yield and viability using the standard 70 nozzle. Higher speeds (30.000-35.000 events/sec) are very well possible but will lead to lower yield (because of Poisson statistics). 4 different populations can be sorted at the same. 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 the 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 tubes and 5 ml tubes. The Cyclone enables sorting in different plateformats (6, 12, 24, 48,60 (Terasaki) 96 and 384 wells) and also on slides. Samples are acquired (32 bits – 5 decades) using the Summit software. Samples are saved 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)

Violet (Coherent Cube)

404 nm

50 mW

Blue (Coherent Sapphire Laser)

488 nm

125 mW

Green (Coherent Sapphire Laser)

532 nm

75 mW

Red (Coherent Cube)

647 nm

60 mW

Standard Configuration

488/532 nm Laser (Co-linear on one pinhole)

Detector

Filter

Fluorochrome

FSC

-

SSC

488/6

-

FL - 1

513/17

Alexa Fluor 488, FITC, GFP

FL - 2

575/25

PE

FL - 3

610/20

PI, PE-CF594, PE-Texas Red, PE-eFluor 610

FL - 4

684/24

PE-Cy5, PE-A647, PE-Cy5.5, PerCP

FL - 5

720/13

PE-A700

FL - 6

785/62

PE-Cy7

405/635 nm laser (Co-linear on one pinhole)

Detector

Filter

Fluorochrome

FL - 13

795/68

APC-Cy7, APC-H7, APC-eFluor 780

FL - 14

671/28

APC, Alexa Fluor 647, Cy5, eFluor 660

FL - 15

722/42

Alexa Fluor 700

FL - 16

447/60

Pacific Blue, BV421, Horizon V450, eFluor 450,

FL - 17

520/38

Pacific Orange, BV510, Horizon V500, Pacific Orange

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

Sample format(s)

Plate/Slide deposition (Cyclone)

6-1536- well plates, 60 well Terasaki and custum slide configurations

Tube deposition 5 mL tube holder

1.5, 5 mL tube sizes

Last modified:15 October 2019 1.24 p.m.