Human Disease and Immune Discovery

Equipment

In addition to our main instruments, HDID offers other tools to support your research. These include:

  • Flow Cytometry Data analysis workstation: High performance windows based computer with 64GB of RAM and newest version of Flowjo with all plugins and dependencies (R and Rtool + packages) required for unbiased analysis of multiparameter flow cytometry data.  Software suit is regularly updated and maintained, we also offer remote connection to the workstation and training in data analysis.
  • Biosafety Cabinet: For sterile manipulations and to protect users from potential biohazard exposure.
  • Temperature control shaker: For incubation and enzymatic digestion of solid tissues to generate single cell suspension suitable for downstream analysis.

We also offer access to number of validated antibodies which is an easy and economical way to start building large staining panels for your research project.

Symphony S6 from BD Biosciences

S6 cell sorter is capable to physically separate up to 6 unique cell population based on expression of up to 30 markers.

Configuration:

5 lasers, 30 fluorescence detectors

UV laser at 355nm, 100mW power, 8 detectors:

Detector name in configuration Common fluorochrome equivalent

Filters configuration

(LongPass mirror, filter)

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BUV395 380LP, 379/28
DAPI FVS440, AF350, Blue Live/Dead 410LP, 450/50
BUV496 450LP, 515/30
BUV563 550LP, 586/15
BUV615 600LP, 610/20
BUV661 635LP, 670/30
BUV737 690LP, 740/35
BUV805 770LP, 820/60

 

Violet laser at 408nm, 100mW power, 8 detectors:

Detector name in configuration Common fluorochrome equivalent

Filters configuration

(LongPass mirror, filter)

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BV421 Pacific Blue 410LP, 431/28

BV480

or

BV510

 

 

Aqua

450LP, 470/15

 

505LP, 525/50

BV480/BV510 possible as an alternative configuration
BV570 550LP, 586/15
BV605 600LP, 610/20
BV650 635LP, 670/30
BV711 690LP, 710/50
BV750 740LP, 750/30
BV786 BV785 750LP, 780/60

 

Blue laser at 488nm, 150mW power, 6 detectors:

Detector name in configuration Common fluorochrome equivalent

Filters configuration

(LongPass mirror, filter)

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BB515 FITC, AF488 505LP, 515/20
BB630 600LP, 610/20
BB660* 635LP, 670/30
BB700* PerCP-Cy5.5 690LP, 710/50
BB755* 730LP, 750/30
BB790* 770LP, 780/60

 

Yellow-Green laser at 561nm, 150mW power, 5 detectors:

Detector name in configuration Common fluorochrome equivalent

Filters configuration

(LongPass mirror, filter)

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PE 570LP, 586/15
PE-CF594 PE/Dazzle594 600LP, 610/20
PE-Cy5* 650LP, 670/30
PE-Cy5.5* 690LP, 710/50
PE-Cy7* 750LP, 780/60

 

Red laser at 638nm, 140mW power, 3 detectors:

Detector name in configuration Common fluorochrome equivalent

Filters configuration

(LongPass mirror, filter)

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APC AF647 650LP, 670/30
APC_R700 AF700, 690LP, 710/50
APC-Cy7 APC/Fire750, APC-H7 750LP, 780/60

 

Cytek Aurora

Aurora is a new type of flow cytometry analyzer; it works by measuring full emission spectrum from each laser (thus the name: spectral flow cytometer) and uses unmixing algorithm to calculate individual fluorochrome contribution within emission of all fluorochromes in flow cytometry panel of antibodies.

Practically it means less restriction in fluorochromes options, ability to combine fluorochromes with similar emission profile which at the end allow us to create bigger panels with great signal to noise ration.

With 64 individual detectors theoretical number of markers analyzed simultaneously on Aurora is 62, currently we are using 42 colors high quality and modular flow cytometry panel for blood samples on Aurora. Similar panels are in development for different tissues.

Configuration:

5 lasers, 64 detectors (channels)

UV laser at 355nm, 20mW, 16 channels covering spectrum from 365 to 829 nm.,

Violet laser at 405nm, 100mW, 16 channels from 420 to 829 nm.

Blue laser at 488nm, 50mW, 14 channels from 498 to 829 nm.

Yellow-Green laser at 561nm, 50mW, 14 channels from 567 to 829 nm.

Red laser at 640nm, 80mW, 8 channels from 652 to 829 nm.

Aurora is also equipped with plate loader which allows for automated runs of up to 96 samples at once.

New and very useful function on Aurora is volumetric sensor which allows for absolute quantification of any gated populations (counts per ul)

Good starting point for new Aurora users is Cytek website:

https://cytekbio.com/pages/aurora#tab-overview

Great tool for panel design for Aurora is full spectrum viewer:

https://spectrum.cytekbio.com/

And of course any additional questions related to Aurora will be answered by HDID core staff, we can also schedule more advanced training and address questions related to specific project.