Log In Sign up
The largest database of trusted experimental protocols

Aria3 fusion

Manufactured by BD
Visit Supplier

The BD ARIA3 Fusion is a flow cytometry instrument designed for cell analysis and sorting. It is capable of detecting and analyzing multiple parameters of individual cells within a sample.

Automatically generated - may contain errors

Lab products found in correlation

Aria 2, by BD (2 mentions) Epcam apc cy7, by BioLegend (2 mentions) Cd45 bv510, by BioLegend (2 mentions) Nk1.1 bv605, by BioLegend (2 mentions) Cd44 pe im7, by BioLegend (2 mentions) Tryple, by Thermo Fisher Scientific (2 mentions) Epcam fitc 9c4, by BioLegend (2 mentions) Cd90 pe dazzle thy1, by BioLegend (2 mentions) Rneasy microrna isolation kit, by Qiagen (2 mentions) Hiseq 4000, by Illumina (2 mentions) Rna 6000 pico kit, by Agilent Technologies (2 mentions)

4 protocols using aria3 fusion

1

Isolation of Murine and Human Intestinal Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols
After 4 days of murine and 7 days of human co-culture, Matrigel was disrupted and cells were collected into 15ml falcon tubes. For murine ILC1 cultures Matrigel disruption was not required, and cells were gently rinsed from the bottom of the plate using PBS+2%FCS. Samples were rinsed with PBS, then dissociated in TrypLe (Gibco) for 20mins at 37°C. The sorting buffer after this step contained DNAse (250µg/ml), EDTA (1μl/ml), and HEPES (1μl/ml) to maintain single epithelial cells and avoid clumping. Cells were titruated gently, centrifuged and resuspended in sorting buffer. Cells were then filtered (70µm), having pre-coated the filter with sorting buffer to minimize cell loss, and either rinsed with PBS for fixable Live/Dead staining (UV or nearInfraRed, Thermofisher), or stained with EpCAM, CD45, and the requisite combination of antibodies for the experiment, and analyzed (BD Fortessa) or sorted (BD ARIA3 Fusion & BD Aria 2 using BD FACS Diva 8.0.1 software). Isolation of murine IEC and ILC1 following co-culture was performed using EpCAM–APC Cy7 (G8.8, BioLegend), CD45-BV510 (30-F11, bioLegend), NK1.1 BV605 (PK136, BioLegend), CD44-PE (IM7, BioLegend). Isolation of human IEC, FB, and hILC1 used fixable Live/Dead-UV or Live/Dead-nIR CD45-eFluor450(HI30) Invitrogen, EpCAM-FITC (9C4; BioLegend), CD90-PE/Dazzle (Thy1; BioLegend)
Jowett G.M., Norman M.D., Yu T.T., Arévalo P.R., Hoogland D., Lust S.T., Read E., Hamrud E., Walters N.J., Niazi U., Chung M.W., Marciano D., Omer O.S., Zabinski T., Danovi D., Lord G.M., Hilborn J., Evans N.D., Dreiss C.A., Bozec L., Oommen O.P., Lorenz C.D., da Silva R.M., Neves J.F, & Gentleman E. (2020). ILC1 drive intestinal epithelial and matrix remodelling. Nature materials, 20(2), 250-259.
+ Open protocol
+ Expand
2

Isolation of Murine and Human Intestinal Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols
After 4 days of murine and 7 days of human co-culture, Matrigel was disrupted and cells were collected into 15ml falcon tubes. For murine ILC1 cultures Matrigel disruption was not required, and cells were gently rinsed from the bottom of the plate using PBS+2%FCS. Samples were rinsed with PBS, then dissociated in TrypLe (Gibco) for 20mins at 37°C. The sorting buffer after this step contained DNAse (250µg/ml), EDTA (1μl/ml), and HEPES (1μl/ml) to maintain single epithelial cells and avoid clumping. Cells were titruated gently, centrifuged and resuspended in sorting buffer. Cells were then filtered (70µm), having pre-coated the filter with sorting buffer to minimize cell loss, and either rinsed with PBS for fixable Live/Dead staining (UV or nearInfraRed, Thermofisher), or stained with EpCAM, CD45, and the requisite combination of antibodies for the experiment, and analyzed (BD Fortessa) or sorted (BD ARIA3 Fusion & BD Aria 2 using BD FACS Diva 8.0.1 software). Isolation of murine IEC and ILC1 following co-culture was performed using EpCAM–APC Cy7 (G8.8, BioLegend), CD45-BV510 (30-F11, bioLegend), NK1.1 BV605 (PK136, BioLegend), CD44-PE (IM7, BioLegend). Isolation of human IEC, FB, and hILC1 used fixable Live/Dead-UV or Live/Dead-nIR CD45-eFluor450(HI30) Invitrogen, EpCAM-FITC (9C4; BioLegend), CD90-PE/Dazzle (Thy1; BioLegend)
Jowett G.M., Norman M.D., Yu T.T., Arévalo P.R., Hoogland D., Lust S.T., Read E., Hamrud E., Walters N.J., Niazi U., Chung M.W., Marciano D., Omer O.S., Zabinski T., Danovi D., Lord G.M., Hilborn J., Evans N.D., Dreiss C.A., Bozec L., Oommen O.P., Lorenz C.D., da Silva R.M., Neves J.F, & Gentleman E. (2020). ILC1 drive intestinal epithelial and matrix remodelling. Nature materials, 20(2), 250-259.
+ Open protocol
+ Expand
3

Isolation and RNA-seq of Cell Subsets

Check if the same lab product or an alternative is used in the 5 most similar protocols
The cells were harvested as described above for sorting by flow cytometry (BD ARIA3 Fusion using BD FACS Diva 8.0.1 software) into RLT (Qiagen) lysis buffer. RNA was harvested using RNeasy MicroRNA isolation kit (QIAGEN), and RIN values were assessed using RNA 6000 Pico Kit (Agilent). The library was prepared using SMARTSeq2 and sequenced by Illumina HiSeq 4000 at the Wellcome Trust Oxford Genomics Core, where basic alignment (GRCm38.ERCC (2011)) and QC was also performed.
Jowett G.M., Norman M.D., Yu T.T., Arévalo P.R., Hoogland D., Lust S.T., Read E., Hamrud E., Walters N.J., Niazi U., Chung M.W., Marciano D., Omer O.S., Zabinski T., Danovi D., Lord G.M., Hilborn J., Evans N.D., Dreiss C.A., Bozec L., Oommen O.P., Lorenz C.D., da Silva R.M., Neves J.F, & Gentleman E. (2020). ILC1 drive intestinal epithelial and matrix remodelling. Nature materials, 20(2), 250-259.
+ Open protocol
+ Expand
4

Isolation and RNA-seq of Cell Subsets

Check if the same lab product or an alternative is used in the 5 most similar protocols
The cells were harvested as described above for sorting by flow cytometry (BD ARIA3 Fusion using BD FACS Diva 8.0.1 software) into RLT (Qiagen) lysis buffer. RNA was harvested using RNeasy MicroRNA isolation kit (QIAGEN), and RIN values were assessed using RNA 6000 Pico Kit (Agilent). The library was prepared using SMARTSeq2 and sequenced by Illumina HiSeq 4000 at the Wellcome Trust Oxford Genomics Core, where basic alignment (GRCm38.ERCC (2011)) and QC was also performed.
Jowett G.M., Norman M.D., Yu T.T., Arévalo P.R., Hoogland D., Lust S.T., Read E., Hamrud E., Walters N.J., Niazi U., Chung M.W., Marciano D., Omer O.S., Zabinski T., Danovi D., Lord G.M., Hilborn J., Evans N.D., Dreiss C.A., Bozec L., Oommen O.P., Lorenz C.D., da Silva R.M., Neves J.F, & Gentleman E. (2020). ILC1 drive intestinal epithelial and matrix remodelling. Nature materials, 20(2), 250-259.
+ Open protocol
+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required

Sign up now

Revolutionizing how scientists
search and build protocols!