Novel Barcoding 89Y reagent for mass cytometry

A technology of flow cytometry and reagents, applied in the field of new Barcoding89Y reagents, to achieve the effect of increasing screening work

Inactive Publication Date: 2017-12-22
浙江普罗亭健康科技有限公司
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  • Abstract
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  • Claims
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  • Novel Barcoding 89Y reagent for mass cytometry

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Embodiment 1

[0021] 1. Prepare reagents:

[0022] a, hydrochloric acid (AR grade, 10011018, Sinopharm Chemical Reagent Co., Ltd.),

[0023] b. Yttrium (III) chloride hexahydrate (204919, Sigma),

[0024] c, X8 polymer, L buffer ( X8 Multimetal Labeling Kit—40 Rxn, 201300, Fluidigm),

[0025] d, phosphate buffer (pH7.4, Gibco),

[0026] e, 104Pd reagent (S00114, Fluidigm).

[0027] 2. Prepare 1N hydrochloric acid solution with double distilled water.

[0028] 3. Prepare 100mM yttrium(III) chloride hexahydrate solution with 1N hydrochloric acid solution, and mix thoroughly.

[0029] 4. Take out a tube of X8polymer, add 97.5μL L buffer, pipette evenly to fully dissolve X8polymer.

[0030] 5. In step 4, add 2.5 μL of 100 mM yttrium (III) chloride hexahydrate solution, mix well and incubate at 37° C. for 30 min.

[0031] 6. After the incubation, transfer the entire solution to a 3kd ultrafiltration tube, and centrifuge at 12000g for 25min at room temperature. Discard the supernatant, c...

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Abstract

The invention discloses a novel Barcoding 89Y reagent for mass cytometry. The reagent is prepared through the following steps: firstly, adding 39-195 mu L L of buffer to a tube of X8 polymer, blowing and beating the mixture uniformly to fully dissolve the X8 polymer, then adding 1-5 mu L of a 100 mM yttrium (III) chloride hexahydrate solution, and performing uniform stirring and incubation; secondly, transferring all the solution to a 3 kd ultrafiltration tube after incubation, performing centrifugation, removing a supernatant, collecting a liquid in the ultrafilter tube to a centrifugal tube, cleaning the ultrafiltration tube with 40-200 mu L L of buffer, collecting all the liquid in the centrifugal tube, and fully mixing the liquid uniformly to obtain the Barcoding 89Y reagent. Cells can be labeled, and accordingly, screening work of increased flux of flow cytometry can be performed, and the barcoding channel applicable to mass cytometry is extended.

Description

technical field [0001] The invention relates to the technical field of cell biology, in particular to a novel Barcoding 89Y reagent for mass spectrometry flow cytometry. Background technique [0002] Barcoding technology is an important application in traditional fluorescent flow cytometry. Its principle is to use multiple labels to label cells, and to distinguish markers according to different combination types, so that flow cytometry can perform screening work with increased throughput. Barcoding technology has many advantages: reduce antibody loss, shorten sample collection time, reduce cross-contamination during sample transfer, eliminate sample-to-sample (tube-to-tube) differences during staining, remove cell dimers and cell debris, etc. [0003] After the birth of mass spectrometry, this technology has been further developed. Mass cytometry can provide more channels for Barcording labeling, so higher throughput can be achieved. At present, there are only the followin...

Claims

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Application Information

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IPC IPC(8): G01N15/14
CPCG01N15/14
Inventor 王甜
Owner 浙江普罗亭健康科技有限公司
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