Preparation method suitable for mandulapalka flow cytometry sample and cell lysis solution

A cell lysate and flow cytometry technology, which is applied to the preparation of oily bean flow cytometry samples and the field of cell lysate, can solve the problems of stray peaks, small number of single cell nuclei, incomplete flow detection results, etc. To achieve the effect of improving detection accuracy, increasing the number of single cell nuclei, and improving accuracy

Pending Publication Date: 2022-04-05
INST OF IND CROPS HENAN ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In the previous research, it was found that using conventional lysates and method procedures, there are often a large number of cell debris and impurities, and the number of single cell nuclei is small and incomplete, resulting in miscellaneous peaks or no target peaks in flow cytometry results.

Method used

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  • Preparation method suitable for mandulapalka flow cytometry sample and cell lysis solution
  • Preparation method suitable for mandulapalka flow cytometry sample and cell lysis solution
  • Preparation method suitable for mandulapalka flow cytometry sample and cell lysis solution

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Embodiment 1

[0036] This embodiment provides a method for preparing samples suitable for flow cytometry of oily safflower, including the following steps:

[0037] (1) Select the stem tiller of Yuyousha No. 2 (round-grain type) plant with a total number of leaves of 5, and use scissors to cut young leaves of 2 cm 2 , Rinse 2-3 times with distilled water to remove the dust on the surface of the leaves, and dry the residual water with absorbent paper;

[0038] (2) Put the leaves into a petri dish with a diameter of 5.5 cm containing 400 ul of pre-cooled cell lysis buffer, and cut the leaves vertically for 60 seconds with a sharp double-sided blade to obtain a leaf slurry; the whole process material is immersed in in cell lysis buffer and on ice;

[0039] (3) Add 400 ul of pre-cooled cell lysate to the leaf slurry prepared in step (2), blow gently with a pipette 8 times, mix well, and then let stand on an ice box for 15 minutes;

[0040] (4) Put a 30 μm filter on a 1.5ml centrifuge tube, use a...

Embodiment 2

[0047] This embodiment provides a method for preparing samples suitable for flow cytometry of oily safflower, including the following steps:

[0048] (1) Select the stem tiller of Yuyousha No. 3 (long-grain type) plant with a total number of leaves of 5, and use scissors to cut young leaves of 2 cm 2 , Rinse 2-3 times with distilled water to remove the dust on the surface of the leaves, and dry the residual water with absorbent paper;

[0049] (2) Put the leaves into a petri dish with a diameter of 5.5 cm containing 400 ul of pre-cooled cell lysis buffer, and use a sharp double-sided blade to cut the leaves vertically for 50 seconds quickly to obtain a leaf slurry; the whole process material is immersed in in cell lysis buffer and on ice;

[0050] (3) Add 400 ul of pre-cooled cell lysate to the leaf slurry prepared in step (2), blow gently 6 times with a pipette gun, mix well, and then let stand on the ice box for 15 minutes;

[0051] (4) Put a 30 μm filter on a 1.5ml centri...

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Abstract

The invention relates to the field of plant flow cytometry detection, in particular to a preparation method of a flow cytometry sample suitable for cyperus esculentus and a cell lysis solution. According to the preparation method, the cell lysis solution is added twice in the leaf slurry preparation process, so that the cell lysis effect is ensured, the number of obtained single cell nucleuses and the integrity of the cell nucleuses are improved, impure peaks detected and analyzed by a flow cytometer are reduced, and the detection accuracy is improved. Meanwhile, the improved cell lysis buffer solution suitable for preparation of the cyperus esculentus flow cytometry sample provided by the invention is used for treating a tissue sample, and all the components have a synergistic effect in a limited dosage range, so that complete cell nucleuses can be effectively released, residual cytoplasm fragments can be removed, and the influence of a phenol mixture on a result is reduced; the stability of cell nucleuses in the suspension liquid is kept, the agglutination is prevented, and the accuracy of flow cytometry analysis and detection is improved.

Description

technical field [0001] The invention relates to the field of plant flow cytometry detection, in particular to a preparation method and a cell lysate suitable for flow cytometry samples of oily bean. Background technique [0002] Cyperus esculentus L., also known as Cyperus esculentus L., enjoys the reputation of "fruit of life". The sedge is a plant of the Cyperaceae Cyperus genus, which can bloom under certain conditions. The aboveground part grows stems and leaves, and the underground stolons expand into beans, and the stem beans are used for oil extraction or flour milling. The oilseed bean is cold-resistant and drought-resistant, and suitable for cultivation on sandy soil and other marginal lands. It is an oil crop with high economic value. The development of the oily bean industry is of great significance in ensuring national grain and oil security. As a vegetative plant, there are often phenomena such as chromosome doubling, centromere breakage, fusion, etc. in nature...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N1/28G01N1/30G01N15/14C12N5/04
Inventor 李春鑫王会伟王艳张向歌李居政王树峰宋文栩
Owner INST OF IND CROPS HENAN ACAD OF AGRI SCI
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