A method for detection of PD-1 in T cells based on the quality assessment of cellular mitochondria
A detection method, PD-1 technology, applied in measurement devices, material excitation analysis, fluorescence/phosphorescence, etc., can solve the problems of deviation of detection results, inability to obtain accurate solid tissue, etc.
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no. 1 example
[0041] A method for detecting PD-1 in T cells based on cell mitochondrial quality assessment, comprising the following steps:
[0042] S10: Obtaining T cells; including:
[0043] S11: Using lymphocyte separation medium to extract and separate human peripheral blood samples to obtain PBMCs;
[0044] S12: adding antibody-coated magnetic beads to the PBMC, and incubating at room temperature for 20 minutes to obtain labeled T cells;
[0045] S13: adding a buffer to the labeled T cells for washing, then adding an elution buffer, and incubating at 30° C. for 20 minutes; finally washing with the buffer to obtain the T cells.
[0046] Wherein, the buffer is PBS buffer containing 1% FBS; and the elution buffer contains 300U / mL chymopapain.
[0047] S20: adding CD4 PE Cyanine7 and PD-1 PE antibodies to the T cells to obtain sample 1;
[0048] S30: adding a mitochondrial probe to the sample 1, incubating at 37° C. for 30 minutes, using the mitochondrial fluorescent probe as a cell met...
no. 2 example
[0052] In this embodiment, the expression level of PD-1 in infected samples is detected, and the specific steps are as follows:
[0053] S10: Obtaining T cells; including:
[0054] S11: Using lymphocyte separation medium to extract and separate human peripheral blood samples to obtain PBMCs;
[0055] S12: adding antibody-coated magnetic beads to the PBMC, and incubating at room temperature for 30 minutes to obtain labeled T cells;
[0056] S13: adding a buffer to the labeled T cells for washing, then adding an elution buffer, and incubating at 37° C. for 30 minutes; finally washing with the buffer to obtain the T cells.
[0057] Wherein, the buffer is PBS buffer containing 1% FBS; and the elution buffer contains 100 U / mL chymopapain.
[0058] S20: adding CD4 PE Cyanine7, CD8 FITC and PD-1 PE antibodies to the T cells to obtain sample 1;
[0059] S30: adding a mitochondrial probe to the sample 1, incubating at 37° C. for 30 minutes, using the mitochondrial fluorescent probe ...
no. 3 example
[0063] In this embodiment, the expression level of PD-1 in tumor samples is detected, and the specific steps are as follows:
[0064] S10: Obtaining T cells; including:
[0065] S11: Using lymphocyte separation medium to extract and separate human peripheral blood samples to obtain PBMCs;
[0066] S12: adding antibody-coated magnetic beads to the PBMC, and incubating at room temperature for 30 minutes to obtain labeled T cells;
[0067] S13: adding a buffer to the labeled T cells for washing, then adding an elution buffer, and incubating at 37° C. for 30 minutes; finally washing with the buffer to obtain the T cells.
[0068] Wherein, the buffer is PBS buffer containing 1% FBS; and the elution buffer contains 100 U / mL chymopapain.
[0069] S20: adding CD4 PE Cyanine7, CD8 FITC and PD-1 PE antibodies to the T cells to obtain sample 1;
[0070] S30: adding a mitochondrial probe to the sample 1, incubating at 37° C. for 30 minutes, using the mitochondrial fluorescent probe as ...
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