Isotonic azide free solution

Peripheral venous blood samples were collected between 9 a.m. and 10 a.m. before the questionnaires were filled in order to control for diurnal variation. Patients’ cellular immunity tests were completed by the professional staff working at the clinical laboratory of the hospital. T cell subsets, including the percentage of total T cells (CD3+), helper T cells (CD4+), cytotoxic T cells (CD8+), NK cells (CD56+), and B cells (CD19+), were measured. Flow cytometry was used to assess T and NK cell counts with a Cytomics™ FC500 series instrument from Beckman Coulter (USA). Reagents from BD Bio-Engineering Co., Ltd. were used. Cells were fixed in 3% formaldehyde in an isotonic azide-free solution (Beckman Coulter, Luton, UK). Labeled antibodies were added at the recommended concentrations and then cells were washed. Cells were cultured in the dark, and then excess antibodies were washed out.

To control diurnal variations, peripheral venous blood samples were collected between 9 am and 10 am before the questionnaires were completed. Patients’ cellular immunity tests were completed by the professional staff working at the clinical laboratory of the hospital. The immunity tests evaluated T-cell subsets, including the percentage of total T cells (CD3⁺), helper T cells (CD4⁺), cytotoxic T cells (CD8⁺), NK cells (CD56⁺), and B cells (CD19⁺). Flow cytometry was used to assess T- and NK-cell counts with a Cytomics™ FC 500 series instrument from Beckman Coulter (Brea, CA, USA). Reagents from BD Bio-Engineering Co., Ltd. (Franklin Lakes, NJ, USA), were used. Cells were fixed in 3% formaldehyde in an isotonic azide-free solution (Beckman Coulter). Labeled antibodies were added at the recommended concentrations, and then, cells were washed. Cells were cultured in the dark, and then, excess antibodies were washed out.

To control for diurnal variation, peripheral venous blood samples were collected between 9 am and 10 am before the questionnaires were filled. Patients’ cellular immunity tests were completed by the professional staff working at the clinical laboratory at Anshan Cancer Hospital. They evaluated T-cell subsets including the percentage of total T cells (CD3+), helper T cells (CD4+), cytotoxic T cells (CD8+), NK cells (CD56+), and B cells (CD19+). Flow cytometry was used to assess T- and NK-cell counts with a Cytomics™ FC500 series instrument from Beckman Coulter (Brea, CA, USA). Reagents from BD Bio-Engineering Co., Ltd were used. Cells were fixed in 3% formaldehyde in an isotonic azide-free solution (Beckman Coulter). Labeled antibodies were added at the recommended concentrations and then cells were washed. Cells were cultured in dark, and then excess antibodies were washed out.