相关实验视频

Updated: Jun 9, 2025

Multicolor Flow Cytometry-based Quantification of Mitochondria and Lysosomes in T Cells

Multicolor Flow Cytometry-based Quantification of Mitochondria and Lysosomes in T Cells

Published on: January 9, 2019

试剂定位在流细胞计中的作用力

Diana L Bonilla¹, Alberta Paul¹, Jesus Gil-Pulido²

¹Scientific Commercialization, Cytek Biosciences, Inc., 47215 Lakeview Boulevard, Fremont, CA 94538, USA.

|October 25, 2024

概括

通过定位优化试剂度对于精确的流动细胞计量至关重要. 适当的抗体定位通过最大限度地增加信号和最大限度地减少背景噪声来确保可靠的结果,从而实现精确的细胞分析.

科学领域:

免疫学免疫学免疫学
细胞生物学细胞生物学
生物技术是生物技术.

背景情况:

流细胞计能够对多个细胞参数进行高分辨率,高通量分析.
精确的流量细胞计数据依赖于优化的试剂度,使试剂验证至关重要.
抗体定位是测试优化的关键第一步,以确定最佳试剂度.

研究的目的:

总结流细胞计中的抗体定位的基本原理和最佳实践.
评估各种实验条件如何影响最佳标位选择和种群分辨率.
强调抗体定位的必要性,以获得可靠和可重复的流细胞计结果.

主要方法:

对抗体的连续稀释在表达目标抗原的细胞上进行了测试.
通过绘制积极细胞和光强度与负面事件的图形来生成度-反应曲线.
通过计算最高的信号与噪音比率来确定最佳抗体度.

主要成果:

低于最佳的抗体度 (过低或过高) 会导致数据不准确,高可变性和信号问题.
最佳的定位确保了最大的信号,结合点的和和最小的抗体过剩.
仪器,样品,染色,采集和分析条件等实验变量可以影响最佳标位选择.

结论:

抗体定位是实现高数据分辨率和流细胞计可靠结果的必不可少的.
每种样品类型,试剂批量和实验条件都需要特定的定位以获得最佳性能.
坚持定位最佳实践可以提高免疫学评估的准确性和可重复性.

关键词:

抗体定位抗体定位数据可复制性数据可复制性流动细胞计量是流动细胞计量的方法.频谱细胞计量 (Spectral Cytometry) 是一种检测方法.

更多相关视频

Discrimination of Seven Immune Cell Subsets by Two-fluorochrome Flow Cytometry

Discrimination of Seven Immune Cell Subsets by Two-fluorochrome Flow Cytometry

Published on: March 5, 2019

Quality-Controlled Sputum Analysis by Flow Cytometry

Quality-Controlled Sputum Analysis by Flow Cytometry

Published on: August 9, 2021

相关概念视频

Flow Cytometry

Flow Cytometry

The development of flow cytometry techniques began in 1934 with initial attempts by Andrew Moldavan, a bacteriologist who counted the cells in a flowing capillary system. Moldavan pumped cells through a capillary tube focused under a microscope for visualization. The invention of photometry allowed the measurement of differentially-stained cells, and Louis Kamentsky developed the first multiparameter flow cytometer in 1965 to identify and count the cancer cells in cervical tissue specimens.
In...

Controlled-Current Coulometry: Coulometric Titration

Controlled-Current Coulometry: Coulometric Titration

Coulometric titrations are a form of titrimetric analysis where the reagent is generated electrically, and its amount is evaluated based on current and generating time. The electron serves as the standard reagent. The procedure is similar to conventional titrations, such as endpoint detection.
The fundamental requirements for coulometric titrations are (1) 100% efficiency in the reagent-generating electrode reaction and (2) a stoichiometric and preferably rapid reaction between the generated...