High-Throughput Cell Death Assays with Single-Cell and Population-Level Analyses Using Real-Time Kinetic Labeling (SPARKL)

Jesse D Gelles; Jerry E Chipuk

doi:10.1016/j.xpro.2020.100034

High-Throughput Cell Death Assays with Single-Cell and Population-Level Analyses Using Real-Time Kinetic Labeling (SPARKL)

STAR Protoc. 2020 Jun 19;1(1):100034. doi: 10.1016/j.xpro.2020.100034. Epub 2020 Jun 3.

Authors

Jesse D Gelles^{1

2

3}, Jerry E Chipuk^{1

4

2

5

6

7}

Affiliations

¹ Department of Oncological Sciences, Icahn School of Medicine at Mount Sinai, One Gustave L. Levy Place, New York, NY 10029, USA.
² The Tisch Cancer Institute, Icahn School of Medicine at Mount Sinai, One Gustave L. Levy Place, New York, NY 10029, USA.
³ Technical Contact.
⁴ Department of Dermatology, Icahn School of Medicine at Mount Sinai, One Gustave L. Levy Place, New York, NY 10029, USA.
⁵ The Diabetes, Obesity, and Metabolism Institute, Icahn School of Medicine at Mount Sinai, One Gustave L. Levy Place, New York, NY 10029, USA.
⁶ The Graduate School of Biomedical Sciences, Icahn School of Medicine at Mount Sinai, One Gustave L. Levy Place, New York, NY 10029, USA.
⁷ Lead Contact.

Abstract

High-throughput cytostatic and cell death assays are a critical component of pharmacological screens and mechanism-based interrogations into cellular biology. We developed a method for single-cell and population-level analyses using real-time kinetic labeling (abbreviated "SPARKL") with non-toxic fluorescent probes and high-content live-cell imagers. The protocols herein detail the steps, specifics, and suggested utilization of the SPARKL method within several "label-and-go" zero-handling workflows. For complete details on the use and execution of this protocol, please refer to Gelles et al. (2019).

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

MeSH terms

Cell Death*
Cytological Techniques*
Fluorescent Dyes
Kinetics

Substances

Fluorescent Dyes

Abstract

Publication types

MeSH terms

Substances

Grants and funding