Protocol to measure apoptosis-associated speck-like protein containing a CARD specks in human cerebrospinal fluid via imaging flow cytometry

Kathryn E Sánchez; Shanya Jiang; Sharina Palencia Desai; Jeffery Thompson; Sasha Hobson; Gary A Rosenberg; Kiran Bhaskar

doi:10.1016/j.xpro.2024.102916

Protocol to measure apoptosis-associated speck-like protein containing a CARD specks in human cerebrospinal fluid via imaging flow cytometry

STAR Protoc. 2024 Mar 15;5(1):102916. doi: 10.1016/j.xpro.2024.102916. Epub 2024 Mar 6.

Authors

Kathryn E Sánchez¹, Shanya Jiang², Sharina Palencia Desai², Jeffery Thompson³, Sasha Hobson³, Gary A Rosenberg⁴, Kiran Bhaskar⁵

Affiliations

¹ Center for Memory and Aging, University of New Mexico, Albuquerque, NM 87131, USA; Department of Pediatrics, University of Chicago, Chicago, IL 60637, USA. Electronic address: [email protected].
² Department of Molecular Genetics and Microbiology, University of New Mexico, Albuquerque, NM 87131, USA.
³ Center for Memory and Aging, University of New Mexico, Albuquerque, NM 87131, USA.
⁴ Center for Memory and Aging, University of New Mexico, Albuquerque, NM 87131, USA; Department of Neurology, University of New Mexico, Albuquerque, NM 87131, USA.
⁵ Department of Molecular Genetics and Microbiology, University of New Mexico, Albuquerque, NM 87131, USA; Department of Neurology, University of New Mexico, Albuquerque, NM 87131, USA. Electronic address: [email protected].

Abstract

Apoptosis-associated speck-like protein containing a c-terminal caspase activation and recruitment domain (ASC) specks are elevated in the cerebrospinal fluid (CSF) of Alzheimer's disease and related dementias (AD/ADRDs) patients. Here, we present a flow cytometry protocol to quantify ASC specks. We describe steps for fluorescently labeling ASC specks using antibody technology, visualizing with imaging flow cytometry, and gating based on physical characteristics. CSF ASC specks levels positively correlate with phosphorylated tau (Thr181) and negatively correlate with amyloid β ratio (42/40), thus serving as a neuroinflammatory biomarker for diagnosing AD/ADRDs. For complete details on the use and execution of this protocol, please refer to Jiang et al.¹.

Keywords: Clinical Protocol; Flow Cytometry; Molecular Biology; Neuroscience.

MeSH terms

Amyloid beta-Peptides* / metabolism
Apoptosis
CARD Signaling Adaptor Proteins* / metabolism
Flow Cytometry / methods
Humans
Inflammasomes / metabolism

Substances

CARD Signaling Adaptor Proteins
Amyloid beta-Peptides
Inflammasomes

Abstract

MeSH terms

Substances

Grants and funding