Improved cytometric analysis of untouched lung leukocytes by enzymatic liquefaction of sputum samples.

ABSTRACT:

Background

Phenotyping sputum-resident leukocytes and evaluating their functional status are essential analyses for exploring the cellular basis of pathological processes in the lungs, and flow cytometry is widely recognized as the gold-standard technique to address them. However, sputum-resident leukocytes are found in respiratory samples which need to be liquefied prior to cytometric analysis. Traditional liquefying procedures involve the use of a reducing agent such as dithiothreitol (DTT) in temperature-controlled conditions, which does not homogenize respiratory samples efficiently and impairs cell viability and functionality.

Methods

Here we propose an enzymatic method that rapidly liquefies samples by means of generating O₂ bubbles with endogenous catalase.

SUBMITTER: Santopolo G 

PROVIDER: S-EPMC9673301 | biostudies-literature | 2022 Nov

REPOSITORIES: biostudies-literature