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Related Concept Videos

Flow Cytometry01:23

Flow Cytometry

14.5K
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...
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Sputum Studies I: Gram Stain, cytology, and Acid-fast smear and culture01:26

Sputum Studies I: Gram Stain, cytology, and Acid-fast smear and culture

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Sputum studies are a critical part of diagnosing and treating numerous respiratory conditions. These studies involve obtaining sputum samples for analysis to identify pathogenic organisms and assess the presence of abnormal cells indicative of malignant conditions. This lesson will delve into three fundamental sputum studies: Gram Stain, Cytology, and Acid-fast Smear and Culture.
Gram Stain
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Sputum Studies II: Culture and Sensitivity01:20

Sputum Studies II: Culture and Sensitivity

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Description
Sputum culture and sensitivity is a medical procedure used to diagnose bacterial infections in the respiratory tract and select the most appropriate antibiotics for treatment. This process involves analyzing sputum samples of thick and opaque secretions produced in the lungs and airways. These samples are collected from patients and then sent to the laboratory for analysis.
The test can identify various pathogens responsible for respiratory infections, including Streptococcus,...
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Related Experiment Video

Updated: Oct 23, 2025

Quality-Controlled Sputum Analysis by Flow Cytometry
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Quality-Controlled Sputum Analysis by Flow Cytometry

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Quality-Controlled Sputum Analysis by Flow Cytometry.

Marcia Grayson1, Shao-Chiang Lai1, Lydia H Bederka1

  • 1bioAffinity Technologies.

Journal of Visualized Experiments : Jove
|August 23, 2021
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Summary

Flow cytometry offers a rapid, high-throughput method for analyzing sputum cellular content, overcoming limitations of traditional cytology. This technique efficiently excludes debris and squamous epithelial cells (SECs) for improved lung disease diagnostics.

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Area of Science:

  • Immunology
  • Pulmonology
  • Biotechnology

Background:

  • Sputum analysis is crucial for lung health assessment but traditional cytology is time-consuming and often yields inadequate samples due to debris and squamous epithelial cells (SECs).
  • Flow cytometry provides a high-throughput alternative for cellular analysis, capable of excluding unwanted cellular components.

Purpose of the Study:

  • To present an efficient flow cytometry protocol for analyzing sputum cellular content.
  • To establish a method for high-throughput diagnostics of lung diseases using sputum samples.

Main Methods:

  • Developed a protocol to dissociate sputum into single-cell suspensions, followed by antibody staining and fixation.
  • Implemented a gating strategy to exclude debris, dead cells (including SECs), and cell doublets.
  • Characterized hematopoietic and epithelial subsets using CD45 expression and identified lung-specific macrophages for quality control.

Main Results:

  • The protocol successfully processed sputum samples for flow cytometric analysis, enabling exclusion of debris and SECs.
  • Demonstrated the ability to differentiate between hematopoietic and epithelial cell lineages within sputum.
  • Validated the method's applicability across different flow cytometers (Navios EX, LSR II, Lyric) and its adaptability for additional cellular markers.

Conclusions:

  • Flow cytometry offers a robust, efficient, and adaptable method for comprehensive sputum analysis.
  • This protocol enhances the utility of sputum for high-throughput diagnostics of lung diseases.
  • The method provides a reliable quality control measure by identifying lung-specific cells.