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Updated: Aug 9, 2026
Single-cell Analysis of Immunophenotype and Cytokine Production in Peripheral Whole Blood via Mass Cytometry
Published on: June 26, 2018
Simplified immunophenotypic analysis by laser scanning cytometry
R J Clatch1, J R Foreman, J L Walloch
1Christ Hospital and Medical Center, Oak Lawn, Illinois, USA. RJClatch@pol.net
Insights
Laser scanning cytometry offers a new method for immunophenotypic analysis of blood cancers. This technique enhances diagnostic accuracy for leukemia and lymphoma, especially with small specimens.
Area of Science:
- Hematology
- Immunology
- Cytometry
Background:
- Immunophenotypic analysis aids in diagnosing leukemia and lymphoma.
- Flow cytometry is a common method, but has limitations.
- Laser scanning cytometry (LSC) presents a novel alternative.
Purpose of the Study:
- To develop and validate a new specimen preparation and immunofluorescence method for LSC.
- To leverage LSC's capabilities for enhanced hematologic specimen analysis.
- To overcome limitations of traditional flow cytometry.
Main Methods:
- Developed a novel specimen preparation technique specific to LSC.
- Adapted immunofluorescent reactions for LSC.
- Utilized LSC for immunophenotypic analysis of hematologic specimens.
Main Results:
- Enabled analysis of up to 36 antibodies on specimens as small as 50,000 cells.
- Reduced antibody usage by 85% compared to flow cytometry.
- Allowed correlation of cell morphology with physical and fluorescent measurements.
Conclusions:
- The new LSC method is simple, efficient, and cost-effective.
- It is particularly useful for small specimens like fine needle aspirates.
- LSC provides a valuable tool for diagnosing leukemia and lymphoma.
Abstract:
Immunophenotypic analysis of hematologic specimens is a useful laboratory adjunct to surgical pathology and cytology to confirm or further characterize diagnoses of leukemia or lymphoma. Laser scanning cytometry is a new laboratory technology that has been adapted to perform immunophenotypic analysis of hematologic specimens, with numerous advantages as compared with flow cytometry. In order to make full use of the laser scanning cytometer's capabilities, a new method of specimen preparation and means of performing the immunofluorescent reactions was developed. The technique described in this report, specific only to laser scanning cytometry, enables panels of up to 36 different antibodies to be used on specimens as small as 50,000 total cells. The laboratory methodology is simple, requires 85% less antibody than flow cytometric methods, and allows individual cell cytologic morphology to be correlated with objective physical and fluorescent measurements on a cell-by-cell basis. Other advantages are described in the text. Over the course of nine months in our community hospital, we have used this technique clinically to analyze 172 cases of suspected leukemia or lymphoma. The method has proven remarkably useful, particularly for extremely small specimens such as fine needle aspiration biopsies.
