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Detection and measurement of a single blood cell surface antigen by thermal lens microscopy
H Kimura1, F Nagao, A Kitamura
1Department of Forensic Medicine, Juntendo University School of Medicine, Tokyo, Japan.
Insights
A new thermal lens microscopy method detects human leukocyte antigens on single blood cells. This sensitive technique images and quantifies specific cell surface molecules for advanced diagnostics.
Area of Science:
- Biomedical Engineering
- Analytical Chemistry
- Immunology
Background:
- Accurate detection of cell surface antigens is crucial for diagnosing various diseases.
- Existing methods often require large cell populations or lack single-cell resolution.
- Human leukocyte antigens (HLA) play a vital role in immune response and transplantation.
Purpose of the Study:
- To develop a highly sensitive method for detecting and quantifying antigens on the surface of individual blood cells.
- To utilize thermal lens microscopy for single-cell antigen analysis.
- To visualize the distribution of specific human leukocyte antigens (HLA-A, -B, -C) on leukocytes.
Main Methods:
- Employing thermal lens microscopy, a technique utilizing a laser-induced thermal-lens effect for spectrometry.
- Using colloidal gold nanoparticles coated with antibodies to specifically stain membrane antigens on leukocytes.
- Developing a novel laser microscope designed for measuring convex surface cells.
Main Results:
- Successfully detected and quantified human leukocyte antigens (HLA-A, -B, and -C) on individual lymphocytes.
- Obtained images showing the distribution of HLA-A, -B, and -C antigens on mononuclear leukocytes.
- Demonstrated the capability to analyze localized antigens and cell-surface-associated molecules at the single-cell level.
Conclusions:
- Thermal lens microscopy provides a powerful and sensitive analytical tool for single-cell antigen detection.
- The developed method enables precise identification and quantification of specific antigens on leukocytes.
- This technique holds significant potential for advancing diagnostics and research in immunology and cell biology.
Abstract:
A highly sensitive method for detection of antigens on the surface of a single blood cell using thermal lens microscopy is described. Colloidal gold, coated with antibody, was used to stain membrane antigens of leukocytes. Human leukocyte antigens on the lymphocytes and mononuclear leukocytes were observed by new thermal lens microscopy, which involves spectrometry using a laser-induced thermal-lens effect. Antigens of HLA-A, -B, and -C loci on the lymphocytes were identified and quantitated using a single cell. The image of HLA-A, -B, and -C antigen distribution on a mononuclear leukocyte was obtained. Our laser microscope, newly devised for measuring convex surface cells, is a powerful analytical tool for detecting and quantitating localized antigens in a single cell and/or cell-surface-associated molecules.