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High-speed photodamage cell selection using a frequency-doubled argon ion laser
J F Keij1, A C Groenewegen, G B Dubelaar
1Department of Molecular Pathology, MBL-TNO, Rijswijk, The Netherlands.
Cytometry
|March 1, 1995
Summary
A novel flow cytometer uses ultraviolet laser light to selectively destroy unwanted cells, enabling high-speed cell sorting with high purity and yield for research applications.
Area of Science:
- Biotechnology
- Cell Biology
- Optical Engineering
Background:
- Previous attempts to photoinactivate cells using photosensitizers were unsuccessful.
- The photosensitivity of cellular DNA at 257 nm was identified as a viable mechanism for cell inactivation.
Purpose of the Study:
- To develop a flow cytometer capable of high-speed cell sorting by selectively photoinactivating undesired cells.
- To evaluate the efficiency and purity of cell selection using ultraviolet laser-induced photodamage.
Main Methods:
- Development of a photodamage flow cytometer (ZAPPER) utilizing a 257 nm laser.
- Irradiation of cells with a focused 257 nm laser beam to induce photodamage.
- Implementation of two sorting modes: Enrichment and Purge, with varying laser targeting and modulation strategies.
Main Results:
- The ZAPPER system achieved selective photoinactivation of cells at rates exceeding 22,000 events/s.
- Selection purities ranged from 81% to 100%, with cell yields of 95-105% in Enrichment mode.
- Reduced yields (12-52%) in Purge mode were attributed to a slow Electro-Optic Modulator and limitations in laser crystal lifetime.
Conclusions:
- The photodamage flow cytometer demonstrates effective high-speed cell sorting through targeted ultraviolet laser irradiation.
- Optimization of sorting modes and hardware components is necessary to improve yields in specific applications.
- The technology shows promise for selective cell manipulation in biological research.