System Components & Performance Summary
Microscope & Camera
Microscope & Camera Functions
Microscope & Camera Performance
Design Requirements
Overview
BD Biosciences designs and manufactures flow cytometers that are largely used in medical and research institutions around the world. Flow cytometers consist of high power lasers that collide with a stream of cells to extract detailed information. The emissions from the cells’ collision with the laser are sent through the pinholes to be analyzed by a computer. This information can be used to determine cell count and biomarker detection for disease and medical research. The alignment and position of these lasers with the cell stream is critical for accurate results.
Over time and through extensive use, lasers become misaligned which may invalidate research data. To achieve successful alignment, field engineers manually tune the laser orientation until it passes horizontally through the center of the pinhole.
To easily observe the laser positions, field engineers from BD Biosciences have been utilizing a simple compound microscope to enhance the field of view. This previous method presented the major safety risk of a magnified stray laser beam entering the microscope and damaging the user’s eye. The goal of this project was to redesign this microscope and provide a new solution to eliminate any potential risk.
Flow Cytometer Laser Alignment System
Carlos Acuña, Justin Chen, Kevin Chen, Daniel Hwang
Department of Mechanical and Aerospace Engineering at University of California, San Diego
Acknowledgements:
Paul Chen - Systems Product Development Engineer at BD Biosciences | Chantell Kuhlmann - Sr. Manager of Systems Engineering at BD Biosciences | Jerry Tustaniwskyj - Professor at UC San Diego | Patrick McGuire - Teaching Assistant | Dennis Sasaki - R&D Associate Director at BD Biosciences, San Diego | Ian Richardson - Machine Shop Technician | Tom Chalfant - Machine Shop Technician
Figure 10: Software determining the positions of the laser and its alignment
Figure 9: Image without PIS (left). Image with PIS activated (right)
Figure 1: Introduction to flow cytometry and laser alignment
Figure 2: Flow Cytometer Laser Alignment System Prototype
Figure 3: Current Solution (left) and Proposed Solution (right)
Figure 4: AD7013MTL Dino-lite Premier
Figure 5: Lit & Unlit Image samples of FACSCanto (top) and FACSAria (bottom)
Mounting Bracket
Bracket Functions
Bracket Performance
Figure 6: System Mounting Bracket
Figure 7: System mounted to FACSCanto (left), FACSAria (middle), and FACSVerse(right)
Software
Software Functions
Software Performance
Future Improvements
Impact on Society
Conclusion
The Flow Cytometer Laser Alignment System is a safe, portable, precise, and inexpensive device that can be easily used to facilitate the laser alignment process of a variety of BD Biosciences flow cytometer product families. This progressive technology embodies a combination of forward thinking engineering and user-centered design that marks the first steps to the enhancement of current outmoded alignment processes.
Pinhole Illumination System (PIS)
PIS Functions
PIS Performance
Figure 8: Pinhole Illumination System
References:
[1] "Introduction to Flow Cytometry." Introduction to Flow Cytometry. Abcam, n.d. Web. 06 June 2014.
[2] Optics, Edmund. "Understanding Microscopes and Objectives." Edmund Optics. N.p., n.d. Web. 06 June 2014
[3] Bose, Arindam. “How to Detect and Track Red Objects in Live Video in MATLAB” http://arindambose.com/blog/?p=72
[4] Becton Dickenson. "Flow Cytometry." Instruments, Software, and Resources. N.p., n.d. Web. 06 June 2014.
[5] Chen, Paul S. Flow Cytometer Laser Imaging Alignment System. San Diego: BD Biosciences, n.d. PDF.