Digital Data Acquisition and Processing

Mark A. Naivar1, David W. Galbraith2

1 DarklingX, LLC, Flow Cytometry, Los Alamos, New Mexico, 2 BIO5 Institute and School of Plant Sciences, The University of Arizona, Tucson, Arizona
Publication Name:  Current Protocols in Cytometry
Unit Number:  Unit 10.19
DOI:  10.1002/0471142956.cy1019s71
Online Posting Date:  January, 2015
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Abstract

A flow cytometer is made up of many different subsystems that work together to measure the optical properties of individual cells within a sample. The data acquisition system (also called the data system) is one of these subsystems, and it is responsible for converting the electrical signals from the optical detectors into list‐mode data. This unit describes the inner workings of the data system, and provides insight into how the instrument functions as a whole. Some of the information provided in this unit is applicable to everyday use of these instruments, and, at minimum, should make it easier for the reader to assemble a specific data system. With the considerable advancement of electronics technology, it becomes possible to build an entirely functional data system using inexpensive hobbyist‐level electronics. This unit covers both analog and digital data systems, but the primary focus is on the more prevalent digital data systems of modern flow cytometric instrumentation. © 2015 by John Wiley & Sons, Inc.

Keywords: data system; electronics; pulse; processing; signal; ADC; DSP

     
 
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Table of Contents

  • Introduction
  • General Description
  • Front‐End Electronics
  • Event Detection
  • Event Processing
  • System Design
  • Advanced Digital Signal Processing
  • Summary
  • Literature Cited
  • Figures
     
 
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Materials

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Figures

Videos

Literature Cited

Literature Cited
  Galbraith, D.W. 2009. Simultaneous flow cytometric quantification of plant nuclear DNA contents over the full range of described angiosperm 2C values. Cytometry A 75:692‐698.
  Godavarti, M., Rodriguez, J.J., Yopp, T.A., Lambert, G.M., and Galbraith, D.W. 1996. Automated particle classification based on digital acquisition and analysis of flow cytometric pulse waveforms. Cytometry 24:330‐339.
  Hausmann, M., Crone, M., and Cremer, C. 1996. Depth of field and improved resolution of slit‐scan flow systems. SPIE 2926:297‐307.
  Hiebert, R.D., Jett, J.H., and Salzman, G.C. 1981. Modular electronics for flow cytometry and sorting: The LACEL system. Cytometry 1:337‐341.
  Hoffman, R.A. 2009. Pulse width for particle sizing. Curr. Protoc. Cytom. 50:1.23.1‐1.23.17.
  Houston, J.P., Naivar, M.A., Jenkins, P., and Freyer, J.P. 2012. Capture of fluorescence decay times by flow cytometry. Curr. Protoc. Cytom. 59:1.25.1‐1.25.21.
  Keij, J.F., Rotterdam, A.V., Groenewegen, A.C., Stokdijk, W., and Visser, J.W.M. 1991. Coincidence in high‐speed flow cytometry: Models and measurements. Cytometry 12:398‐404.
  McCoy, J.P., Chambers, W.H., Lakomy, R., Campbell, J.A., and Stewart, C.C. 1991. Sorting minor subpopulations of cells: Use of fluorescence as the triggering signal. Cytometry 12:268‐274.
  Mullikin, J., Norgren, R., Lucas, J., and Gray, J. 1988. Fringe‐scan flow cytometry. Cytometry 9:111‐120.
  Naivar, M.A., Wilder, M.E., Habbersett, R.C., Woods, T.A., Sebba, D.S., Nolan, J.P., and Graves, S.W. 2009. Development of small and inexpensive digital data acquisition systems using a microcontroller‐based approach. Cytometry A 75:979‐989.
  Noguchi, Y., Kashima, S., and Aikata, T. 1993. Beam‐shaping optics for a slit‐scan flow cytometer. Cytometry 14:819‐825.
  Rehse, M.A., Corpuz, S., Heimfeld, S., Minie, M., and Yachimiak, D. 1995. Use of fluorescence threshold triggering and high‐speed flow cytometry for rare event detection. Cytometry 22:317‐322.
  Robinson, R.D., Wheeless, D.M., Hespelt, S.J., and Wheeless, L.L. 1990. System for acquisition and real‐time processing of multidimensional slit‐scan flow cytometric data. Cytometry 11:379‐385.
  Shapiro, H.M., Perlmutter, N.G., and Stein, P.G. 1998. A flow cytometer designed for fluorescence calibration. Cytometry 33:280‐287.
  Snow, C. 2004. Flow cytometer electronics. Cytometry A 57:63‐69.
  van den Engh, G. and Stokdijk, W. 1989. Parallel processing data acquisition‐system for multilaser flow cytometry and cell sorting. Cytometry 10:282‐293.
  Stewart, C.C. and Stewart, S.J. 2003. A software method for color compensation. Curr. Protoc. Cytom. 23:10.15.1‐10.15.12.
  van Oven, C. and Aten, J.A. 1990. Instrument for real‐time pulse‐shape analysis of slit‐scan flow cytometry signals. Cytometry 11:630‐635.
  Wood, J.C.S. 2009. Establishing and maintaining system linearity. Curr. Protoc. Cytom. 47:1.4.1‐1.4.14.
  Zilmer, N.A., Rodriguez, J.J., Yopp, T.A., Lambert, G.M., and Galbraith, D.W. 1995. Flow cytometric analysis using digital signal processing. Cytometry 20:102‐117.
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