Emerging Tools for Single-Cell Analysis

Emerging Tools for Single-Cell Analysis: Advances in Optical Measurement Technologies

Edited by Gary Durack, J. Paul Robinson Copyright © 2000 Wiley-Liss, Inc.

ISBNs: 0-471-31575-3 (Hardback); 0-471-22484-7 (Electronic)

EMERGING TOOLS FOR

SINGLE-CELL ANALYSIS

CYTOMETRIC CELLULAR ANALYSIS

Series Editors

J. Paul Robinson	George F. Babcock
Purdue University Cytometry	Department of Surgery
Laboratories	University of Cincinnati College
Purdue University	of Medicine
West Lafayette, Indiana	Cincinnati, Ohio

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J. Paul Robinson and George F. Babcock, Volume Editors

Immunophenotyping

Carlton C. Stewart and Janet K. A. Nicholson, Volume Editors

Emerging Tools for Single Cell Analysis: Advances in Optical

Measurement Technologies

Gary Durack and J. Paul Robinson, Volume Editors

Forthcoming Volume in Series

Cellular Aspects of HIV Infection

Andrea Cossarizza and David Kaplan, Volume Editors

EMERGING TOOLS FOR SINGLE-CELL ANALYSIS

ADVANCES IN OPTICAL MEASUREMENT TECHNOLOGIES

Edited by

GARY DURACK

University of Illinois Biotechnology Center

University of Illinois at Urbana-Champaign

Urbana, Illinois

J. PAUL ROBINSON

Purdue University Cytometry Laboratories

Purdue University

West Lafayette, Indiana

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ISBN 0-471-22484-7

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Contents

1

2

3

4

Preface	vii
Contributors	ix
Cell-Sorting Technology	1
Gary Durack
High-Speed Cell Sorting	21
Ger van den Engh
Rare-Event Detection and Sorting of Rare Cells	49
James F. Leary
Applications of High-Speed Sorting for CD34+ Hematopoietic
Stem Cells	73
Thomas Leemhuis and David Adams

5 Microfabricated Fluidic Devices for Single-Cell Handling
and Analysis	95
David J. Beebe

6

7

Single DNA Fragment Detection by Flow Cytometry	115
Robert C. Habbersett, James H. Jett, and Richard A. Keller
Fluorescence Lifetime Imaging: New Microscopy
Technologies	139
Weiming Yu, William W. Mantulin, and Enrico Gratton

8

9

Fluorescence Lifetime Flow Cytometry	175
John A. Steinkamp
Application of Fluorescence Lifetime and Two-Photon
Fluorescence Cytometry	197
Donald J. Weaver, Jr., Gary Durack, Edward W. Voss, Jr.,
and Anu Cherukuri

v

vi

Contents

10

11

12

13

14

15

Probing Deep-Tissue Structures by Two-Photon
Fluorescence Microscopy	221
Chen-Yuan Dong, Ki Hean Kim, Christof Buehler, Lily Hsu,
Hyun Kim, Peter T. C. So, Barry R. Masters, Enrico Gratton, and
Irene E. Kochevar
Limits of Confocal Imaging	239
James B. Pawley
Scanning Near-Field Optical Imaging and Spectroscopy
in Cell Biology	271
Vinod Subramaniam, Achim K. Kirsch, Attila Jenei, and
Thomas M. Jovin
White-Light Scanning Digital Microscopy	291
J. Paul Robinson and Ben Gravely
Illumination Sources	307
Howard M. Shapiro
Camera Technologies for Cytometry Applications	323
Kenneth Castleman
Index	339

Preface

This book owes its existence to the recent resurgence of interest in high-resolution evaluation of single-cell properties. One of our goals in assembling this volume was to re-evaluate where current technology stands at the beginning of a new millennium. Further, we felt there was a need for critical discussion of some of these technologies. Take cell sorting, for example. Flow cytometers have been sorting effectively for over 30 years, ever since Fulwyler implemented electrostatic cell sorting in 1965 from inkjet printing technologies developed by Richard Sweet. Sorting in itself, however, did not originate in the sixties—it had been proposed 30 years previously by Moldavin and attempted by several others more or less successfully as technologies matured. In recent years significant gains have been realized. High-speed sorters, once considered useful only for chromosome separation, have supplanted “traditional” sorters in many environments. Now they are used for the isolation of stem cells, mammalian sperm, and a variety of other materials. We have attempted to take a fairly rational approach to the phenomenon of sorting by bringing together in one volume the experts who have played leading roles in the recent revolution. But this was not the only driving force. We wanted a more in-depth discussion of the analytical component of sorting and other technologies. Although biologists frequently veer away from anything containing an equation, we are dependent upon the verification of the mathematical concepts in the implementation process. Thus, this volume attempts to create a domain that successfully integrates engineering and biology.

Now more then ever, there is a blurring of the interface between technologies. Biomedical engineering, and especially one of its component parts, tissue engineering, is the growth business of engineering today. Miniaturization of electronic components has forever changed the nature of computing. Similarly, microand now nanomachines capable of performing mechanical action are a reality. Future developments in these areas are sure to impact cytometry in many ways. The success of such technologies will be in a systems approach—combining a good understanding of fluorescence detection with high-speed electronics and miniaturization together with a fundamental grasp of the biology.

Where imaging fits into this picture is anyone’s guess, but there is little doubt that imaging is a rapidly advancing component of cellular and molecular biology that has reached previously unattainable heights. New technologies, transformed into usable laboratory instruments, have altered cell biology almost beyond recognition in the last decade of the 20th century. Confocal microscopy, developed nearly 50 years ago

vii

viii

Preface

but only recently commercialized, gave biologists a unique tool for investigating the inner workings of cell systems. More recently, multiphoton microscopy has taken us a few steps further. The ability to continuously monitor cellular activity in a living system using multiphoton techniques creates a paradigm shift for developmental biologists who can now monitor, in real time, the differentiation processes in embryonic development. Combining these technologies with GFP and other cell-tracking molecules has begun yet another revolution in imaging applications.

An interesting theme that has emerged as this book was being put together is that many of the technologies discussed—sorting, confocal microscopy, color scanning microscopy—all had their roots established well over 50 years ago. Only in recent years have they had a discernable impact, presumably because of parallel technological developments and the more recent commercial commitment by companies that have created off-the-shelf systems usable by scientists who then do not necessarily need to be capable of building such systems themselves.

Our final goal was to compare and contrast totally different technologies that work toward similar goals: evaluating the properties of single cells using optically based measurement systems. Within the biological discovery process, the synergy between cytometry, chemistry, and imaging systems is remarkable. Traditional flow cytometry technologies combined with new imaging and detection technologies have exploded into a plethora of combination systems. It is our hope that this book creates a linkage between the engineering and development of the technologies and the fusion of these technologies into exciting and emerging tools for single cell analysis.

Gary Durack, University of Illinois Biotechnology Center

J. Paul Robinson, Purdue University Cytometry Laboratories

Contributors

David Adams

SyStemix, Inc.

Palo Alto, California

David J. Beebe

University of Wisconsin

Madison, Wisconsin

Christof Buehler

Department of Mechanical Engineering

Massachusetts Institute of Technology

Cambridge, Massachusetts

Kenneth Castleman

Perceptive Scientific Instruments, Inc.

League City, Texas

Anu Cherukuri

Department of Biochemistry

Northwestern University

Evanston, Illinois

Chen-Yuan Dong

Department of Mechanical Engineering

Massachusetts Institute of Technology

Cambridge, Massachusetts

Gary Durack

Biotechnology Center

University of Illinois at Urbana-

Champaign

Urbana, Illinois

Enrico Gratton

Laboratory for Fluorescence Dynamics University of Illinois at Urbana-

Champaign

Urbana, Illinois

Ben Gravely

Cosmic Technologies Corporation

Raleigh, North Carolina

Robert C. Habbersett

Cytometry Group

Los Alamos National Laboratory

Los Alamos, New Mexico

Lily Hsu

Department of Mechanical Engineering

Massachusetts Institute of Technology

Cambridge, Massachusetts

Attila Jenei

Department of Molecular Biology Max Planck Institute for Biophysical

Chemistry Goettingen, Germany

James H. Jett

Cytometry Group

Los Alamos National Laboratory

Los Alamos, New Mexico

Thomas M. Jovin

Department of Molecular Biology Max Planck Institute for Biophysical

Chemistry Goettingen, Germany

Richard A. Keller

Cytometry Group

Los Alamos National Laboratory

Los Alamos, New Mexico

ix

	x	Contributors
	Hyun Kim	Howard M. Shapiro
	Department of Mechanical Engineering	Howard M. Shapiro, M.D., P.C.
	Massachusetts Institute of Technology	West Newton, Massachusetts
	Cambridge, Massachusetts
	Ki Hean Kim	Peter T. C. So
	Department of Mechanical Engineering	Department of Mechanical Engineering
	Massachusetts Institute of Technology	Massachusetts Institute of Technology
	Cambridge, Massachusetts	Cambridge, Massachusetts
	Achim K. Kirsch
	Department of Molecular Biology	John A. Steinkamp
	Max Planck Institute for Biophysical	Los Alamos National Laboratories
	Chemistry	Los Alamos, New Mexico
	Goettingen, Germany
	Irene E. Kochevar	Vinod Subramaniam
	Wellman Laboratories of Photomedicine	Department of Molecular Biology
	Massachusetts General Hospital	Max Planck Institute for Biophysical
	Boston, Massachusetts	Chemistry
	James F. Leary	Goettingen, Germany
	University of Texas Medical Branch
	Galveston, Texas	Ger van den Engh
	Thomas Leemhuis	University of Washington
		Seattle, Washington
	Stanford University Medical Center
	Stanford, California
	William W. Mantulin	Edward W. Voss, Jr.
		Department of Microbiology
	Laboratory for Fluorescence Dynamics
		University of Illinois at Urbana-
	University of Illinois at Urbana-
		Champaign
	Champaign
		Urbana, Illinois
	Urbana, Illinois
	Barry R. Masters	Donald J. Weaver, Jr.
	University of Bern
		Department of Microbiology and
	Bern, Switzerland
		Immunology
	James B. Pawley	University of North Carolina-
	Department of Zoology	Chapel Hill
	University of Wisconsin	Chapel Hill, North Carolina
	Madison, Wisconsin
	J. Paul Robinson	Weiming Yu
	Purdue University Cytometry	Laboratory for Fluorescence Dynamics
	Laboratories	University of Illinois at Urbana-
	Purdue University	Champaign
	West Lafayette, Indiana	Urbana, Illinois