数字全息显微镜在自动化三维细胞识别中的应用概述

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"Digital holographic microscopy for automated 3D cell identification: an overview (Invited Paper)" 是一篇关于数字全息显微镜技术在自动化三维细胞识别中的应用综述。本文详细介绍了数字全息（DH）显微镜技术，这是一种极具潜力的定量相位对比成像方法。DH显微镜的核心在于它能够获取物体波前的复振幅信息，这进一步提供了物体的厚度分布。这一特性使得该技术在生物医学领域，特别是在细胞研究中具有广泛的应用前景。 DH显微镜的一个显著优势是其数值聚焦能力。通过这种方法，可以获取物体在不同轴向平面的厚度分布，这意味着研究人员能够在无需物理移动样品的情况下，对细胞进行多层成像。这对于理解细胞的三维结构和动态变化至关重要。文章中特别提到了双束DH显微镜的应用，这种技术被用来获取不同的细胞参数，以实现细胞成像和自动化细胞识别。通过精确测量细胞的这些参数，如形状、大小、厚度和内部结构等，可以更准确地识别和区分不同的细胞类型，这对于疾病诊断、药物研发和细胞生物学研究有着重要价值。作者Arun Anand和Bahram Javidi在文中概述了他们在该领域的研究成果，强调了DH显微镜在自动化监测和分析细胞过程中的潜力。这项工作不仅促进了生物医学成像技术的发展，也为未来的细胞分析提供了新的工具和方法。这篇综述揭示了数字全息显微镜在细胞研究中的前沿进展，展示了如何利用这项技术实现高精度、高效率的3D细胞识别，对于提高实验效率和推动生命科学的研究具有重要意义。

COL 12(6), 060012(2014) CHINESE OPTICS LETTERS June 10, 2014

Digital holographic microscopy for automated 3D cell

identiﬁcation: an overview

(Invited Paper)

Arun Anand

and Bahram Javidi

2∗

Applied Physics Department, Faculty of Technology and Engineering,

The M.S. University of Baroda, Vadodara 390001, India

Department of Electrical and Computer Engineering, U-2157, University of Connecticut,

Storrs, CT 06269-2157, USA

∗

Corresponding author: bahram@engr.uconn.edu

Received February 25, 2014; accepted April 4, 2014; posted online May 30, 2014

Digital holographic (DH) microscopy is a promising technique for quantitative phase contrast imaging.

It provides complex amplitude of the object wavefront, which in t urn yields the thickness distribution of

the object. An added advantage of the technique is its ability for numerical focusing, which provides the

thickness distribution of the object at diﬀerent axial planes. In this invited paper, we present an overview

of our reported work on two beam DH microscopyto acquire diﬀerent cell parameters for cell imaging and

automated cell identiﬁcation. Applications to automated monitoring of stem cells without destroying the

cells and automated identiﬁcation of malaria infected red blood cells are discussed.

OCIS codes: 090.1995, 180.6900, 120.5050, 170.1530.

doi: 10.3788/COL201412.060012.

1. Introduction

In imaging o f living cells, resolution as well as contrast

is necessary to study and evaluate the dynamic proce sses

occurring in them. But many of the biological speci-

mens are transparent to visible light. For these objects,

bright ﬁeld microscope provides only low contrast inten-

sity images and a single object plane. Staining of the

sp e cimen can improve the co ntrast, but this may cause

an alteration of the cells. These objects may produce a

change to the phase of the probe beam, that is, the beam

interacting with the object, due to spatially varying op-

tical thickness. Thus, phase contrast imaging techniques

will be better suited for cellevaluation. The phase o f the

probe beam is very sensitive to the optical thickness mis-

match between the cytoplasmic content of the cells and

the surrounding medium. This phase change can be used

to construct high contrast images of the specimen

[1,2]

If acce ss to the phase of the object wavefront is avail-

able, optical thickness proﬁle of the specimen can be

reconstructed, leading to quantitative phase microsc opy

(QPM). Digital holographic (DH) microscopy is such a

technique, which provides three dimensional (3D) opti-

cal thickness proﬁles of transparent specimen

[3−15]

. In

DH microscopy, holograms or interference patterns are

recorded on image sensors and reconstructed using nu-

merical methods by simulating the process of diﬀraction

from these structures

[3,4]

. This reco ns truction provides

the complex amplitude of the object wavefront, which in

turn provides the object phase information and hence the

thickness proﬁle. An added advantage of DH microscopy

is its ability of numerical focusing, which provides a way

by which diﬀerent object planes can be brought to focus

during the reconstructio n process. Comparison o f the

object phase with any other phase distribution becomes

possible in the case of DH microscopy. Thus, recon-

structing the phase distribution with and witho ut the

object and then interferometricaly comparing them, the

distortions due to the aberrations in the optical system

may beremoved and phase information due to the object

alone can be obtained

[11−15]

. Also, using a series of holo-

grams the time evolution of the cell morphology could

be studied

[11−13]

. We are involved in the development of

applications using DH microscopy for qua ntitative imag-

ing of cells. In this invited pape r, we pr e sent an overview

of our rep orted and published research in the area of DH

microscopywith applications in the imaging of cells for

their parameter extractio n, co mparison and automated

identiﬁcation

[8,10,11,14,15]

2. Overview of DH microscope

A sketch of the DH microscope used in some of our

investigations is s hown in Fig. 1 (a)

[11−15]

. It uses He-Ne

laser as the source although a variety of other coherent

or partially cohere nt sources may be used. The beam

from the s ource is split into two. One of the beams

acts as the reference beam. The other beam is allowed

to pass through the object under investigation. The

object is magniﬁed using an a ppropriate microscope ob-

jective (selection of the objective lens depends upon the

required magniﬁcation and resolution). The object un-

der investigation is mounted on a transla tion stage for

easy focusing. The o bject and reference beams ar e made

to interfere at the detector plane or the plane of the

hologram at an angle in oﬀ-axis geometry by employ-

ing Mach-Zehnder interferometer conﬁguration. A CCD

array records the ho lograms, that is, the interference

pattern.The detector is located near the image plane of

the objective le ns . To match the curvatures of the wave-

fronts at the detector plane, an objective of the same

conﬁguration as the one used for magniﬁcation was in-

troduced in the reference arm of the setup. Figure 1(b)

shows the portion inside the dotted rectangle shown in

Fig. 1(a), depicting the magniﬁcatio n and interference

process. The dis tance (d) between the hologram and the

image plane is also shown.

1671-7694/2014/060012(6) 060012-1

 2014 Chinese Optics Letters

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数字全息显微镜在自动化三维细胞识别中的应用概述

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