RAMAN TWEEZERS-An Overview - INFLIBNETshodhganga.inflibnet.ac.in/bitstream/10603/75343/13/13... · 2018-07-08 · RAMAN TWEEZERS-An Overview ... (EVs) of Dictyostelium discoideum,

CHAPTER 3

RAMAN TWEEZERS-An Overview

Raman tweezers setup is a hybrid system which exploits the nonphysical

trapping capabilities of optical tweezers and analytical advantages of Raman

spectroscopy to study the biochemical changes in soft matter to single cell level

while keeping the cells under a physiological environment to mimics the in vivo

conditions. Raman tweezers is a technique capable of gathering Raman

fingerprints from live cells to facilitate identification, characterization and sorting

of cells/microorganisms and more importantly it can perform disease diagnosis to

single cell level. This chapter details the works done in various fields using Raman

tweezers as a tool and the advancements in the technique that project its potential.

D E S I G N A N D D E V E L O P M E N T O F O P T I C A L T W E E Z E R S I N

C O M B I N A T I O N W I T H R A M A N S P E C T R O S C O P Y

Chapter 3: Raman Tweezers - An Overview

Page 66

3.1. INTRODUCTION

Even though tissue Raman spectroscopy has provided a good understanding about

the biochemistry of certain disorders [1-8], better information can be gathered from the

studies performed on cellular and sub-cellular level. It is not necessary that the tissue

under study has the cells of one type and/or in the same stage of growth. Depending on the

cell type and the growth phase they are in, the biochemistry of cells varies. When one

records the Raman spectra from a tissue, the derived signal is a combination of signals

from different cells illuminated by the laser focal spot. This result into the lowered spatial

resolution and such measurements will give limited information. This limitation has been

overcome by micro-Raman spectroscopy which uses microscope objectives to target

isolated single cells with a laser beam focused to sub-cellular dimensions. But, the

technique requires immobilization of cells which brings them in direct contact with

surface of the container. The physical or chemical methods of immobilization are

commonly followed in micro-Raman measurements. Even though, the cells are maintained

in appropriate media, micro-Raman spectroscopy measurements confirm that the cells are

settled at the bottom of the container. However, in many cases the physical stress derived

from the adhesion of cells to the surface or the chemical stress because of the change in

chemical environment around the cells may induce undesirable, non-specific perturbations

in the structure of biomolecules inside the living cells. Since, the cells are of micron size,

even slight change in the surrounding environment makes drastic changes in cell

organelles/cytoplasmic content. If one has to perform Raman spectroscopy measurements

on the cells while retaining their natural environment, then it is crucial to design the

favorable experimental procedures.

Relatively recent development of optical tweezers, an optical way of immobilizing

the cells has made a breakthrough in the single cell studies. Raman tweezers is a technique

which combines the nonphysical trapping capabilities of optical tweezers with Raman

spectroscopy to understand the biochemistry of single cell, sub-cellular organs and

microorganisms levitated in a physiological medium. In other words, Raman tweezers

spectroscopy is a special type of micro-Raman spectroscopy technique where the

biological sample under study is maintained closer to the in vivo conditions during the

course of experiment. Conventional micro-Raman spectroscopy also confines the studies


Page 67

to cellular and/or sub-cellular levels, but limited by a major drawback that the sample

under study is not necessarily in its normal/live state. In conventional technique the

cells/microorganisms are fixed either mechanically or chemically on to the glass surface

which most of the times brings the cells under mechanical/chemical stress. Hence we end

up recording the spectrum of a cell which is away from its physiological environment and

the biochemical information we gather might be far away from reality.

Even though optical tweezers, in its early days of growth, was utilized to perform

Raman spectroscopy of optically trapped single particles/droplets in 1990s [9-14],

Changan Xie and co-workers [15] in 2002, extended the application of Raman tweezers

spectroscopy in biological studies. They exploited the technique to record the Raman

spectra of blood cells and yeast cells optically levitated in saline solution and a yeast

solution respectively. The paper also reports the spectroscopic difference between live and

dead yeast cells; where the cell death was affected by heating. Near-infrared (NIR)

wavelength at 785 nm was used for both trapping and Raman excitation to overcome the

possible photodamage. The calibration and alignment of the laser tweezers Raman

spectroscopy (LTRS) system was performed by recording the Raman spectra of a

polystyrene bead of ~2 µm diameter suspended in water. One month before to Xie’s work

in 2002, K. Ajito and K. Torimitsu [16] reported LTRS studies on synaptosomes, the

nerve-ending particles (about 500–700 nm in diameter) isolated from a neuron of rat brain,

dispersed in the phosphate buffer solution (PBS). The study revealed the content of

synaptosomes through the Raman peaks at 1445 cm-1

and 1657 cm-1

corresponding to CH2

deformation mode in lipids and the amide I mode in proteins respectively.

In 2003 Xie’s group reported the real-time Raman spectroscopic measurements

performed during a heat induced protein denaturation process in yeast cell and bacteria

when the temperature of the surrounding medium is increased [17]. In this study the

protein denaturation was spectroscopically monitored with the temperature dependent

change in intensity of the phenylalanine peak at 1004 cm-1

. The findings in this study were

cross confirmed by recording Raman spectra of native and heat-denatured solutions of

bovine serum albumin and pure phenylalanine. A similar change in the 1004 cm-1

peak is

observed in BSA spectra, whereas, no change is observed in phenylalanine spectra.

Considering the weak nature of Raman scattering, Alexander et al [18] recorded the


Page 68

surface enhanced Raman spectra of single optically trapped bacterial spores. SERS

enhancement was realized by keeping the bacterial spores in vicinity of the SERS

substrate prepared by conjugating the gold colloids (60 nm) on the glass surface.

3.2. RAMAN SPECTROSCOPY OF CELL ORGANELLES

Spectroscopic studies of living cellular organelles provide information on the

biomolecular dynamics during cellular functions. Raman tweezers is a promising tool to

study the biochemistry of functional organelles in a living cell. Moreover the low

micrometer focal spots can target cellular organelles precisely and provide high spatial

resolution. Xie et al presented the Raman spectra of nucleus in an optically trapped pine

cell. H. Tang and co-worker [19] could record the Raman signatures from an intact

mitochondria isolated from heart, kidney and liver tissues of a rat and on comparison, the

mitochondria from 3 tissue types showed variation in their lipid concentrations. They also

examined the differences between the Raman spectra of intact and Ca2+

damaged

mitochondria and found that most of the Raman peaks from phospholipids and proteins

disappeared after 60 minutes of exposure to Ca2+

solution.

In another study, Huang et al [20] recorded the space-resolved Raman spectra of

living S. Pombe (yeast) cells at different stages of cell cycle, with an intention to elucidate

the molecular compositions of organelles, including nuclei, cytoplasm, mitochondria, and

septa. The observed spatial changes in the Raman signature were correlated to the

organelle specific chemical transformation inside the cell during different stages of cell

cycle. In another study, Ojeda et al [21] recorded the Raman spectroscopy signatures of an

optically trapped chromosome and with the help of generalized discriminate analysis

(GDA) of the Raman spectra, they could distinguish between chromosome #1, #2, and #3.

The chromosome numbering was identified by G-banding. Tatischeff et al [22]

characterized the cell-derived extracellular vesicles (EVs) of Dictyostelium discoideum, an

amoeba and human urinary exosomes using Raman tweezers spectroscopy. Cells release

diverse types of membrane vesicles of endosomal and plasma membrane origin called

extracellular vesicles (EVs) [23]. EVs play an important role in intercellular

communication by serving as transport vehicles for cytosolic proteins, lipids, and RNA

between the cells.


Page 69

3.3. RAMAN SPECTROSCOPY FOR CELL GROWTH MONITORING

Raman tweezers has also been applied to study the growth related biochemical

changes inside the cells. The micro-chemical environment inside the cell experiences a

continuous change during the different phases of cell growth. Better understanding of the

biochemical processes involved in cell growth will help cell biologists to design effective

methodologies for controlling and manipulating the cell cycles. Many Raman tweezers

spectroscopy based reports have been published in this regard [24-31]. G. P. Singh et al

[24] recorded the real-time Raman spectra of optically trapped single, live yeast cell in G0

phase of its cell growth and G1 phase of its cell cycle. Raman spectroscopic monitoring

witnessed the changes in relative intensity of peaks corresponding to lipids, proteins and

RNA. This increase/decrease in peak intensities was correlated to the increase/decrease in

the concentration of respective biomolecules in the G0 phase and late G1 phase.

Z. Tao et al [25] made a remarkable study on Rhodotorula glutinis (R. glutinis), a

pigmented yeast species, using Raman tweezers by monitoring the change in intensity of

Raman peaks of carotenoids along with those of nucleic acids and lipids. Rhodotorula

glutinis is a pigmented yeast species which produces large amount of carotenoids and has

industrial applications. The carotenoid accumulation in R. glutinis is known to depend on

the culture conditions and stage of the cell growth. Tao’s data showed that the carotenoid

production is more in the late exponential and stationary phases of cell growth represented

by the increase in lipid concentration with concomitant decrease in DNA/RNA

concentration. Biochemical synthesis in glucose stimulated rat β-cells was monitored

using Raman tweezers spectroscopy by X. Rong and coworkers [26]. The data in this

study suggested a time dependent increase in intensity of Raman peaks corresponding to

proteins and lipids as a representative of the synthesis of those biomolecules in glucose

stimulated cells.

H. Wu et al [27] demonstrated the possibility of using Raman tweezers for real-

time, in vivo lipidomics in oil-producing microalgae, the future source of bio-fuels. He

addressed the problem of extracting the lipids from microalgae to check the production

efficiency for various culture conditions and at different phases of cell growth. Raman

tweezers can replace the time consuming chemical techniques to provide relatively fast


Page 70

monitoring of lipid concentration in microalgae. In an another report, Yan Li and group

[28] studied the growth dynamics for over 40 min and heterogeneity in two interacting

microbial cells using dual trap Raman tweezers setup which can simultaneously record the

spectra from both the cells. Biochemical changes related to the growth dynamics of a

daughter and parent cells in a budding yeast cell have been monitored with time and

heterogeneity in the cell content was presented.

Heterogeneity in the release of CaDPA during the germination of two Bacillus

spores derived from identical microenvironment was also monitored. P. Zhang et al [29]

also studied the dynamics and heterogeneity during the germination of individual bacterial

spores using Raman tweezers and quantitative differential interference contrast

microscopy (DICM). Another study lead by L. Kong et al [30] combined phase contrast

microscopy with Raman tweezers to monitor a variety of changes that occurred during the

germination of single Bacillus spores in both nutrient (l-alanine) and non-nutrient (Ca-

dipicolinic acid) germinants with a temporal resolution of 2s. A study by Zhou et al [31],

combining Raman tweezers and DICM showed that the kinetics of germination in

individual G. stearothermophilus spores are generally similar to that of Bacillus species.

The study could find Raman markers for germination of G. Stearothermophilus spores at

different activation temperatures. A recent study by same group extended the use of

Raman tweezers and DICM to compare the kinetics of germination in Clostridium difficile

spores with that in Bacillus subtilis [32]. Clostridium difficile is a leading cause of

nosocomial diarrhea and germination of Clostridium difficile spores by the bile salts of

gastrointestinal tract initiate the infection.

3.4. SPECTROSCOPIC MONITORING OF THE CELL UNDER STRESS

Biological cells experience many types of stress induced by the extracellular and

intracellular environment [33]. Some aspects which can induce stress in cells include the

hyperosmotic pressure [34, 35], chemical stress because of reactive oxygen species [33],

mechanical stress because of cell packing and cell movement in constricted blood veins

[36]; stress induced by temperature and pH changes [37, 38] etc. Under these stress

conditions, cells will experience drastic changes in structures and conformations of

various intracellular biomolecules. These biochemical changes can represent either the


Page 71

surrender of cells to the stress or cells’ protective response to the stress for their survival.

Careful monitoring of the biochemical changes can be helpful to address stress related

disorders. Application of Raman tweezers left no biological field untouched because of its

high specificity and versatility [39]. Again Raman tweezers wins the race over other

techniques to monitor cells’ stress response because it can extract in situ biochemical

fingerprint from a single cell.

G.P Singh et al [40] have performed Raman tweezers study on hyperosmotic stress

condition by taking yeast cell as a model. In this report they induced hyperosmotic stress

on yeast cells by diluting them in 10% glucose solution made up in synthetic defined

complete (SDC) media. They observed an enhanced production of glycerol and ethanol in

the yeast cells that are subjected to hyperosmotic stress conditions. The effect of

hyperosmotic alcohol solution on single human red blood cells (RBCs) was studied by J.

L. Deng et al [41]. Real-time monitoring of Raman fingerprints from a trapped RBC in

20% alcohol solution showed a time dependent decrease in intensity of 752 cm-1

peak

arriving from porphyrin breathing mode. The rate of decrease further increased with

increase in concentration of alcohol solution and RBCs got ruptured (lysed) after being 3

minutes in 30% alcohol solution.

Raman tweezers, mostly in its dual or multiple trap mode has been successively

employed to study the in situ biochemical response of a cell to the mechanically induced

stress [42-46]. Most of these studies used erythrocytes/red blood cells (RBCs) as model

cell, because the mechanical rigidity of RBCs leads to the oxygen transport related

complications in many diseases including malaria, spherocytosis, elliptocytosis, sickle cell

anemia etc. Erythrocytes adopt different squeezing/stretching states during their passage

through tiny blood capillaries to deliver oxygen and hence the study of mechanochemical

processes associated with deformation of RBCs is necessary. The results of two

independent studies conducted by Satish Rao et al [43] and G. Rusciano [44] showed that

the mechanical stretching of RBCs induces hemoglobin transition from oxygenated state

to the deoxygenated state. Whereas, the Raman tweezers experiments performed by S. Raj

et al [45, 46] on the RBCs under varying percentage of mechanical stretching showed a

significant, load dependent, increase in the intensity of 1035 cm-1

peak corresponding to

in-plane CH2 asymmetric deformation and/or phenylalanine along with changes in some


Page 72

other peaks. These studies postulated the possible role of membrane proteins in controlling

the RBC shape change by releasing the spectrin protein [45, 46].

Many biological systems experience oxidative stress and have their own

antioxidative system to suppress its adverse effects. Oxidative stress pathway is through

the production of intermediate reaction species like peroxides, free radicals etc collectively

called reactive oxygen species (ROS) which damage the structure and properties of cell

components [47, 48]. Either the elevated level of these species inside the cell or failure of

its antioxidant system leads to the permanent cell damage. Apart from their negative role,

reactive oxygen species play a pivotal role in immune system to attack and kill the

pathogens [49] . There are techniques to probe the effect of oxidative stress on cells and/or

the protective response of cells to it, but most of these techniques will be employed on a

large population of cells and lack in situ measurements. Raman tweezers can focus its

attention on single, live cell and has capabilities to depict the stress dependent biochemical

reactions [48, 50].

3.5. IDENTIFICATION, CHARACTERISATION AND SORTING OF

MICROBES

The advancements in socially vital fields like clinical diagnosis, pharmaceutical

industry, food and water processing industries etc demand a rapid tool for detection,

identification and characterization of both good and bad microorganisms. In this aspect

Raman tweezers will play a crucial role as it can give very specific and in situ information

about different species of microorganisms in comparatively less time. Rapid microbial

identification is more necessary in medicine for fast diagnosis and early medical

intervention to various infections. Researchers have already initialized Raman tweezers

based fundamental studies in this direction [51-56]. The studies conducted by C. Xie and

coworkers [52] demonstrated the identification of optically trapped bacterial spores based

on their Raman signatures and also achieved the optical sorting of the spores by optically

manipulating them through the micro-channel into a clean collection chamber. In another

study, W. E. Huang et al [55] identified and sorted two strains of bacteria (P. Fluorescens

and E.coli ) on the basis of 13

C uptake affected red shift of phenylalanine peak from 1001

cm-1

to 965 cm-1

in P. Fluorescens Raman spectra. To demonstrate the specificity of


Page 73

Raman tweezers, optically separated bacteria were later utilized for single strain

cultivation and single cell genome amplification. The study implies that the Raman

tweezers approach has potential application in picking and studying the nonculturable

microorganisms that make 99% of all microbes present in the environment. Recently, Pilát

et al [57] added their contribution to this field by developing and demonstrating an

analytical sorting system combining optical trapping and Raman spectroscopy which

works in micro-fluidic environment to identify and sort living cells of various prokaryotic

and eukaryotic origins. A report by Huang et al [58] have projected the potential

applications of Raman-activated cell sorting (RACS) in single cell biotechnology with

emphases on development of bench top, integrated Raman tweezers systems capable of

extracting DNA/RNA from single cell for sequencing.

The gene expression in living organisms is accompanied by a lot of biochemical

changes in intracellular environment. Raman tweezers spectroscopy was applied to

monitor the dynamics of gene expression and protein synthesis in live E. Coli bacterial

cells by triggering the over expression of Myelin oligodendrocyte glycoprotein [MOG (1-

120)] on exposure to isopropyl thiogalactoside (IPTG) [59]. The study observed a time-

dependent increase in the intensity of characteristic Raman peaks of vibrations in proteins.

The in vivo, real-time uptake and metabolism of trehalose, a disaccharide having two α-

glucose units by Sinorhizobium meliloti bacteria is quantified by holding the live

bacterium in place with an optical trap and recording the Raman spectra at various time

intervals after supplementing the medium with trehalose [60]. In general, this approach

gives real-time chemical information and nullifies the toxic effects of isotopic probes

involved in cellular uptake studies.

On the other hand, Raman tweezers have been used to determine the spectral

changes associated with biochemical response of bacteria to the antibiotics [61, 62]. In a

recent study by Bernatová et al [63, 64], the influence of selected bacteriostatic and

bactericidal antibiotic agents on Staphylococcus epidermidis bacteria is monitored using

Raman tweezers. They observed little change in the Raman signals of DNA in the spectra

of Staphylococcus epidermidis treated with a bacteriostatic agent whereas the action of

bactericidal agent decreased the signal strength from DNA drastically; suggesting DNA

fragmentation as one of the pathways of bactericidal action. The study also extended to


Page 74

demonstrate the ability of Raman tweezers to distinguish between biofilm-positive and

biofilm-negative strains of Staphylococcus epidermidis with the help of principal

component analysis (PCA) [65, 66].

3.6. IDENTIFICATION AND CHARACTERISATION OF DISEASES

Single cell Raman spectroscopy studies will provide better insight into biochemical

modifications due to various diseases and thereby provide adequate, useful resources for

drug design. Even the action of drugs to repair the disease induced biochemical damage

can be well understood from the Raman spectroscopy studies on drug treated cells

maintained under in vivo conditions. Lots of attempts have been made to identify and

characterize the Raman shift markers for various diseases including cancer and mode of

action of drugs on these diseases. In some early attempts, K. E. Hamden et al applied

Raman tweezers to characterize Kaposi sarcoma (KS), a tumor caused by human

herpesvirus-8 (HHV8) [67] and the study revealed a significant variations in intensity of

some peaks between the Raman spectra of normal and infected cells. In another interesting

study on Kaposi’s sarcoma, the Raman fingerprint of cells supporting Kaposi’s sarcoma-

associated herpesvirus (KSHV) reactivation has been reported by recording the Raman

spectra from trapped normal and KSHV infected cells [68].

Studies carried out by K. Chen et al [69] and F. Zheng et al [70] applied the

technique for diagnosis of colorectal cancer. The former study used PCA as a

discrimination tool, whereas the later study used artificial neural networking for the

classification of spectral features from cancerous and noncancerous cells. J. W. Chan and

co-workers have reported the studies on Raman tweezers spectroscopy of normal and

leukemic cells [71-73] . In their first study [71] they compared the Raman spectral features

of leukemic cells derived from transformed Raji (B) and Jurkat (T) cell lines with those of

normal cells derived from blood of healthy volunteers. The second report from Chan et al

[72] reported the Raman spectroscopic analysis of the hematopoietic cells obtained from

four healthy individuals and three leukemia patients. The third study focuses on the

advantages of Raman spectroscopy of optically trapped cells over that of chemically fixed

cells and the possible anomalies in the spectroscopic discrimination of para-formaldehyde

and methanol fixed normal and leukemia cells. Raman spectroscopic identification of


Page 75

different healthy and unhealthy single cells in a suspension prepared using peripheral

blood of leukemia patients have been achieved [74]. A. Y. Lau et al [75] integrated

multichannel micro-fluidics with Raman tweezers for identification, and simultaneous

sorting of individual cells from two leukemia cell lines based on their Raman shift

markers. Raman Tweezers literature also quotes the identification and characterization of

carcinoma of prostate [76, 77], bladder [76], brain [78], pharynx [79], and liver [80].

Probing the response of cancer cells to various treatments including chemotherapy,

radiotherapy and drugs is also necessary. These studies provide the information which

plays a vital role in deciding the dose, efficiency of the treatment, post treatment effects

and counter actions. The induced apoptosis in gastric carcinoma cells by 5-FU (5-

fluorouracil), a drug that is known to induce the apoptosis of cancer cells has been

analyzed using Raman tweezers [81]. The observations of the study suggested a significant

decrease in the Raman peak intensity of cellular lipids, proteins and nucleic acids owing to

the cell death. Tobias J. Moritz et al [82] performed the Raman spectroscopic analyses of

leukemic T cells exposed to the chemotherapy drug doxorubicin at different time points

over 72 hours. They reported the increase in intensities of lipid and DNA Raman peaks

with drug exposure time and concentration. A recent study in this category is reported by

Liu et al [83], which focuses on Raman spectroscopic probing of oxygenation response of

optically trapped single, RBCs derived from normal adult, sickle cell anaemia, and cord

blood to an applied mechanical force. A force dependent increase in the deoxygenation is

observed in all the three cell types; with sickle RBCs showing more deoxygenation,

normal RBCs showing less deoxygenation and cord blood RBCs showing least

deoxygenation for the same optical forces.

3.7. ADVANCES IN RAMAN TWEEZERS

Researchers are in verge of developing many complimentary techniques to

increase the capabilities of Raman tweezers. One of such approaches is the development

of computer algorithms to control the positioning of stage and acquisition of Raman

spectra in holographic Raman tweezers by hand tracking, gestures recognition, eye

tracking and speech recognition [84, 85]. The other report is on developing a lab-on-chip

optical trapping and Raman spectroscopy setup using micro-fabricated dual waveguides


Page 76

[86]. In this chip, a loop shaped waveguide in which the trapping and Raman excitation

laser is coupled bisects a microfluidic channel where both the waveguide and channel are

fabricated monolithically on a SiO2 substrate. The light coming from two arms of the

waveguide traps and Raman excites the micro-particles flowing through the channel. The

scattered Raman signals in the transverse direction were collected using a microscope

objective and fed into a spectrograph. Another study reports the Raman spectroscopic

characterization and identification of micro-particles that are trapped and propelled along

the optical waveguides due to radiation pressure generated by evanescent field that is built

above the waveguide surface [87]. Lihui Ren and co-workers [88] designed an automatic

system for Raman-based single-cell phenotyping and demonstrated its use in

discriminating between Saccharomyces cerevisiae and BY4743 Streptococcus sanguinis

both in tube and slide environment. The automation software framework includes

algorithms for instrument control, image analysis, Raman profiling, database update and

database search. In an interesting study, Wheaton et al [89] have used double-nanohole

optical tweezers to record the extraordinary acoustic Raman (EAR) spectra of trapped

polystyrene nanosphere, titania nanosphere and globular proteins (carbonic anhydrase and

conalbumin). Double-nanohole optical tweezers has also been used to record the Raman

vibrational spectra of a single MS2 bacteriophage [90]. In another report, Yuan and co-

workers have claimed the designing of an integrated microscopy system that has

capabilities to perform Raman tweezers spectroscopy measurements on a single cell along

with the ability to capture the real-time images from trapped cell in both reflectance

confocal microscopy (RCM) mode and two photon fluorescence (TPF) mode [91].


Page 77

3.8. REFERENCES

[1] R. Richards-Kortum and E. Sevick-Muraca, "Quantitative optical spectroscopy for

tissue diagnosis," Annual Review of Physical Chemistry, vol. 47, pp. 555-606,

1996.

[2] N. Stone, C. Kendall, J. Smith, P. Crow, and H. Barr, "Raman spectroscopy for

identification of epithelial cancers," Faraday Discussions, vol. 126, pp. 141-157,

2004.

[3] D. I. Ellis and R. Goodacre, "Metabolic fingerprinting in disease diagnosis:

biomedical applications of infrared and Raman spectroscopy," Analyst, vol. 131,

pp. 875-885, 2006.

[4] Z. Movasaghi, S. Rehman, and I. U. Rehman, "Raman spectroscopy of biological

tissues," Applied Spectroscopy Reviews, vol. 42, pp. 493-541, 2007.

[5] C. Krafft, G. Steiner, C. Beleites, and R. Salzer, "Disease recognition by infrared

and Raman spectroscopy," Journal of Biophotonics, vol. 2, pp. 13-28, 2009.

[6] I. Ur-Rehman, Z. Movasaghi, and S. Rehman, Vibrational spectroscopy for tissue

analysis: CRC Press, 2012.

[7] A. C. S. Talari, Z. Movasaghi, S. Rehman, and I. Ur-Rehman, "Raman

spectroscopy of biological tissues," Applied Spectroscopy Reviews, vol. 50, pp. 46-

111, 2015.

[8] V. B. Kartha and S. Chidangil, Biomedical spectroscopy. Manipal, India: Manipal

University Press, 2014.

[9] G. Schweiger, "Raman scattering on microparticles: size dependence," JOSA B,

vol. 8, pp. 1770-1778, 1991.

[10] M. Lankers, J. Popp, and W. Kiefer, "Raman and fluorescence spectra of single

optically trapped microdroplets in emulsions," Applied Spectroscopy, vol. 48, pp.

1166-1168, 1994.

[11] E. Urlaub, M. Lankers, I. Hartmann, J. Popp, M. Trunk, and W. Kiefer, "Raman

investigation of styrene polymerization in single optically trapped emulsion

particles," Chemical Physics Letters, vol. 231, pp. 511-514, 1994.

[12] M. Lankers, J. Popp, E. Urlaub, H. Stahl, G. Rössling, and W. Kiefer,

"Investigations of multiple component systems by means of optical trapping and

Raman spectroscopy," Journal of Molecular Structure, vol. 348, pp. 265-268,

1995.

[13] C. Esen, T. Kaiser, and G. Schweiger, "Raman investigation of

photopolymerization reactions of single optically levitated microparticles," Applied

Spectroscopy, vol. 50, pp. 823-828, 1996.


Page 78

[14] J. F. Lübben, C. Mund, B. Schrader, and R. Zellner, "Uncertainties in temperature

measurements of optically levitated single aerosol particles by Raman

spectroscopy," Journal of Molecular Structure, vol. 480, pp. 311-316, 1999.

[15] C. Xie, M. A. Dinno, and Y.-q. Li, "Near-infrared Raman spectroscopy of single

optically trapped biological cells," Optics Letters, vol. 27, pp. 249-251, 2002.

[16] K. Ajito and K. Torimitsu, "Laser trapping and Raman spectroscopy of single

cellular organelles in the nanometer range," Lab on a Chip, vol. 2, pp. 11-14, 2002.

[17] C. Xie, Y.-q. Li, W. Tang, and R. J. Newton, "Study of dynamical process of heat

denaturation in optically trapped single microorganisms by near-infrared Raman

spectroscopy," Journal of Applied Physics, vol. 94, pp. 6138-6142, 2003.

[18] T. A. Alexander, P. M. Pellegrino, and J. B. Gillespie, "Near-infrared surface-

enhanced-Raman-scattering-mediated detection of single optically trapped

bacterial spores," Applied Spectroscopy, vol. 57, pp. 1340-1345, 2003.

[19] H. Tang, H. Yao, G. Wang, Y. Wang, Y.-q. Li, and M. Feng, "NIR Raman

spectroscopic investigation of single mitochondria trapped by optical tweezers,"

Optics Express, vol. 15, pp. 12708-12716, 2007.

[20] Y.-S. Huang, T. Karashima, M. Yamamoto, and H.-o. Hamaguchi, "Molecular-

level investigation of the structure, transformation, and bioactivity of single living

fission yeast cells by time-and space-resolved Raman spectroscopy," Biochemistry,

vol. 44, pp. 10009-10019, 2005.

[21] J. F. Ojeda, C. Xie, Y. Q. Li, F. E. Bertrand, J. Wiley, and T. J. McConnell,

"Chromosomal analysis and identification based on optical tweezers and Raman

spectroscopy," Optics Express, vol. 14, pp. 5385-5393, 2006.

[22] I. Tatischeff, E. Larquet, J. M. Falcón-Pérez, P.-Y. Turpin, and S. G. Kruglik, "Fast

characterisation of cell-derived extracellular vesicles by nanoparticles tracking

analysis, cryo-electron microscopy, and Raman tweezers microspectroscopy,"

Journal of Extracellular Vesicles, vol. 1, pp. 19179-19179-11, 2012.

[23] G. Raposo and W. Stoorvogel, "Extracellular vesicles: exosomes, microvesicles,

and friends," The Journal of Cell Biology, vol. 200, pp. 373-383, 2013.

[24] G. P. Singh, G. Volpe, C. M. Creely, H. Grötsch, I. M. Geli, and D. Petrov, "The

lag phase and G1 phase of a single yeast cell monitored by Raman

microspectroscopy," Journal of Raman Spectroscopy, vol. 37, pp. 858-864, 2006.

[25] Z. Tao, G. Wang, X. Xu, Y. Yuan, X. Wang, and Y. Li, "Monitoring and rapid

quantification of total carotenoids in Rhodotorula glutinis cells using laser

tweezers Raman spectroscopy," FEMS Microbiology Letters, vol. 314, pp. 42-48,

2011.


Page 79

[26] X. Rong, S. S. Huang, X. C. Kuang, and H. Liu, "Real‐time detection of

single‐living pancreatic β‐cell by laser tweezers Raman spectroscopy: High

glucose stimulation," Biopolymers, vol. 93, pp. 587-594, 2010.

[27] H. Wu, J. V. Volponi, A. E. Oliver, A. N. Parikh, B. A. Simmons, and S. Singh,

"In vivo lipidomics using single-cell Raman spectroscopy," Proceedings of the

National Academy of Sciences, vol. 108, pp. 3809-3814, 2011.

[28] Y. Li, G. Wang, H.-l. Yao, J. Liu, and Y.-q. Li, "Dual-trap Raman tweezers for

probing dynamics and heterogeneity of interacting microbial cells," Journal of

Biomedical Optics, vol. 15, pp. 067008-067008-7, 2010.

[29] P. Zhang, L. Kong, G. Wang, P. Setlow, and Y.-q. Li, "Combination of Raman

tweezers and quantitative differential interference contrast microscopy for

measurement of dynamics and heterogeneity during the germination of individual

bacterial spores," Journal of Biomedical Optics, vol. 15, pp. 056010-056010-9,

2010.

[30] L. Kong, P. Zhang, P. Setlow, and Y.-q. Li, "Characterization of bacterial spore

germination using integrated phase contrast microscopy, Raman spectroscopy, and

optical tweezers," Analytical Chemistry, vol. 82, pp. 3840-3847, 2010.

[31] T. Zhou, Z. Dong, P. Setlow, and Y.-q. Li, "Kinetics of germination of individual

spores of Geobacillus stearothermophilus as measured by Raman spectroscopy and

differential interference contrast microscopy," PloS One, vol. 8, p. e74987, 2013.

[32] S. Wang, A. Shen, P. Setlow, and Y.-q. Li, "Characterization of the dynamic

germination of individual Clostridium difficile spores using Raman spectroscopy

and differential interference contrast microscopy," Journal of Bacteriology, pp. JB.

00200-15, 2015.

[33] H. Sone and H. Akanuma, "Oxidative stress-mediated signaling pathways by

environmental stressors," in Oxidative stress in vertebrates and invertebrates:

Molecular aspects of cell signaling, T. Farooqui and A. A. Farooqui, Eds., ed:

Wiley-Blackwell, 2011, pp. 175-194.

[34] H. Saito and F. Posas, "Response to hyperosmotic stress," Genetics, vol. 192, pp.

289-318, 2012.

[35] M. B. Burg, J. D. Ferraris, and N. I. Dmitrieva, "Cellular response to hyperosmotic

stresses," Physiological Reviews, vol. 87, pp. 1441-1474, 2007.

[36] S. Suresh, "Mechanical response of human red blood cells in health and disease:

some structure-property-function relationships," Journal of Materials Research,

vol. 21, pp. 1871-1877, 2006.


Page 80

[37] I. Ivanov, "Low pH-induced hemolysis of erythrocytes is related to the entry of the

acid into cytosole and oxidative stress on cellular membranes," Biochimica et

Biophysica Acta (BBA)-Biomembranes, vol. 1415, pp. 349-360, 1999.

[38] J. F. Davidson, B. Whyte, P. H. Bissinger, and R. H. Schiestl, "Oxidative stress is

involved in heat-induced cell death in Saccharomyces cerevisiae," Proceedings of

the National Academy of Sciences, vol. 93, pp. 5116-5121, 1996.

[39] L. Tong, K. Ramser, and M. Käll, "Optical tweezers for Raman spectroscopy," in

Raman spectroscopy for nanomaterials characterization, ed: Springer, 2012, pp.

507-530.

[40] G. P. Singh, C. M. Creely, G. Volpe, H. Grotsch, and D. Petrov, "Real-time

detection of hyperosmotic stress response in optically trapped single yeast cells

using Raman microspectroscopy," Analytical Chemistry, vol. 77, pp. 2564-2568,

2005.

[41] J. L. Deng, Q. Wei, M. H. Zhang, Y. Z. Wang, and Y. Q. Li, "Study of the effect of

alcohol on single human red blood cells using near-infrared laser tweezers Raman

spectroscopy," Journal of Raman Spectroscopy, vol. 36, pp. 257-261, 2005.

[42] D. Cojoc, E. Ferrari, V. Garbin, and E. Di Fabrizio, "Multiple optical tweezers for

micro Raman spectroscopy," in Proceedings of SPIE, 2005, pp. 64-74.

[43] S. Rao, Š. Bálint, B. Cossins, V. Guallar, and D. Petrov, "Raman study of

mechanically induced oxygenation state transition of red blood cells using optical

tweezers," Biophysical Journal, vol. 96, pp. 209-216, 2009.

[44] G. Rusciano, "Experimental analysis of Hb oxy–deoxy transition in single optically

stretched red blood cells," Physica Medica, vol. 26, pp. 233-239, 2010.

[45] S. Raj, M. Marro, M. Wojdyla, and D. Petrov, "Mechanochemistry of single red

blood cells monitored using Raman tweezers," Biomedical Optics Express, vol. 3,

pp. 753-763, 2012.

[46] S. Raj, M. Wojdyla, and D. Petrov, "Studying single red blood cells under a

tunable external force by combining passive microrheology with Raman

spectroscopy," Cell Biochemistry and Biophysics, vol. 65, pp. 347-361, 2013.

[47] V. K. Sharma, "Reactive oxygen species," in Oxidation of amino acids, peptides,

and proteins, ed: John Wiley & Sons, Inc., 2012, pp. 122-204.

[48] E. Zachariah, A. Bankapur, S. Chidangil, M. Valiathan, and D. Mathur, "Probing

oxidative stress in single erythrocytes with Raman Tweezers," Journal of

Photochemistry and Photobiology B: Biology, vol. 100, pp. 113-116, 2010.

[49] C. Nathan and A. Cunningham-Bussel, "Beyond oxidative stress: an

immunologist's guide to reactive oxygen species," Nature Reviews Immunology,

vol. 13, pp. 349-361, 2013.


Page 81

[50] W. T. Chang, H. L. Lin, H. C. Chen, Y. M. Wu, W. J. Chen, Y. T. Lee, et al.,

"Real‐time molecular assessment on oxidative injury of single cells using Raman

spectroscopy," Journal of Raman Spectroscopy, vol. 40, pp. 1194-1199, 2009.

[51] J. Chan, A. Esposito, C. Talley, C. Hollars, S. Lane, and T. Huser, "Reagentless

identification of single bacterial spores in aqueous solution by confocal laser

tweezers Raman spectroscopy," Analytical Chemistry, vol. 76, pp. 599-603, 2004.

[52] C. Xie, D. Chen, and Y.-q. Li, "Raman sorting and identification of single living

micro-organisms with optical tweezers," Optics Letters, vol. 30, pp. 1800-1802,

2005.

[53] C. Xie, J. Mace, M. Dinno, Y. Li, W. Tang, R. Newton, et al., "Identification of

single bacterial cells in aqueous solution using confocal laser tweezers Raman

spectroscopy," Analytical Chemistry, vol. 77, pp. 4390-4397, 2005.

[54] R. Pätzold, M. Keuntje, K. Theophile, J. Müller, E. Mielcarek, A. Ngezahayo, et

al., "In situ mapping of nitrifiers and anammox bacteria in microbial aggregates by

means of confocal resonance Raman microscopy," Journal of Microbiological

Methods, vol. 72, pp. 241-248, 2008.

[55] W. E. Huang, A. D. Ward, and A. S. Whiteley, "Raman tweezers sorting of single

microbial cells," Environmental Microbiology Reports, vol. 1, pp. 44-49, 2009.

[56] P. Zhang, P. Setlow, and Y. Li, "Characterization of single heat-activated Bacillus

spores using laser tweezers Raman spectroscopy," Optics Express, vol. 17, pp.

16480-16491, 2009.

[57] Z. Pilát, J. Ježek, J. Kaňka, and P. Zemánek, "Raman tweezers in microfluidic

systems for analysis and sorting of living cells," in SPIE BiOS, 2014, pp. 89471M-

89471M-9.

[58] W. E. Huang, Y. Song, and J. Xu, "Single cell biotechnology to shed a light on

biological ‘dark matter’in nature," Microbial Biotechnology, vol. 8, pp. 15-16,

2015.

[59] J. W. Chan, H. Winhold, M. H. Corzett, J. M. Ulloa, M. Cosman, R. Balhorn, et

al., "Monitoring dynamic protein expression in living E. coli. bacterial cells by

laser tweezers Raman spectroscopy," Cytometry Part A, vol. 71, pp. 468-474,

2007.

[60] A. Avetisyan, J. B. Jensen, and T. Huser, "Monitoring Trehalose Uptake and

Conversion by Single Bacteria using Laser Tweezers Raman Spectroscopy,"

Analytical Chemistry, vol. 85, pp. 7264-7270, 2013.

[61] T. J. Moritz, D. S. Taylor, C. R. Polage, D. M. Krol, S. M. Lane, and J. W. Chan,

"Effect of cefazolin treatment on the nonresonant Raman signatures of the


Page 82

metabolic state of individual Escherichia coli cells," Analytical Chemistry, vol. 82,

pp. 2703-2710, 2010.

[62] T. J. Moritz, C. R. Polage, D. S. Taylor, D. M. Krol, S. M. Lane, and J. W. Chan,

"Evaluation of Escherichia coli cell response to antibiotic treatment by use of

Raman spectroscopy with laser tweezers," Journal of Clinical Microbiology, vol.

48, pp. 4287-4290, 2010.

[63] S. Bernatova, O. Samek, Z. Pilat, M. Sery, J. Jezek, P. Jakl, et al., "Raman

tweezers on bacteria: following the mechanisms of bacteriostatic versus

bactericidal action," in Proceedings of SPIE, 2014, pp. 91291Y-91291Y-7.

[64] S. Bernatová, O. Samek, Z. Pilát, M. Šerý, J. Ježek, P. Jákl, et al., "Following the

mechanisms of bacteriostatic versus bactericidal action using raman spectroscopy,"

Molecules, vol. 18, pp. 13188-13199, 2013.

[65] O. Samek, P. Zemanek, S. Bernatova, J. Jezek, M. Sery, P. Jakl, et al., "Monitoring

the influence of antibiotic exposure using Raman spectroscopy," in SPIE BiOS,

2014, pp. 89390P-89390P-6.

[66] O. Samek, S. Bernatová, J. Ježek, M. Šiler, M. Šerý, V. Krzyžánek, et al.,

"Identification of individual biofilm-forming bacterial cells using Raman

tweezers," Journal of Biomedical Optics, vol. 20, pp. 051038-051038, 2015.

[67] K. E. Hamden, B. A. Bryan, P. W. Ford, C. Xie, Y.-Q. Li, and S. M. Akula,

"Spectroscopic analysis of Kaposi's sarcoma-associated herpesvirus infected cells

by Raman tweezers," Journal of Virological Methods, vol. 129, pp. 145-151, 2005.

[68] O. F. Dyson, P. W. Ford, D. Chen, Y. Q. Li, and S. M. Akula, "Raman tweezers

provide the fingerprint of cells supporting the late stages of KSHV reactivation,"

Journal of Cellular and Molecular Medicine, vol. 13, pp. 1920-1932, 2009.

[69] K. Chen, Y. Qin, F. Zheng, M. Sun, and D. Shi, "Diagnosis of colorectal cancer

using Raman spectroscopy of laser-trapped single living epithelial cells," Optics

Letters, vol. 31, pp. 2015-2017, 2006.

[70] F. Zheng, Y. Qin, and K. Chen, "Sensitivity map of laser tweezers Raman

spectroscopy for single-cell analysis of colorectal cancer," Journal of Biomedical

Optics, vol. 12, p. 034002, 2007.

[71] J. W. Chan, D. S. Taylor, T. Zwerdling, S. M. Lane, K. Ihara, and T. Huser,

"Micro-Raman spectroscopy detects individual neoplastic and normal

hematopoietic cells," Biophysical Journal, vol. 90, pp. 648-656, 2006.

[72] J. W. Chan, D. S. Taylor, S. M. Lane, T. Zwerdling, J. Tuscano, and T. Huser,

"Nondestructive identification of individual leukemia cells by laser trapping

Raman spectroscopy," Analytical Chemistry, vol. 80, pp. 2180-2187, 2008.


Page 83

[73] J. W. Chan, D. S. Taylor, and D. L. Thompson, "The effect of cell fixation on the

discrimination of normal and leukemia cells with laser tweezers Raman

spectroscopy," Biopolymers, vol. 91, pp. 132-139, 2009.

[74] U. Neugebauer, T. Bocklitz, J. Clement, C. Krafft, and J. Popp, "Towards detection

and identification of circulating tumour cells using Raman spectroscopy," Analyst,

vol. 135, pp. 3178-3182, 2010.

[75] A. Y. Lau, L. P. Lee, and J. W. Chan, "An integrated optofluidic platform for

Raman-activated cell sorting," Lab on a Chip, vol. 8, pp. 1116-1120, 2008.

[76] T. J. Harvey, E. C. Faria, A. Henderson, E. Gazi, A. D. Ward, N. W. Clarke, et al.,

"Spectral discrimination of live prostate and bladder cancer cell lines using Raman

optical tweezers," Journal of Biomedical Optics, vol. 13, p. 064004, 2008.

[77] T. J. Harvey, C. Hughes, A. D. Ward, E. C. Faria, A. Henderson, N. W. Clarke, et

al., "Classification of fixed urological cells using Raman tweezers," Journal of

Biophotonics, vol. 2, pp. 47-69, 2009.

[78] H. nath Banerjee and L. Zhang, "Deciphering the finger prints of brain cancer

astrocytoma in comparison to astrocytes by using near infrared Raman

spectroscopy," Molecular and Cellular Biochemistry, vol. 295, pp. 237-240, 2007.

[79] H. Yao, M. Zhu, G. Wang, L. Peng, B. He, M. He, et al., "Study of Raman spectra

of single carcinoma of nasopharynx cell," Guang Pu Xue Yu Guang Pu Fen Xi,

vol. 27, pp. 1761-1764, 2007.

[80] W.-p. YANG, H.-l. YAO, M. ZHU, G.-w. P. L.-X. WANG, M. HE, and Y.-q. LI,

"Raman Spectrums of Mono-hepatocellular Carcinoma [J]," Laser & Infrared, vol.

9, p. 009, 2007.

[81] H. Yao, Z. Tao, M. Ai, L. Peng, G. Wang, B. He, et al., "Raman spectroscopic

analysis of apoptosis of single human gastric cancer cells," Vibrational

Spectroscopy, vol. 50, pp. 193-197, 2009.

[82] T. J. Moritz, D. S. Taylor, D. M. Krol, J. Fritch, and J. W. Chan, "Detection of

doxorubicin-induced apoptosis of leukemic T-lymphocytes by laser tweezers

Raman spectroscopy," Biomedical Optics Express, vol. 1, pp. 1138-1147, 2010.

[83] R. Liu, Z. Mao, D. L. Matthews, C.-S. Li, J. W. Chan, and N. Satake, "Novel

single-cell functional analysis of red blood cells using laser tweezers Raman

spectroscopy: Application for sickle cell disease," Experimental Hematology, vol.

41, pp. 656-661.e1, 2013.

[84] Z. Tomori, M. Antalik, P. Kesa, J. Kanka, P. Jakl, M. Sery, et al., "Holographic

Raman tweezers controlled by hand gestures and voice commands," Optics and

Photonics Journal, vol. 3, pp. 331-336, 2013.


Page 84

[85] Z. Tomori, J. Kanka, P. Kesa, P. Jakl, M. Sery, S. Bernatova, et al., "Natural user

interface as a supplement of the holographic Raman tweezers," in Proceedings of

SPIE, 2014, pp. 91642P-91642P-8.

[86] M. Boerkamp, T. van Leest, J. Heldens, A. Leinse, M. Hoekman, R. Heideman, et

al., "On-chip optical trapping and Raman spectroscopy using a TripleX dual-

waveguide trap," Optics Express, vol. 22, pp. 30528-30537, 2014.

[87] P. Løvhaugen, B. S. Ahluwalia, T. R. Huser, and O. G. Hellesø, "Serial Raman

spectroscopy of particles trapped on a waveguide," Optics Express, vol. 21, pp.

2964-2970, 2013.

[88] L. Ren, X. Su, Y. Wang, J. Xu, and K. Ning, "QSpec: online control and data

analysis system for single-cell Raman spectroscopy," PeerJ, vol. 2, p. e436, 2014.

[89] S. Wheaton, R. M. Gelfand, and R. Gordon, "Probing the Raman-active acoustic

vibrations of nanoparticles with extraordinary spectral resolution," Nature

Photonics, vol. 9, pp. 68-72, 2014.

[90] R. Gelfand, S. Wheaton, and R. Gordon, "Optical trapping and Raman

spectroscopy of a single MS2 bacteriophage," in Optics in the Life Sciences,

Vancouver, 2015, p. NM4C.5.

[91] f. S. yuan, Y. Huang, J. Zhao, e. shen, Y. Tian, Z. Wu, et al., "Single cell analysis

using laser tweezers Raman spectroscopy, reflectance confocal imaging and

multiphoton fluorescence imaging," in Optics in the Life Sciences, Vancouver,

2015, p. OtT2E.4.

Documents

RAMAN TWEEZERS-An Overview - INFLIBNETshodhganga.inflibnet.ac.in/bitstream/10603/75343/13/13... · 2018-07-08 · RAMAN TWEEZERS-An Overview ... (EVs) of Dictyostelium discoideum,