Characterizations of individual mouse red blood cells parasitized by Babesia microti using 3-D holographic microscopy

Park, Hyun Joo; Hong, Sung-Hee; K, Kim; Cho, Shin‐Hyeong; Lee, Won-Ja; Kim, Young-Chan; Lee, Sang-Eun; Park, YongKeun

doi:10.1038/srep10827

Cited by 87 publications

(64 citation statements)

References 47 publications

(76 reference statements)

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“…Previously, ODT has been widely used to study various biological samples including red blood cells [15][16][17][18][19][20][21][22] however, we were unable to identify lymphocyte cell types due to their indistinguishable cellular morphology and biochemical characteristics.…”

Section: Identification Of Lymphocyte Cell Types Is Crucial For Undermentioning

confidence: 84%

“…Previously, ODT has been widely used to study various biological samples including red blood cells [15][16][17][18][19][20][21][22] , white blood cells (WBC) 23,24 , hepatocytes 25 , cancer cells 16,[26][27][28][29][30][31][32] , neurons 32,33 , bacteria 34,35 , phytoplankton 36 , and hair 37 . In our previous study, we reported that ODT enables the quantitative analysis of WBCs including lymphocytes and macrophages 23 .…”

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confidence: 99%

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Label-free identification of non-activated lymphocytes using three-dimensional refractive index tomography and machine learning

Yoon

Kim

et al. 2017

Preprint

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Identification of lymphocyte cell types is crucial for understanding their pathophysiologic roles in human diseases.Current methods for discriminating lymphocyte cell types primarily relies on labelling techniques with magnetic beads or fluorescence agents, which take time and have costs for sample preparation and may also have a potential risk of altering cellular functions. Here, we present label-free identification of non-activated lymphocyte subtypes using refractive index tomography. From the measurements of three-dimensional refractive index maps of individual lymphocytes, the morphological and biochemical properties of the lymphocytes are quantitatively retrieved. Machine learning methods establish an optimized classification model using the retrieved quantitative characteristics of the lymphocytes to identify lymphocyte subtypes at the individual cell level. We show that our approach enables label-free identification of three lymphocyte cell types (B, CD4+ T, and CD8+ T lymphocytes) with high specificity and sensitivity. The present method will be a versatile tool for investigating the pathophysiological roles of lymphocytes in various diseases including cancers, autoimmune diseases, and virus infections.Lymphocytes consisting of various cell types including B, helper (CD4+) T, cytotoxic (CD8+) T, and regulatory T lymphocytes, and play crucial roles in the adaptive immune system 1 . Each lymphocyte cell type has different functions: B lymphocytes produce antibodies, and T lymphocytes recognize a specific antigen and execute effector functions. The lymphocyte population and function are tightly regulated to defend the host against harmful invaders or abnormal conditions 1,2 . Disturbances in lymphocyte function and regulation are related to various diseases including cancers 3-5 , autoimmune diseases 6,7 , and virus infections 8,9 .. CC-BY 4.0 International license not peer-reviewed) is the author/funder. It is made available under a The copyright holder for this preprint (which was . http://dx.doi.org/10.1101/107805 doi: bioRxiv preprint first posted online Feb. 11, 2017; To understand the roles of different types of lymphocytes, several methods based on labelling techniques have been developed to identify and discriminate lymphocyte cell types. Because different kinds of lymphocytes have very similar cellular morphology such as a large nucleus with small cytosolic regions and round shapes, conventional optical methods such as bright-field microscopy or phase contrast microscopy have limited ability in classifying lymphocyte cell types 10 .To overcome this, a specific surface membrane proteins, known as surface markers, are recognized and tagged with magnetic beads or fluorescence molecules via antigen-antibody binding. And then, specific types of lymphocytes are identified and separated by magnetic forces or fluorescence signals 11 . Targeting surface markers is a precise and efficient manner to determine the cell types; however, labelling methods have potential risks of altering cellular functions b...

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Section: Identification Of Lymphocyte Cell Types Is Crucial For Undermentioning

confidence: 84%

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confidence: 99%

Label-free identification of non-activated lymphocytes using three-dimensional refractive index tomography and machine learning

Yoon

Kim

et al. 2017

Preprint

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“…According to the literature Babesia microti penetrates directly into erythrocytes without the stages of initial proliferation at other organs [5,21].This feature is related to the specifi city of the impact of the membrane of red blood cells with a particular species of piroplasm [22,23].…”

Section: Discussionmentioning

confidence: 99%

Babesia Microti – Known and Unknown Protists

Albertyńska¹,

Rupik²,

Hermyt³

et al. 2017

Glob J Zool

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B. microti is known as the main etiological agent of human babesiosis and there are some case studies for that disease, highlighting the fact that this is an important "emerging tick-borne disease". However a lot of information about this protist is unclear.The reactions in the liver are noticeable already after the fi rst three weeks of infection. This therefore provides the basis for discussion of the effect of B. microti contact with hepatocytes in vitro and in vivo. This issue is an essential objective of our study.The effect of B. microti on the liver as an organ and the individual hepatocytes is evident. This may be an indirect impact, through the various factors of metabolic pathways. Ultrastructural studies of breeding co-culture of hepatocytes and B. microti has shown, however, that piroplasms B. microti are adhering to the outer surface of the hepatocytes. This contact is associated with destructive changes in hepatocytes both necrotic and apoptotic.

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“…Similarly, other RBC infecting microbes can be investigated, such as Babesia microti, which is an obligate parasite in humans, hamster, and mouse. To find alternatives to methods requiring experience of professional technicians, usually based on optical microscope with Giemsa-stained blood smears, DH methodology was exploited [50]. The authors found the technique to be useful for determining morphological modifications in host RBCs, quantifying contents, and concentration of the cellular dry mass, as well as dynamic membrane fluctuations measured at the individual cell level, which are in turn strongly correlated with the mechanical deformability of cell membrane.…”

Section: Blood Cell Screeningmentioning

confidence: 99%

Holography: The Usefulness of Digital Holographic Microscopy for Clinical Diagnostics

El-Schich¹,

Kamlund²,

Janicke³

et al. 2017

Holographic Materials and Optical Systems

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Digital holographic (DH) microscopy is a digital high-resolution holographic imaging technique with the capacity of quantification of cellular conditions without any staining or labeling of cells. The unique measurable parameters are the cell number, cell area, thickness, and volume, which can be coupled to proliferation, migration, cell cycle analysis, viability, and cell death. The technique is cell friendly, fast and simple to use and has unique imaging capabilities for time-lapse investigations on both the single cell and the cell-population levels. The interest for analyzing specifically cell volume changes with DH microscopy, resulting from cytotoxic treatments, drug response, or apoptosis events has recently increased in popularity. We and others have used DH microscopy showing that the technique has the sensitivity to distinguish between different cells and treatments. Recently, DH microscopy has been used for cellular diagnosis in the clinic, providing support for using the concept of DH, e.g., screening of malaria infection of red blood cells (RBC), cervix cancer screening, and sperm quality. Because of its quick and label-free sample handling, DH microscopy will be an important tool in the future for personalized medicine investigations, determining the optimal therapeutic concentration for both different cancer types and individual treatments.

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Characterizations of individual mouse red blood cells parasitized by Babesia microti using 3-D holographic microscopy

Cited by 87 publications

References 47 publications

Label-free identification of non-activated lymphocytes using three-dimensional refractive index tomography and machine learning

Label-free identification of non-activated lymphocytes using three-dimensional refractive index tomography and machine learning

Babesia Microti – Known and Unknown Protists

Holography: The Usefulness of Digital Holographic Microscopy for Clinical Diagnostics

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