Confocal Raman microscopy on single living young and old erythrocytes

Kang, Lili; Huang, Yao‐Xiong; Liu, Wenjing; Zheng, Xin-Jing; Wu, Zihao; Luo, Ming

doi:10.1002/bip.21042

Cited by 25 publications

(28 citation statements)

References 39 publications

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“…These peaks correspond to the characteristic bands of deoxyHb[19,26,27], so it suggests that in normal pH condition hemoglobin is mainly deoxygenated. This is consistent with the results of our previous measurement on the Hb in living human erythrocyte[19]. As described previously, the samples were incubated at 4 °C for 72 hours before Raman spectroscopy measurement.…”

Section: Resultsmentioning

confidence: 99%

“…To perform non-disturbance, real-time measurements on the molecular structure and oxygen carrying capacity of hemoglobin in various pH solutions, we also developed a series of novel methods of Raman confocal micro-spectroscopy[19,20]. The Raman spectra measurement was performed using a RAM INV (Horiba JY, France) system with an inverted Olympus optical microscope.…”

Section: Methodsmentioning

confidence: 99%

“…The Raman spectra measurement was performed using a RAM INV (Horiba JY, France) system with an inverted Olympus optical microscope. The 514 nm light from an Ar + iron laser was chosen as the excitation light for it has the advantage in obtaining resonant signal at the characteristic oxidation marker ν4 band (1358cm -1 ) of hemoglobin and providing direct and detailed information about the structure of heme[19–22]. The spectra were recorded between 1800 and 600 cm -1 with a resolution of 1 cm -1 .…”

Section: Methodsmentioning

confidence: 99%

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Pathway and Mechanism of pH Dependent Human Hemoglobin Tetramer-Dimer-Monomer Dissociations

Huang¹,

Wu²,

Huang³

et al. 2013

PLoS ONE

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Hemoglobin dissociation is of great interest in protein process and clinical medicine as well as in artificial blood research. However, the pathway and mechanisms of pH-dependent human Hb dissociation are not clear, whether Hb would really dissociate into monomers is still a question. Therefore, we have conducted a multi-technique investigation on the structure and function of human Hb versus pH. Here we demonstrate that tetramer hemoglobin can easily dissociate into dimer in abnormal pH and the tetramer → dimer dissociation is reversible if pH returns to normal physiological value. When the environmental pH becomes more acidic (<6.5) or alkaline (>8.0), Hb can further dissociate from dimer to monomer. The proportion of monomers increases while the fraction of dimers decreases as pH declines from 6.2 to 5.4. The dimer → monomer dissociation is accompanied with series changes of protein structure thus it is an irreversible process. The structural changes in the dissociated Hbs result in some loss of their functions. Both the Hb dimer and monomer cannot adequately carry and release oxygen to the tissues in circulation. These findings provide a comprehensive understanding on the pH-dependent protein transitions of human Hb, give guideline to explain complex protein processes and the means to control protein dissociation or re-association reaction. They are also of practical value in clinical medicine, blood preservation and blood substitute development.

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Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

See 1 more Smart Citation

Pathway and Mechanism of pH Dependent Human Hemoglobin Tetramer-Dimer-Monomer Dissociations

Huang¹,

Wu²,

Huang³

et al. 2013

PLoS ONE

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“…Raman spectroscopy has been used, for example, to study the mechanisms associated with RBC ageing. 86,87 In these studies, cells were separated by Percoll fractionation (where the lightest 25% were said to be the youngest and the heaviest/densest RBCs were said to be the oldest) and subjected to line mapping analysis (514.2 nm excitation). The intensity of the 1358 cm À1 band (assigned in a previous work to the symmetric half-ring stretch of pyrrole) across the samples suggested that hemoglobin was less uniformly distributed in older RBCs.…”

mentioning

confidence: 99%

Raman Spectroscopy of Blood and Blood Components

et al. 2017

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Blood is a bodily fluid that is vital for a number of life functions in animals. To a first approximation, blood is a mildly alkaline aqueous fluid (plasma) in which a large number of free-floating red cells (erythrocytes), white cells (leucocytes), and platelets are suspended. The primary function of blood is to transport oxygen from the lungs to all the cells of the body and move carbon dioxide in the return direction after it is produced by the cells' metabolism. Blood also carries nutrients to the cells and brings waste products to the liver and kidneys. Measured levels of oxygen, nutrients, waste, and electrolytes in blood are often used for clinical assessment of human health. Raman spectroscopy is a nondestructive analytical technique that uses the inelastic scattering of light to provide information on chemical composition, and hence has a potential role in this clinical assessment process. Raman spectroscopic probing of blood components and of whole blood has been on-going for more than four decades and has proven useful in applications ranging from the understanding of hemoglobin oxygenation, to the discrimination of cancerous cells from healthy lymphocytes, and the forensic investigation of crime scenes. In this paper, we review the literature in the field, collate the published Raman spectroscopy studies of erythrocytes, leucocytes, platelets, plasma, and whole blood, and attempt to draw general conclusions on the state of the field.

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“…To answer these questions, we performed scanning electron microscope (SEM) measurement and confocal Raman micro-spectroscopy on the welded recombinant chromosomes. The reason to use Raman spectroscopy [29] was that the whole chromosome can be studied without physical destruction of the chromatin, thus permitting accurate macromolecular analysis of chromosome structure. Figure 7(A) is the SEM image of the welding joint of the recombinant chromosome.…”

Section: Confocal Raman Micro-spectroscopymentioning

confidence: 99%

Technique of laser chromosome welding for chromosome repair and artificial chromosome creation

Huang¹,

Li²,

Liu³

et al. 2018

Biomed. Opt. Express

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Abstract:Here we report a technique of laser chromosome welding that uses a violet pulse laser micro-beam for welding. The technique can integrate any size of a desired chromosome fragment into recipient chromosomes by combining with other techniques of laser chromosome manipulation such as chromosome cutting, moving, and stretching. We demonstrated that our method could perform chromosomal modifications with high precision, speed and ease of use in the absence of restriction enzymes, DNA ligases and DNA polymerases. Unlike the conventional methods such as de novo artificial chromosome synthesis, our method has no limitation on the size of the inserted chromosome fragment. The inserted DNA size can be precisely defined and the processed chromosome can retain its intrinsic structure and integrity. Therefore, our technique provides a high quality alternative approach to directed genetic recombination, and can be used for chromosomal repair, removal of defects and artificial chromosome creation. The technique may also have applicability on the manipulation and extension of large pieces of synthetic DNA.

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Confocal Raman microscopy on single living young and old erythrocytes

Cited by 25 publications

References 39 publications

Pathway and Mechanism of pH Dependent Human Hemoglobin Tetramer-Dimer-Monomer Dissociations

Pathway and Mechanism of pH Dependent Human Hemoglobin Tetramer-Dimer-Monomer Dissociations

Raman Spectroscopy of Blood and Blood Components

Technique of laser chromosome welding for chromosome repair and artificial chromosome creation

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