scholarly journals Label-Free Evaluation of Chromatin Condensation in Human Normal Morphology Sperm Using Raman Spectroscopy

2021 ◽  
Author(s):  
M. Y. Jahmani ◽  
M. E. Hammadeh ◽  
M. A. Al Smadi ◽  
Marko K. Baller
Keyword(s):  
Raman Spectroscopy ◽  
Dna Fragmentation ◽  
Raman Spectra ◽  
Chromatin Condensation ◽  
Function Evaluation ◽  
Label Free ◽  
Chromomycin A3 ◽  
Normal Morphology ◽  
Acridine Orange Staining ◽  
Significant Difference

AbstractChromatin condensation is one of the main factors essential for sperm function. Evaluation of chromatin condensation by current methods render the assessed sperm unsuitable for assisted reproduction. We examined the Raman spectra of normal morphology sperm to determine whether a non-invasive confocal Raman spectroscopy can detect spectral differences between groups having different levels of chromatin condensation. Semen samples from 85 donors who underwent ICSI were obtained. Chromomycin A3, aniline blue and acridine orange staining were performed to evaluate the protamine deficiency, histone retention and DNA fragmentation respectively. Raman spectra were obtained from 50 normal morphology sperm for each donor. Spectral analysis was performed using home written programs in LabVIEW software and samples were grouped based on chromomycin A3 staining. Raman peaks intensities at 670 cm-1, 731 cm-1, 785 cm-1, 858 cm-1, 1062 cm-1, 1098 cm-1, 1185 cm-1, 1372 cm-1, 1424 cm-1, 1450 cm-1, 1532 cm-1, 1618 cm-1 and 1673 cm-1 were significantly correlated with at least one of the sperm staining methods. The median intensity of the Raman peaks at 670 cm-1, 731 cm-1, 785 cm-1, 1062 cm-1, 1098 cm-1, 1185 cm-1, 1372 cm-1, 1424 cm-1, 1450 cm-1, 1532 cm-1, 1618 cm-1 and 1673 cm-1 show a significant difference between the CMA3≤41 and CMA3>41groups. The Raman spectroscopic measurements represent a promising diagnostic tool that has the ability to label-free detect sperm with chromatin abnormalities, such as improper chromatin condensation and DNA fragmentation to a certain degree similar to that of the existing staining techniques at the individual cell level.

Raman spectroscopy-based label-free cell identification using wavelet transform and support vector machine

RSC Advances ◽  
2016 ◽  
Vol 6 (55) ◽  
pp. 50027-50033 ◽  
Author(s):  
S. Bakhtiaridoost ◽  
H. Habibiyan ◽  
S. Muhammadnejad ◽  
M. Haddadi ◽  
H. Ghafoorifard ◽  
...  
Keyword(s):  
Support Vector Machine ◽  
Raman Spectroscopy ◽  
Wavelet Transform ◽  
Raman Spectra ◽  
Free Cell ◽  
Support Vector ◽  
Label Free ◽  
Cell Identification ◽  
Rare Cells

Wavelet transform and SVM applied to Raman spectra makes a powerful and accurate tool for identification of rare cells such as CTCs.

scholarly journals Biochemical Analysis of Leukocytes after In Vitro and In Vivo Activation with Bacterial and Fungal Pathogens Using Raman Spectroscopy

2021 ◽  
Vol 22 (19) ◽  
pp. 10481
Author(s):  
Aikaterini Pistiki ◽  
Anuradha Ramoji ◽  
Oleg Ryabchykov ◽  
Daniel Thomas-Rüddel ◽  
Adrian T. Press ◽  
...  
Keyword(s):  
Raman Spectroscopy ◽  
Single Cell ◽  
Raman Spectra ◽  
Fungal Pathogens ◽  
Infection Model ◽  
Label Free ◽  
Analytical Technique ◽  
Monocytes And Lymphocytes

Biochemical information from activated leukocytes provide valuable diagnostic information. In this study, Raman spectroscopy was applied as a label-free analytical technique to characterize the activation pattern of leukocyte subpopulations in an in vitro infection model. Neutrophils, monocytes, and lymphocytes were isolated from healthy volunteers and stimulated with heat-inactivated clinical isolates of Candida albicans, Staphylococcus aureus, and Klebsiella pneumoniae. Binary classification models could identify the presence of infection for monocytes and lymphocytes, classify the type of infection as bacterial or fungal for neutrophils, monocytes, and lymphocytes and distinguish the cause of infection as Gram-negative or Gram-positive bacteria in the monocyte subpopulation. Changes in single-cell Raman spectra, upon leukocyte stimulation, can be explained with biochemical changes due to the leukocyte’s specific reaction to each type of pathogen. Raman spectra of leukocytes from the in vitro infection model were compared with spectra from leukocytes of patients with infection (DRKS-ID: DRKS00006265) with the same pathogen groups, and a good agreement was revealed. Our study elucidates the potential of Raman spectroscopy-based single-cell analysis for the differentiation of circulating leukocyte subtypes and identification of the infection by probing the molecular phenotype of those cells.

scholarly journals Label-free diagnostics and cancer surgery Raman spectra guidance for the human colon at different excitation wavelengths

RSC Advances ◽  
2019 ◽  
Vol 9 (69) ◽  
pp. 40445-40454 ◽  
Author(s):  
Beata Brozek-Pluska ◽  
Krystian Miazek ◽  
Jacek Musiał ◽  
Radzislaw Kordek
Keyword(s):  
Raman Spectroscopy ◽  
Raman Spectra ◽  
Biological Samples ◽  
Human Colon ◽  
Cancer Surgery ◽  
Label Free ◽  
Minimal Impact ◽  
Structure Sensitive

Raman spectroscopy and imaging are highly structure-sensitive methods that allow the characterization of biological samples with minimal impact.

Label-Free Classification of a Nasopharyngeal Carcinoma Tissue Test at Different Stages Based on Raman Spectroscopy

2017 ◽  
Vol 100 (2) ◽  
pp. 429-433 ◽  
Author(s):  
Mingyu Liu ◽  
Jinyong Lin ◽  
Sufang Qiu ◽  
Weilin Wu ◽  
Gaoqiang Liu ◽  
...  
Keyword(s):  
Raman Spectroscopy ◽  
Nasopharyngeal Carcinoma ◽  
Raman Spectra ◽  
Early Stage ◽  
Support Vector ◽  
Chemical Compositions ◽  
Label Free ◽  
Quick Method ◽  
Clinical Method

Abstract Raman spectroscopy (RS) of nasopharyngeal carcinoma (NPC) tissue provides substantial biomolecular information and various biomedicine features for tissue at different stages of cancer development. This study suggested an automatic and quick method for the classification of Raman spectra at different stages of NPC by multivariate statistical analysis. During RS measurement, Raman spectra were acquired from all NPC tissues in two groups of samples: 30 early-stage NPC patients (stages I and II) and 46 advanced-stage NPC patients (stages III and IV). In addition, a tentative diagnostic algorithm comprising principal components analysis and support vector machine was used to effectively classify multivariate data from the Raman spectra to yield sensitivities (70%; 21 of 30 samples) and specificities (91%; 42 of 46 samples) by the leave-one-out cross-validation method. Meaningful chemical compositions in the classification process were then deduced by analyzing the classified mathematical model. This beneficial work provides a great potential clinical method for the automatic classification of NPC stages and the speculation of the chemical compositions for NPC staging.

scholarly journals Sperm DNA Damage and Protamine Transcripts Content in Iranian Men with Unexplained Infertility

2021 ◽  
Vol In Press (In Press) ◽  
Author(s):  
Mehran Dorostghoal ◽  
Hamid Galehdari ◽  
Masoud Hemadi ◽  
Saba Abdi
Keyword(s):  
Dna Damage ◽  
Dna Fragmentation ◽  
Chromatin Condensation ◽  
Normal Morphology ◽  
Transcript Levels ◽  
Sperm Dna Damage ◽  
Infertile Men ◽  
Sperm Dna ◽  
Polymerase Chain ◽  
Infertile Patients

Background: Incomplete chromatin condensation caused by altered amount of sperm protamines results in DNA fragmentation, which in turn leads to a lack of success in the development of the human embryo. Objectives: This study evaluated the sperm DNA damage and protamine transcripts content in Iranian normozoospermic fertile and infertile men. Methods: DNA damage was analyzed using comet assay. Transcript levels of protamine-1 (PRM1) and protamine-2 (PRM2) in ejaculated spermatozoa were assessed by quantitative real-time polymerase chain reaction (PCR). Results: Significantly higher levels of DNA damage were observed in unexplained infertile men (P = 0.001). DNA fragmentation correlated significantly with sperm total motility (r = -0.413, P = 0.032) and normal morphology (r = -0.424, P = 0.028). PRM1 and PRM2 transcripts contents were significantly lower in normozoospermic infertile men than healthy controls. Sperm PRM1 and PRM2 mRNA ratios were significantly higher (P = 0.035) in unexplained infertile patients than fertile men. Higher DNA damage was found to be significantly associated with reduced transcript levels of PRM1 (r = -0.453, P = 0.018) and PRM2 (r = -0.492, P = 0.009). Protamine transcripts ratios were significantly correlated with sperm normal morphology (r = -0.421, P = 0.029). Conclusions: Our findings showed the prognostic value and clinical utility of the sperm DNA damage and protamine transcripts contents for the discrimination between healthy fertile and unexplained infertile men.

A microfluidic device with integrated impedance detection for λ-DNA

2003 ◽  
Vol 773 ◽  
Author(s):  
Myung-Il Park ◽  
Jonging Hong ◽  
Dae Sung Yoon ◽  
Chong-Ook Park ◽  
Geunbae Im
Keyword(s):  
Microfluidic Device ◽  
Electrochemical Impedance ◽  
Direct Detection ◽  
Full Potential ◽  
Label Free ◽  
Buffer Solution ◽  
Detection Systems ◽  
Significant Difference ◽  
Detection Of Biomolecules ◽  
Impedance Magnitude

AbstractThe large optical detection systems that are typically utilized at present may not be able to reach their full potential as portable analysis tools. Accurate, early, and fast diagnosis for many diseases requires the direct detection of biomolecules such as DNA, proteins, and cells. In this research, a glass microchip with integrated microelectrodes has been fabricated, and the performance of electrochemical impedance detection was investigated for the biomolecules. We have used label-free λ-DNA as a sample biomolecule. By changing the distance between microelectrodes, the significant difference between DW and the TE buffer solution is obtained from the impedance-frequency measurements. In addition, the comparison for the impedance magnitude of DW, the TE buffer, and λ-DNA at the same distance was analyzed.

Submicron Simultaneous IR and Raman Spectroscopy (IR+Raman): Breakthrough Developments in Optical Photothermal IR (O-PTIR) Combined for Enhanced Failure Analysis

2019 ◽  
Author(s):  
Jay Anderson ◽  
Mustafa Kansiz ◽  
Michael Lo ◽  
Curtis Marcott
Keyword(s):  
Raman Spectroscopy ◽  
Failure Analysis ◽  
Data Quality ◽  
Raman Spectra ◽  
Spatial Resolution ◽  
Far Field ◽  
Field Mode ◽  
Microscopic Scale ◽  
Analytical Tools ◽  
Ir And Raman

Abstract Failure analysis of organics at the microscopic scale is an increasingly important requirement, with traditional analytical tools such as FTIR and Raman microscopy, having significant limitations in either spatial resolution or data quality. We introduce here a new method of obtaining Infrared microspectroscopic information, at the submicron level in reflection (far-field) mode, called Optical-Photothermal Infrared (O-PTIR) spectroscopy, that can also generate simultaneous Raman spectra, from the same spot, at the same time and with the same spatial resolution. This novel combination of these two correlative techniques can be considered to be complimentary and confirmatory, in which the IR confirms the Raman result and vice-versa, to yield more accurate and therefore more confident organic unknowns analysis.

scholarly journals Quantitative and Qualitative Analysis of Some Inorganic Compounds by Raman Spectroscopy

1994 ◽  
Vol 48 (7) ◽  
pp. 875-883 ◽  
Author(s):  
Daniel R. Lombardi ◽  
Chao Wang ◽  
Bin Sun ◽  
Augustus W. Fountain ◽  
Thomas J. Vickers ◽  
...  
Keyword(s):  
Aqueous Solution ◽  
Raman Spectroscopy ◽  
Qualitative Analysis ◽  
Raman Spectra ◽  
Solid Phase ◽  
Inorganic Compounds ◽  
Core Material ◽  
Storage Tanks ◽  
Hanford Site ◽  
Sampling Probe

Raman spectra have been measured for a number of nitrates, nitrites, sulfates, ferrocyanides, and ferricyanides, both in the solid phase and in aqueous solution. Accurate locations of peak maxima are given. Limits of detection for some of the compounds are given for solutions and for solid mixtures in NaNO3. Preliminary measurements have been made on core material recovered from the storage tanks on the Hanford site in Richland, Washington. Representative spectra are presented, showing that it is possible to observe responses of individual components from measurements made directly on untreated cores, with the use of a fiberoptic sampling probe.

EXPRESS: Characterization of Drug Resistance in Chronic Myeloid Leukemia Cells Based on Laser Tweezers Raman Spectroscopy

2021 ◽  
pp. 000370282110245
Author(s):  
Qian Zhang ◽  
Minlu Ye ◽  
Lingyan Wang ◽  
Dongmei Jiang ◽  
Shuting Yao ◽  
...  
Keyword(s):  
Raman Spectroscopy ◽  
Chronic Myeloid Leukemia ◽  
Myeloid Leukemia ◽  
K562 Cells ◽  
Laser Tweezers ◽  
Label Free ◽  
Multivariate Statistical ◽  
Analytical Strategy ◽  
Specificity And Sensitivity ◽  
Acid Protein

Multidrug resistance (MDR) is highly associated with poor prognosis of chronic myeloid leukemia (CML). This work aims to explore whether the laser tweezers Raman spectroscopy (LTRS) could be practical in separating adriamycin (ADR) resistance CML cells K562/ADR from its parental cells K562, and to explore the potential mechanisms. Detection of LTRS initially reflected the spectral differences caused by chemoresistance including bands assigned to carbohydrates, amino acid, protein, lipids and nucleic acid. In addition, principal components analysis (PCA) as well as the classification and regression trees (CRT) algorithms showed that the specificity and sensitivity were above 90%. Moreover, the band data-based CRT model and receiver operating characteristic (ROC) curve further determined some important bands and band intensity ratios to be reliable indexes in discriminating K562 chemoresistance status. Finally, we highlighted three metabolism pathways correlated with chemoresistance. This work demonstrates that the label-free LTRS analysis combined with multivariate statistical analyses have great potential to be a novel analytical strategy at the single-cell level for rapid evaluation the chemoresistance status of K562 cells.

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