Dataset Information

Optical Investigation of Individual Red Blood Cells for Determining Cell Count and Cellular Hemoglobin Concentration in a Microfluidic Channel.

ABSTRACT: We demonstrate a blood analysis routine by observing red blood cells through light and digital holographic microscopy in a microfluidic channel. With this setup a determination of red blood cell (RBC) concentration, the mean corpuscular volume (MCV), and corpuscular hemoglobin concentration mean (CHCM) is feasible. Cell count variations in between measurements differed by 2.47% with a deviation of -0.26×106 μL to the reference value obtained from the Siemens ADVIA 2120i. Measured MCV values varied by 2.25% and CHCM values by 3.78% compared to the reference ADVIA measurement. Our results suggest that the combination of optical analysis with microfluidics handling provides a promising new approach to red blood cell counts.

SUBMITTER: Reichenwallner AK

PROVIDER: S-EPMC8066749 | biostudies-literature | 2021 Mar

REPOSITORIES: biostudies-literature

ACCESS DATA

Publications

Optical Investigation of Individual Red Blood Cells for Determining Cell Count and Cellular Hemoglobin Concentration in a Microfluidic Channel.

Reichenwallner Ann-Kathrin AK Vurmaz Esma E Battis Kristina K Handl Laura L Üstün Helin H Mach Tivadar T Hörnig Gabriele G Lipfert Jan J Richter Lukas L

Micromachines 20210326 4

We demonstrate a blood analysis routine by observing red blood cells through light and digital holographic microscopy in a microfluidic channel. With this setup a determination of red blood cell (RBC) concentration, the mean corpuscular volume (MCV), and corpuscular hemoglobin concentration mean (CHCM) is feasible. Cell count variations in between measurements differed by 2.47% with a deviation of -0.26×106 μL to the reference value obtained from the Siemens ADVIA 2120i. Measured MCV values vari ...[more]

PMID: 33810262

Similar Datasets

Project description:During its intraerythrocytic asexual reproduction cycle Plasmodium falciparum consumes up to 80% of the host cell hemoglobin, in large excess over its metabolic needs. A model of the homeostasis of falciparum-infected red blood cells suggested an explanation based on the need to reduce the colloid-osmotic pressure within the host cell to prevent its premature lysis. Critical for this hypothesis was that the hemoglobin concentration within the host cell be progressively reduced from the trophozoite stage onwards.The experiments reported here were designed to test this hypothesis by direct measurements of the hemoglobin concentration in live, infected red cells. We developed a novel, non-invasive method to quantify the hemoglobin concentration in single cells, based on Förster resonance energy transfer between hemoglobin molecules and the fluorophore calcein. Fluorescence lifetime imaging allowed the quantitative mapping of the hemoglobin concentration within the cells. The average fluorescence lifetimes of uninfected cohorts was 270+/-30 ps (mean+/-SD; N = 45). In the cytoplasm of infected cells the fluorescence lifetime of calcein ranged from 290+/-20 ps for cells with ring stage parasites to 590+/-13 ps and 1050+/-60 ps for cells with young trophozoites and late stage trophozoite/early schizonts, respectively. This was equivalent to reductions in hemoglobin concentration spanning the range from 7.3 to 2.3 mM, in line with the model predictions. An unexpected ancillary finding was the existence of a microdomain under the host cell membrane with reduced calcein quenching by hemoglobin in cells with mature trophozoite stage parasites.The results support the predictions of the colloid-osmotic hypothesis and provide a better understanding of the homeostasis of malaria-infected red cells. In addition, they revealed the existence of a distinct peripheral microdomain in the host cell with limited access to hemoglobin molecules indicating the concentration of substantial amounts of parasite-exported material.

Project description:BackgroundHematocrit measurement has been an indispensable tool for monitoring plasma leakage and bleeding in dengue patients. However, hematocrit measurement by automated methods is hampered by frequent venipunctures. Utility of point-of-care hemoglobin (POC-Hb) test for monitoring dengue patients has not been established. We evaluated the relationship between hemoglobin measured by POC-Hb testing and hematocrit measured by the automated method in adult dengue patients.Methodology and principal findingsAdult dengue patients were recruited at two university hospitals in Thailand from October 2019 to December 2020. POC-Hb test was performed using capillary blood simultaneously with venipuncture to obtain whole blood for an automated complete blood count (CBC) analysis. The correlation of hemoglobin and hematocrit measurement was evaluated. A total of 44 dengue patients were enrolled. Twenty-nine patients (65.9%) were female, with a median age of 31 years (interquartile range 22-41). Of the enrolled patients, 30 (68.2%), 11 (25.0%), and 3 (6.8%) were classified as dengue without warning signs, with warning signs, and severe dengue, respectively. Seven patients (15.9%) had hemoconcentration, and five patients (11.3%) had bleeding. A total of 216 pairs of POC-Hb and CBC were evaluated. A significant positive correlation was observed between hemoglobin measured by POC-Hb testing and hematocrit measured by an automated CBC (r = 0.869, p <0.001). Bland-Altman analysis between hemoglobin measured by POC-Hb testing and an automated CBC showed a bias of -0.43 (95% limit of agreement of -1.81 and 0.95). Using the cutoff of POC-Hb ≥20% as a criteria for hemoconcentration, the sensitivity and specificity of hemoconcentration detected by POC-Hb device were 71.4% and 100.0%, respectively.ConclusionsHemoglobin measurement by POC-Hb testing has a strong correlation with hematocrit in adult patients with dengue fever. However, the sensitivity in detecting hemoconcentration is fair. The adjunct use of capillary POC-Hb testing can decrease the frequency of venipuncture. Further study in children is encouraged.

Dataset Information

Optical Investigation of Individual Red Blood Cells for Determining Cell Count and Cellular Hemoglobin Concentration in a Microfluidic Channel.

Publications

Optical Investigation of Individual Red Blood Cells for Determining Cell Count and Cellular Hemoglobin Concentration in a Microfluidic Channel.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets