Characterizing stored red blood cells using ultra-high throughput holographic cytometry

AbstractHolographic cytometry is introduced as an ultra-high throughput implementation of quantitative phase image based on off-axis interferometry of cells flowing through parallel microfluidic channels. Here, it is applied for characterizing morphological changes of red blood cells during storage under regular blood bank condition. The approach allows high quality phase imaging of a large number of cells greatly extending our ability to study cellular phenotypes using individual cell images. Holographic cytology measurements show multiple physical traits of the cells, including optical volume and area, which are observed to consistently change over the storage time. In addition, the large volume of cell imaging data can serve as training data for machine learning algorithms. For the study here, logistic regression is used to classify the cells according to the storage time points. The results of the classifiers demonstrate the potential of holographic cytometry as a diagnostic tool.

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Single Cell Analysis of Stored Red Blood Cells Using Ultra-High Throughput Holographic Cytometry

Cells ◽

10.3390/cells10092455 ◽

2021 ◽

Vol 10 (9) ◽

pp. 2455

Author(s):

Han-Sang Park ◽

Hillel Price ◽

Silvia Ceballos ◽

Jen-Tsan Chi ◽

Adam Wax

Keyword(s):

Red Blood Cells ◽

High Throughput ◽

Blood Cells ◽

Storage Time ◽

Single Cells ◽

Machine Learning Algorithms ◽

Training Data ◽

Microfluidic Channels ◽

Phase Imaging ◽

Time Points

Holographic cytometry is introduced as an ultra-high throughput implementation of quantitative phase imaging of single cells flowing through parallel microfluidic channels. Here, the approach was applied for characterizing the morphology of individual red blood cells during storage under regular blood bank conditions. Samples from five blood donors were examined, over 100,000 cells examined for each, at three time points. The approach allows high-throughput phase imaging of a large number of cells, greatly extending our ability to study cellular phenotypes using individual cell images. Holographic cytology images can provide measurements of multiple physical traits of the cells, including optical volume and area, which are observed to consistently change over the storage time. In addition, the large volume of cell imaging data can serve as training data for machine-learning algorithms. For the study here, logistic regression was used to classify the cells according to the storage time points. The analysis showed that at least 5000 cells are needed to ensure accuracy of the classifiers. Overall, results showed the potential of holographic cytometry as a diagnostic tool.

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Time Dependent Assessment of Morphological Changes: Leukodepleted Packed Red Blood Cells Stored in SAGM

BioMed Research International ◽

10.1155/2016/4529434 ◽

2016 ◽

Vol 2016 ◽

pp. 1-7 ◽

Cited By ~ 12

Author(s):

Ibrahim Mustafa ◽

Asma Al Marwani ◽

Khuloud Mamdouh Nasr ◽

Noora Abdulla Kano ◽

Tameem Hadwan

Keyword(s):

Red Blood Cells ◽

Blood Cells ◽

Storage Time ◽

Morphological Changes ◽

Storage Period ◽

Osmotic Fragility ◽

Prolonged Storage ◽

Morphological Alterations ◽

Packed Red Blood Cells ◽

Stored Blood

Usually packed red blood cells (pRBCs) require specific conditions in storage procedures to ensure the maximum shelf life of up to 42 days in 2–6°C. However, molecular and biochemical consequences can affect the stored blood cells; these changes are collectively labeled as storage lesions. In this study, the effect of prolonged storage was assessed through investigating morphological changes and evaluating oxidative stress. Samples from leukodepleted pRBC in SAGM stored at 4°C for 42 days were withdrawn aseptically on day 0, day 14, day 28, and day 42. Morphological changes were observed using scanning electron microscopy and correlated with osmotic fragility and hematocrit. Oxidative injury was studied through assessing MDA level as a marker for lipid peroxidation. Osmotic fragility test showed that extended storage time caused increase in the osmotic fragility. The hematocrit increased by 6.6% from day 0 to day 42. The last 2 weeks show alteration in the morphology with the appearance of echinocytes and spherocytes. Storage lesions and morphological alterations appeared to affect RBCs during the storage period. Further studies should be performed to develop strategies that will aid in the improvement of stored pRBC quality and efficacy.

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An Evaluation of Morphological Changes and Deformability of Suspended Red Blood Cells Prepared Using Whole Blood with Different Hemoglobin Levels of Tibetans

Transfusion Medicine and Hemotherapy ◽

10.1159/000513319 ◽

2021 ◽

pp. 1-10

Author(s):

Rui Zhong ◽

Dingding Han ◽

Xiaodong Wu ◽

Hong Wang ◽

Wanjing Li ◽

...

Keyword(s):

Red Blood Cells ◽

Whole Blood ◽

Blood Cells ◽

Morphological Changes ◽

Osmotic Fragility ◽

Spherical Shape ◽

Hypoxic Environment ◽

Wide Range ◽

Group A ◽

Cell Membrane Protein

Background: The hypoxic environment stimulates the human body to increase the levels of hemoglobin (HGB) and hematocrit and the number of red blood cells. Such enhancements have individual differences, leading to a wide range of HGB in Tibetans’ whole blood (WB). Study Design: WB of male Tibetans was divided into 3 groups according to different HGB (i.e., A: >120 but ≤185 g/L, B: >185 but ≤210 g/L, and C: >210 g/L). Suspended red blood cells (SRBC) processed by collected WB and stored in standard conditions were examined aseptically on days 1, 14, 21, and 35 after storage. The routine biochemical indexes, deformability, cell morphology, and membrane proteins were tested. Results: Mean corpuscular volume, adenosine triphosphate, pH, and deformability were not different in group A vs. those in storage (p > 0.05). The increased rate of irreversible morphology of red blood cells was different among the 3 groups, but there was no difference in the percentage of red blood cells with an irreversible morphology after 35 days of storage. Group C performed better in terms of osmotic fragility and showed a lower rigid index than group A. Furthermore, SDS-PAGE revealed similar cross-linking degrees of cell membrane protein but the band 3 protein of group C seemed to experience weaker clustering than that of group A as detected by Western Blot analysis after 35 days of storage. Conclusions: There was no difference in deformability or morphological changes in the 3 groups over the 35 days of storage. High HGB levels of plateau SRBC did not accelerate the RBC change from a biconcave disc into a spherical shape and it did not cause a reduction in deformability during 35 days of preservation in bank conditions.

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Sonoporation enables high-throughput loading of trehalose into red blood cells

Cryobiology ◽

10.1016/j.cryobiol.2020.12.005 ◽

2020 ◽

Author(s):

Brett R. Janis ◽

Mariah C. Priddy ◽

Meghan R. Otto ◽

Jonathan A. Kopechek ◽

Michael A. Menze

Keyword(s):

Red Blood Cells ◽

High Throughput ◽

Blood Cells

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Hemoglobin Oxidation in Stored Blood Accelerates Hemolysis and Oxidative Injury to Red Blood Cells

Journal of Laboratory Physicians ◽

10.1055/s-0040-1721156 ◽

2020 ◽

Vol 12 (04) ◽

pp. 244-249

Author(s):

Ibrahim Mustafa ◽

Tameem Ali Qaid Hadwan

Keyword(s):

Red Blood Cells ◽

Blood Cells ◽

Morphological Changes ◽

Hemoglobin Concentration ◽

Red Cell Membrane ◽

Rbc Membrane ◽

Hemoglobin Oxidation ◽

Rbc Indices ◽

Methemoglobin Formation ◽

Stored Blood

Abstract Introduction Maintaining blood supply is a challenge in blood banks. Red blood cells (RBCs) stored at 4°C experience issues of biochemical changes due to metabolism of cells, leading to changes collectively referred to as “storage lesions.” Oxidation of the red cell membrane, leading to lysis, contributes to these storage lesions. Methods Blood bags with CPD-SAGM stored at 4°C for 28 days were withdrawn aseptically on days 1, 14, and 28. Hematology analyzer was used to investigate RBC indices. Hemoglobin oxidation was studied through spectrophotometric scan of spectral change. RBC lysis was studied with the help of Drabkin's assay, and morphological changes were observed by light and scan electron microscopy. Results RBCs show progressive changes in morphology echinocytes and spherocytes on day 28. There was 0.85% RBC lysis, an approximately 20% decrease in percentage oxyhemoglobin, and a 14% increase in methemoglobin formation, which shows hemoglobin oxidation on day 28. Conclusions Oxidative damage to RBC, with an increase in storage time was observed in the present study. The observed morphological changes to RBC during the course of increased time shows that there is progressive damage to RBC membrane and a decrease in hemoglobin concentration; percentage RBC lysis is probably due to free hemoglobin and iron.

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High-throughput assessment of mechanical properties of stem cell derived red blood cells, toward cellular downstream processing

Scientific Reports ◽

10.1038/s41598-017-14958-w ◽

2017 ◽

Vol 7 (1) ◽

Cited By ~ 13

Author(s):

Ewa Guzniczak ◽

Maryam Mohammad Zadeh ◽

Fiona Dempsey ◽

Melanie Jimenez ◽

Henry Bock ◽

...

Keyword(s):

Mechanical Properties ◽

Stem Cell ◽

Red Blood Cells ◽

High Throughput ◽

Blood Cells ◽

Downstream Processing

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Using Redox Potential as a Feasible Marker for Banked Blood Quality and the State of Oxidative Stress in Stored Red Blood Cells

10.21203/rs.3.rs-104378/v1 ◽

2020 ◽

Author(s):

Rodney C Daniels ◽

Hyesun Jun ◽

Robertson D Davenport ◽

Maryanne M Collinson ◽

Kevin R Ward

Keyword(s):

Oxidative Stress ◽

Redox Potential ◽

Red Blood Cells ◽

Healthy Volunteers ◽

Blood Cells ◽

Storage Time ◽

Cell Function ◽

Venous Blood ◽

Functional Changes ◽

Over Time

Abstract Background Stored Red Blood Cells (RBCs) may undergo oxidative stress over time, with functional changes affecting critical tasks such as oxygen delivery. Central to these changes are oxidation-reduction (redox) reactions and the redox potential (RP) that must be maintained for proper cell function. RP imbalance can lead to oxidative stress that may contribute to storage lesions and transfusion-related morbidities. Direct measures of RP may allow for evaluation of erythrocyte quality and enable corrections of RP prior to transfusion. Methods Multiple random RBC segments were tested, ranging in age from 5 to 40 days at 5 day intervals. RP was recorded by measuring open circuit potential of RBCs using novel nanoporous gold electrodes with Ag/AgCl reference. RP measures were also performed on peripheral venous blood samples from 10 healthy volunteers. RP measures were compared between groups of aged RBCs, and with volunteer blood. Results Stored RBCs show time-dependent increases in RP. There were significant differences in Day 5 RP compared to all other groups (p≤0.005), Day 10-15 vs ages ≥ Day 20 (p≤0.025), Day 20-25 vs Day 40 (p=0.039), and all groups compared to healthy volunteers. RP became more positive over time suggesting ongoing oxidation as RBCs age. However, storage time alone does not predict the ultimate RP value measured from a given unit.Conclusions There are significant differences in RP between freshly stored RBCs and all others, with RP becoming more positive over time. However, storage time alone does not predict RP, indicating RP screening may be important independent of storage time and may serve as a marker of RBC quality and state of oxidative stress. RP measurements may also provide a target by which to restore RP balance in aged pRBCs, improving their clinical effectiveness while reducing associated morbidities.

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