Cell Biology JoVE (Open Access) Citable · DOI

Characterization of Sickling During Controlled Automated Deoxygenation with Oxygen Gradient Ektacytometry

Name: Characterization of Sickling During Controlled Automated Deoxygenation with Oxygen Gradient Ektacytometry
Uploaded: 2026-08-06
Duration: 8 min 23 s
Description: In sickle cell disease (SCD), a single point mutation in the gene coding for beta-globin causes the production of abnormal hemoglobin S (HbS). When ...

DOI: 10.3791/60213-v

What you'll learn

✓ Operate oxygen gradient ektacytometer for RBC deformability measurement
✓ Prepare and analyze sickle cell patient blood samples under controlled deoxygenation
✓ Interpret sickling kinetics and treatment response from ektacytometry data
✓ Perform instrument calibration, measurement, and maintenance procedures

Protocol

Biopharma Insights Here, we present oxygen gradient ektacytometry, a rapid and reproducible method to measure red blood cell deformability in samples from patients with sickle cell disease under controlled deoxygenation and reoxygenation. This technique provides a way to study red blood cell sickling and to monitor sickle cell disease treatment efficacy.

Difficulty

advanced

Total time

~45 min per sample (including instrument setup and analysis)

Biosafety

BSL-1

Steps

Start up and calibrate ektacytometer system

Initialize the oxygen gradient ektacytometer, warm up optical and fluidic systems, and perform instrument calibration with reference standards.

▶ 01:12

Collect and prepare blood samples

Obtain blood samples from sickle cell disease patients, perform dilution in isotonic buffer solution, and verify sample readiness for analysis.

▶ 02:13

Execute oxygen gradient ektacytometry measurement

Load prepared sample into ektacytometer chamber and run automated deoxygenation-reoxygenation protocol while recording red blood cell deformability indices across oxygen concentration gradient.

▶ 03:07

Clean and decontaminate ektacytometer

Flush fluidic pathways with deionized water and cleaning solution to remove residual blood cells and maintain optical clarity.

▶ 04:41

Shut down instrument and store safely

Power down ektacytometer systems, secure optical components, and document shutdown completion for next operation.

▶ 05:29

Analyze sickling characterization results

Interpret deformability curves, extract sickling kinetic parameters, and compare treatment efficacy across patient samples using ektacytometry data.

▶ 06:02

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