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Label-free blood typing by Raman spectroscopy (RS) is demonstrated by training an artificial intelligence (AI) model on 271 blood typed donor whole blood samples. A fused silica micro-capillary flow cell enables fast generation of a large dataset of Raman spectra of individual donors. A combination of resampling methods, machine learning and deep learning is used to classify the ABO blood group, 27 erythrocyte antigens, 4 platelet antigens, regular anti-B titers of blood group A donors, regular anti-A,-B titers of blood group O donors, and ABH-secretor status, from a single Raman spectrum. The average area under the curve value of the ABO classification is 0.91 ± 0.03 and 0.72 ± 0.09, respectively, for the remaining traits. The classification performance of all parameters is discussed in the context of dataset balance and antigen concentration. Post-hoc scalability analysis of the models shows the potential of RS and AI for future applications in transfusion medicine and blood banking., Label-free blood typing by Raman spectroscopy (RS) is demonstrated by training an artificial intelligence (AI) model on 271 blood typed donor whole blood samples. A fused silica micro-capillary flow cell enables fast generation of a large dataset of Raman spectra of individual donors. A combination of resampling methods, machine learning and deep learning is used to classify the ABO blood group, 27 erythrocyte antigens, 4 platelet antigens, regular anti-B titers of blood group A donors, regular anti-A,-B titers of blood group O donors, and ABH-secretor status, from a single Raman spectrum. The average area under the curve value of the ABO classification is 0.91 ± 0.03 and 0.72 ± 0.09, respectively, for the remaining traits. The classification performance of all parameters is discussed in the context of dataset balance and antigen concentration. Post-hoc scalability analysis of the models shows the potential of RS and AI for future applications in transfusion medicine and blood banking.

A compact microfluidic flow cytometer is demonstrated, comprising viscoelastic flow focusing in fused silica capillaries and a fiber optical interface. Viscoelastic flow focusing enables simple device design and operation with a single-inlet/outlet fluidic configuration. Fused silica capillaries with different inner diameters are effortlessly interchanged to eliminate blockage ratio limitations and enable single-train particle focusing for a wide range of particle sizes and geometries. The compact system is mounted on an inverted microscope for easy integration with optical imaging and other optofluidic modalities, such as optical trapping and particle sorting. A real-time cytometric analysis of three channels, forward scattering, side scattering, and fluorescence detection, is performed on LABVIEW. A throughput of 3500 events s(-1) is performed on particles of sizes ranging from 2 to 20 mu m, using capillaries of different inner diameters ranging from 30 to 75 mu m. The outer diameter of all capillaries is identical to the cladding diameter of the applied optical fibers. This enables easy exchange and precise optical alignment of fibers and capillaries on a microfabricated jig. The performance of the microfluidic flow cytometer is benchmarked using polystyrene calibration beads, poly(lactic-co-glycolic acid) particles, erythrocytes, THP-1 leukemic monocytes, and human metaphase chromosomes.

ABO blood typing is the determination of four different blood groups: type A, B, AB, or O. Clinically approved ABO blood typing methods are suffering from expensive reagents and multiple time-consuming cross-referencing steps, creating the need for fast, sustainable, sensitive, and label-free technologies. Raman spectroscopy techniques have shown potential to distinguish biomolecules and blood components such as purified serum proteins, albumin, and globulin. In combination with machine learning tools, the accuracy and specificity of Raman spectroscopic measurements can be improved and adapted to clinical applications. This study presents a multivariate analysis of human-blood samples for ABO blood typing using Raman spectroscopy and support vector machine (SVM) classification. A custom-built NIR Raman spectroscopy setup with a 785 nm wavelength laser is coupled into an inverted microscope to collect Raman spectra from each blood sample. Donor samples are drawn from EDTA tubes into a fused silica microcapillary without dilution and sample preparation steps. Raman measurements from more than 270 donor samples are analyzed to get accurate blood typing predictions. The blood types are distinguished pairwise by an average AUC score of 0.94, showing great potential of the developed system for future blood typing applications.

Inborn hemolytic anemia requiring frequent blood transfusions can be a life-threatening disease. Treatment, besides blood transfusion, includes iron chelation for prevention of iron accumulation due to frequent blood transfusions. We present the results of a clinical investigation where the proband was diagnosed with severe hemolytic anemia of unknown origin soon after birth. Transfusion was required every 4-6 weeks. After whole exome sequencing of the proband and his parents as well as a healthy sibling, we established that the proband had a compound heterozygous state carrying two rare variants in the erythrocytic spectrin gene, SPTA1. The maternal allele was a stop mutation (rs755630903) and the paternal allele was a missense mutation (rs375506528). The healthy sibling had the paternal variant but not the maternal variant. These rare variants of SPTA1 most likely account for the hemolytic anemia. A severely reduced osmotic resistance in the erythrocytes from the proband was demonstrated. Splenectomy considerably improved the hemolytic anemia and obviated the need for blood transfusion despite the severe clinical presentation.

Background: Laboratory monitoring of mother, fetus, and newborn in hemolytic disease of fetus and newborn (HDFN) aims to guide clinicians and the immunized women to focus on the most serious problems of alloimmunization and thus minimize the consequences of HDFN in general and of anti-D in particular. Here, we present the current approach of laboratory screening and testing for prevention and monitoring of HDFN at the Copenhagen University Hospital in Denmark. Summary: All pregnant women are typed and screened in the 1st trimester. This serves to identify the RhD-negative pregnant women who at gestational age (GA) of 25 weeks are offered a second screen test and a non-invasive fetal RhD prediction. At GA 29 weeks, and again after delivery, non-immunized RhD-negative women carrying an RhD-positive fetus are offered Rh immunoglobulin. If the 1st trimester screen reveals an alloantibody, antenatal investigation is initiated. This also includes RhD-positive women with alloantibodies. Specificity and titer are determined, the fetal phenotype is predicted by non-invasive genotyping based on cell-free DNA (RhD, K, Rhc, RhC, RhE, ABO), and serial monitoring of titer commences. Based on titers and specificity, monitoring with serial peak systolic velocity measurements in the fetal middle cerebral artery to detect anemia will take place. Intrauterine transfusion is given when fetal anemia is suspected. Monitoring of the newborn by titer and survival of fetal red blood cells by flow cytometry will help predict the length of the recovery of the newborn.