Application of image flow cytometry and photoacoustics for the characterization of red blood cell storage lesions

Significant functional/structural changes of red blood cells (RBCs) have been documented during its in vitro storage. Collectively termed as RBC storage lesions, changes include an increase in RBC oxygen saturation (SO2) and an increase in irreversibly damaged RBCs (spheroechinocytes). In this work, novel optical techniques are presented for determining the spheroechinocyte population as a function of storage time via automated image flow cytometry (IFC) morphology characterization, and the acquisition of RBC SO2 via an in situ photoacoustic (PA) method. Blood gas analysis (BGA) was used as the gold standard SO2 measure. Over the lifespan of seven blood bags, the IFC spheroechinocyte population – PA SO2 correlation was found to be strong (0.600.95) shows high potential for in situ monitoring of RBC storage lesions.

Application of image flow cytometry and photoacoustics for the characterization of red blood cell storage lesions

Significant functional/structural changes of red blood cells (RBCs) have been documented during its in vitro storage. Collectively termed as RBC storage lesions, changes include an increase in RBC oxygen saturation (SO2) and an increase in irreversibly damaged RBCs (spheroechinocytes). In this work, novel optical techniques are presented for determining the spheroechinocyte population as a function of storage time via automated image flow cytometry (IFC) morphology characterization, and the acquisition of RBC SO2 via an in situ photoacoustic (PA) method. Blood gas analysis (BGA) was used as the gold standard SO2 measure. Over the lifespan of seven blood bags, the IFC spheroechinocyte population – PA SO2 correlation was found to be strong (0.600.95) shows high potential for in situ monitoring of RBC storage lesions.

High-throughput enrichment and isolation of megakaryocyte progenitor cells from the mouse bone marrow

Megakaryocytes are a rare population of cells that develop in the bone marrow and function to produce platelets that circulate throughout the body and form clots to stop or prevent bleeding. A major challenge in studying megakaryocyte development, and the diseases that arise from their dysfunction, is the identification, classification, and enrichment of megakaryocyte progenitor cells that are produced during hematopoiesis. Here, we present a high throughput strategy for identifying and isolating megakaryocytes and their progenitor cells from a heterogeneous population of bone marrow samples. Specifically, we couple thrombopoietin (TPO) induction, image flow cytometry, and principal component analysis (PCA) to identify and enrich for megakaryocyte progenitor cells that are capable of self-renewal and directly differentiating into mature megakaryocytes. This enrichment strategy distinguishes megakaryocyte progenitors from other lineage-committed cells in a high throughput manner. Furthermore, by using image flow cytometry with PCA, we have identified a combination of markers and characteristics that can be used to isolate megakaryocyte progenitor cells using standard flow cytometry methods. Altogether, these techniques enable the high throughput enrichment and isolation of cells in the megakaryocyte lineage and have the potential to enable rapid disease identification and diagnoses ahead of severe disease progression.