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Characterization of red blood cells with multiwavelength transmission spectroscopy.

Serebrennikova YM, Huffman DE, Garcia-Rubio LH - Biomed Res Int (2015)

Bottom Line: The MWT spectra were quantitatively analyzed with a Mie theory based interpretation model modified to incorporate the effects of the nonsphericity and orientation of RBCs.The changes in the RBC volume, surface area, aspect ratio, and hemoglobin composition were used to trace the morphological and compositional alterations in the infected RBCs occurring with parasites' development and to provide insights into parasite-host interactions.The MWT method was shown to be reliable for determination of the RBC morphological parameters and to be valuable for identification of the RBC pathologic changes and disease states.

View Article: PubMed Central - PubMed

Affiliation: Claro Scientific, LLC., 10100 Dr. Martin Luther King Jr. Street N., St. Petersburg, FL 33716, USA.

ABSTRACT
Multiwavelength transmission (MWT) spectroscopy was applied to the investigation of the morphological parameters and composition of red blood cells (RBCs). The MWT spectra were quantitatively analyzed with a Mie theory based interpretation model modified to incorporate the effects of the nonsphericity and orientation of RBCs. The MWT spectra of the healthy and anemic samples were investigated for the RBC indices in open and blinded studies. When MWT performance was evaluated against a standard reference system, very good agreement between two methods, with R (2) > 0.85 for all indices studied, was demonstrated. The RBC morphological parameters were used to characterize three types of anemia and to draw an association between RBC morphology and anemia severity. The MWT spectra of RBCs infected with malaria parasite Plasmodium falciparum at different life cycle stages were analyzed for RBC morphological parameters. The changes in the RBC volume, surface area, aspect ratio, and hemoglobin composition were used to trace the morphological and compositional alterations in the infected RBCs occurring with parasites' development and to provide insights into parasite-host interactions. The MWT method was shown to be reliable for determination of the RBC morphological parameters and to be valuable for identification of the RBC pathologic changes and disease states.

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Related in: MedlinePlus

Histogram distribution plots of MCV (fL) index for samples from healthy donors (a) and anemic patients (b).
© Copyright Policy - open-access
Related In: Results  -  Collection


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fig4: Histogram distribution plots of MCV (fL) index for samples from healthy donors (a) and anemic patients (b).

Mentions: The MCV distribution for samples from healthy donors approached normal as can be seen in Figure 4(a). On the other hand, the distribution of the MCV values for the samples from anemic patients was more complex with side peaks at low and high MCV ranges (Figure 4(b)). Normocytic anemia, that is, with MCV values within 80–100 fL range, in the case of cancer patients is likely to be indicative of the bone marrow failure [3] and, depending on a variety of factors, including the type and intensity of chemotherapy, occurs in over 60% of cancer patients [18–20]. The bone marrow failure resulted in decreased production of functional RBCs and coupled with cancer related inflammatory response [18] and/or chemotherapy induced [19, 20] increase in destruction of RBCs; blood loss leads to decrease in HGB, RBC count, and HCT. Although hemolysis can also be associated with normocytic anemia, it was not detected in any sample of this study. The peak at smaller MCV values in Figure 4(b) was indicative of microcytic anemia, that is, with MCV values below 80 fL, which is most commonly caused by iron deficiency [3]. Microcytic anemia is not uncommon in otherwise healthy people but with chronic anemia or those with genetic blood disorders such as thalassemia and sideroblastic anemia [3]. The peak at larger MCV values in Figure 4(b) was indicative of macrocytic anemia, that is, with MCV values above 95–100 fL, which is most commonly caused by vitamin B12 or folate deficiency [3]. The anemic samples were grouped into three categories on the basis of their RBC MCV values (microcytic with MCV values equal to and less than 82 fL, normocytic with MCV values above 82 fL and below 97 fL, and macrocytic with MCV values equal to and above 97 fL) and the summary of the RBC morphological parameters for these groups along with those for the samples from healthy donors is given in Table 5. It shows apparent morphological differences between the groups. The RBC morphological parameters of samples with normocytic anemia did not differ from those of the samples from healthy patients (P = 0.4 for length, P = 0.8 for width, P = 0.4 for surface area, P = 0.8 for sphericity, and P = 0.9 for aspect ratio, two-tail t-test for two samples with unequal variance) even though they were statistically different in MCHC (P < 0.001) and MCH (P = 0.005) values as noted above. Microcytic and macrocytic anemic samples had the same range of MCHC values as normocytic anemia samples (P = 0.8) but were different in morphology. Both microcytic and macrocytic RBCs were longer with mean length of 8.4 nm than normocytic RBCs with mean length of 8.0 nm (P < 0.001). Microcytic RBCs were also thinner with mean width of 1.7 nm than both normocytic and macrocytic RBCs with mean width of 2.2 nm and 2.1 nm, respectively (P = 0.004). Consequently, microcytic RBCs had lower sphericity and aspect ratio parameters than normocytic RBCs (P = 0.002). Macrocytic RBCs, in turn, had greater surface area than normocytic RBCs (P < 0.001). Severe anemia, that is, HGB < 8 g/dL, was not strongly associated with MCV and all three types of RBC morphology, microcytic, normocytic, and macrocytic, were observed in severe anemia samples. Yet, out of 21 samples with severe anemia, 15 were either microcytic or macrocytic (71%) whereas the majority of the samples with nonsevere anemia were normocytic (60%). The RBCs from samples with severe anemia had, on average, lower aspect ratio and sphericity parameters (P = 0.002). In this study, the RBC morphology could be associated with the causes of anemia and to some extent with its severity.


Characterization of red blood cells with multiwavelength transmission spectroscopy.

Serebrennikova YM, Huffman DE, Garcia-Rubio LH - Biomed Res Int (2015)

Histogram distribution plots of MCV (fL) index for samples from healthy donors (a) and anemic patients (b).
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4309305&req=5

fig4: Histogram distribution plots of MCV (fL) index for samples from healthy donors (a) and anemic patients (b).
Mentions: The MCV distribution for samples from healthy donors approached normal as can be seen in Figure 4(a). On the other hand, the distribution of the MCV values for the samples from anemic patients was more complex with side peaks at low and high MCV ranges (Figure 4(b)). Normocytic anemia, that is, with MCV values within 80–100 fL range, in the case of cancer patients is likely to be indicative of the bone marrow failure [3] and, depending on a variety of factors, including the type and intensity of chemotherapy, occurs in over 60% of cancer patients [18–20]. The bone marrow failure resulted in decreased production of functional RBCs and coupled with cancer related inflammatory response [18] and/or chemotherapy induced [19, 20] increase in destruction of RBCs; blood loss leads to decrease in HGB, RBC count, and HCT. Although hemolysis can also be associated with normocytic anemia, it was not detected in any sample of this study. The peak at smaller MCV values in Figure 4(b) was indicative of microcytic anemia, that is, with MCV values below 80 fL, which is most commonly caused by iron deficiency [3]. Microcytic anemia is not uncommon in otherwise healthy people but with chronic anemia or those with genetic blood disorders such as thalassemia and sideroblastic anemia [3]. The peak at larger MCV values in Figure 4(b) was indicative of macrocytic anemia, that is, with MCV values above 95–100 fL, which is most commonly caused by vitamin B12 or folate deficiency [3]. The anemic samples were grouped into three categories on the basis of their RBC MCV values (microcytic with MCV values equal to and less than 82 fL, normocytic with MCV values above 82 fL and below 97 fL, and macrocytic with MCV values equal to and above 97 fL) and the summary of the RBC morphological parameters for these groups along with those for the samples from healthy donors is given in Table 5. It shows apparent morphological differences between the groups. The RBC morphological parameters of samples with normocytic anemia did not differ from those of the samples from healthy patients (P = 0.4 for length, P = 0.8 for width, P = 0.4 for surface area, P = 0.8 for sphericity, and P = 0.9 for aspect ratio, two-tail t-test for two samples with unequal variance) even though they were statistically different in MCHC (P < 0.001) and MCH (P = 0.005) values as noted above. Microcytic and macrocytic anemic samples had the same range of MCHC values as normocytic anemia samples (P = 0.8) but were different in morphology. Both microcytic and macrocytic RBCs were longer with mean length of 8.4 nm than normocytic RBCs with mean length of 8.0 nm (P < 0.001). Microcytic RBCs were also thinner with mean width of 1.7 nm than both normocytic and macrocytic RBCs with mean width of 2.2 nm and 2.1 nm, respectively (P = 0.004). Consequently, microcytic RBCs had lower sphericity and aspect ratio parameters than normocytic RBCs (P = 0.002). Macrocytic RBCs, in turn, had greater surface area than normocytic RBCs (P < 0.001). Severe anemia, that is, HGB < 8 g/dL, was not strongly associated with MCV and all three types of RBC morphology, microcytic, normocytic, and macrocytic, were observed in severe anemia samples. Yet, out of 21 samples with severe anemia, 15 were either microcytic or macrocytic (71%) whereas the majority of the samples with nonsevere anemia were normocytic (60%). The RBCs from samples with severe anemia had, on average, lower aspect ratio and sphericity parameters (P = 0.002). In this study, the RBC morphology could be associated with the causes of anemia and to some extent with its severity.

Bottom Line: The MWT spectra were quantitatively analyzed with a Mie theory based interpretation model modified to incorporate the effects of the nonsphericity and orientation of RBCs.The changes in the RBC volume, surface area, aspect ratio, and hemoglobin composition were used to trace the morphological and compositional alterations in the infected RBCs occurring with parasites' development and to provide insights into parasite-host interactions.The MWT method was shown to be reliable for determination of the RBC morphological parameters and to be valuable for identification of the RBC pathologic changes and disease states.

View Article: PubMed Central - PubMed

Affiliation: Claro Scientific, LLC., 10100 Dr. Martin Luther King Jr. Street N., St. Petersburg, FL 33716, USA.

ABSTRACT
Multiwavelength transmission (MWT) spectroscopy was applied to the investigation of the morphological parameters and composition of red blood cells (RBCs). The MWT spectra were quantitatively analyzed with a Mie theory based interpretation model modified to incorporate the effects of the nonsphericity and orientation of RBCs. The MWT spectra of the healthy and anemic samples were investigated for the RBC indices in open and blinded studies. When MWT performance was evaluated against a standard reference system, very good agreement between two methods, with R (2) > 0.85 for all indices studied, was demonstrated. The RBC morphological parameters were used to characterize three types of anemia and to draw an association between RBC morphology and anemia severity. The MWT spectra of RBCs infected with malaria parasite Plasmodium falciparum at different life cycle stages were analyzed for RBC morphological parameters. The changes in the RBC volume, surface area, aspect ratio, and hemoglobin composition were used to trace the morphological and compositional alterations in the infected RBCs occurring with parasites' development and to provide insights into parasite-host interactions. The MWT method was shown to be reliable for determination of the RBC morphological parameters and to be valuable for identification of the RBC pathologic changes and disease states.

Show MeSH
Related in: MedlinePlus