Correlation of Microscopic and Automated Cell Counter Results of Newer Hematological Variables in Diagnosis and Recovery of Anemia

  • Ahuja Ankur Army Hospital (Research & Referral) Delhi, India
  • Pandey Pankaj All India Institute of Medical Sciences, Rishikesh, Uttarankhand, India
  • Gahlot Gaurav PS Army Hospital (Research & Referral) Delhi, India
  • Badwal Sonia Military Hospital Shimla, Himachal Pradesh, India
  • Moond Saurabh Vardhman Mahavir Medical College & Safdarjung Hospital, Delhi, India
  • Dhanetwal Murari Lal Jaipur National University Institute for Medical Sciences and Research Centre, Jaipur India
Keywords: Automated cell counters, Iron deficiency anemia, thalassemia, Reticulocyte count, Immature reticulocyte fraction, Mean reticulocyte volume, Mean reticulocyte hemoglobin content, Fragmented RBC count


Background:  The correct interpretation of new hematological parameters generated by Automated Cell Counters (ACCs) requires extensive knowledge for the clinical significance of these results. The present study will enlighten the vital role of the traditional & new parameters of the Complete Blood Count (CBC) in early diagnosis and follow up stage after treatment especially in the setting of anemia Aims:  To evaluate the vast & extended utility of newer hematological parameters generated by ACCs beyond their conventional role of CBC and Differential Leukocyte Count (DLC) Study design: This is a cross sectional observational study of results obtained from blood samples of 100 patients having various types of anemia diagnosed over a period of 18 months Subjects & methods: EDTA blood samples of the patient were collected under aseptic precautions and processed within 2 hours through new generation hematology analyzer i.e. ADVIAR 2120i. The peripheral blood film examination in each case was performed on Leishman-Geimsa stained smear. Second EDTA samples were taken on 2nd and 10th day of follow up of IDA and thalassemia cases respectively. Statistical Analysis Used: The hematological, parameters obtained from ACCs/microscopic were assessed by Student’s t-test, Fisher’s exact test using the SPSS 21.0 software with considering p value < 0.05 as significant Results:  IRF and reticulocyte% can distinguish well between IDA and thalassemia major. CHr and MCVr are useful parameter for diagnosis of IDA/thalassemia and vitamin B12 deficiency anemia. CHr is the first parameter for monitoring response treatment of anemia.

Author Biographies

Ahuja Ankur, Army Hospital (Research & Referral) Delhi, India
Dept of Lab & Mol Med
Pandey Pankaj, All India Institute of Medical Sciences, Rishikesh, Uttarankhand, India
Dept of Pathology
Gahlot Gaurav PS, Army Hospital (Research & Referral) Delhi, India
Dept of Lab & Mol Med
Badwal Sonia, Military Hospital Shimla, Himachal Pradesh, India
Commanding Officer
Moond Saurabh, Vardhman Mahavir Medical College & Safdarjung Hospital, Delhi, India
Dept of Pathology
Dhanetwal Murari Lal, Jaipur National University Institute for Medical Sciences and Research Centre, Jaipur India
Dept of Pathology


1. Vierordt K. New method of quantitative microscopic analysis of the blood. Arch of
physiol. 1852; 41: 356-60. [2]
2. Coulter WH. Means for counting particles suspended in a fluid. 1953; 10: 508-13. [7]
3. Lesesve, J Martin, M Banasiak, C Andre-Kerneis, E Bardet, V Lusina, D
Kharbach, A Genevieve, F Lecompte. Schistocytes in disseminated intravascular
coagulation. International Journal of Laboratory Hematology. 2014; 36: 439-443. [73]
4. Zini G, d’Onofrio G, Briggs C, et al. ICSH recommendations for identification,
diagnostic value, and quantitation of schistocytes. Int J Lab Hematol. 2012; 34: 107-16. [168]
5. Yesmin MS, Sultana T, Roy CK, Rehman MQ, Ahmed ANN. Immature reticulocyte
fraction as a predictor of bone marrow recovery in children with acute lymphoblastic
leukemia on remission induction phase. Bangladesh Med Counc Bull. 2011; 37: 57-60. [80]
6. Advani SH, Giri NK. Acute lymphoblastic leukemia in childhood : treatment, results and
prognostic factors. Indian journal cancer. 1989; 26: 180-8. [87]
7. Kabata J, Tichelli A, Gratwohl A, Speck B. Flow cytometric pat¬tern of leucocyte
recovery after therapy induced aplasia. Acta Hematol Pol. 1994; 25: 329-42. [81]
8. Briggs C, Bain BJ. Basic haematological techniques. Dacie and Lewis Practical
Haematology. 2012; 11: 342-58. [131]
9. Mast AE, Blinder MA, Lu Q, et al. Clinical utility of the reticulocyte hemoglobin
content in the diagnosis of iron deficiency. Blood. 2002; 99: 1489-1491. [138]
10. Khusun H, Yip R, Schultink W, Dillon DH. World Health Organization hemoglobin cut-off points for the detection of anemia are valid for an Indonesian population. J Nutr. 1999;129:1669-74.

11. Zakai NA, French B, Arnold AM, Newman AB, Fried LF, Robbins J, Chaves P, Cushman M. Hemoglobin decline, function, and mortality in the elderly: the cardiovascular health study. Am J Hematol. 2013;88:5-9.

12. Gulati G, Behling E, Kocher W, Schwarting R. An evaluation of the performance of
Sysmex XE-2100 in enumerating nucleated red cells in peripheral blood. Arch Pathol
Lab Med. 2007; 131: 1077-83. [166]
13. Cui W, Wu W, Wang X, et al. Development of the personalized criteria for microscopic review following four different series of hematology analyzer in a Chinese large scale hospital. Chin Med J. 2010; 123: 3231-7. [167]
14. J-F Lesesve, V Asnafi, F Braun, G Zini. Fragmented red blood cells automated
measurement is a useful parameter to exclude schistocytes on the blood film. International Journal of Laboratory Hematology. 2012; 34: 566-76. [169]
15. Jean-Francois Lesesve, Amandine Daigney, Sylvain Henry, Elodie Speyer. Fragmented
red cells reference range (Sysmex XN(®) automated blood cell counter). Annales de Biologie Clinique. 2015; 73: 547-56. [170]
16. Łuczyński W, Ratomski K, Wysocka J , Krawczuk-Rybak M , Jankiewicz J. Immature
reticulocyte fraction (IRF) – an universal markerof hemopoiesis in children with cancer?
Eur J Haematol. 1994; 53: 293-7. [171]
17. Buttarello M, Temporin V, Ceravolo R, Farina G, Bulian P. The new reticulocyte
parameter (Ret-y) of the Sysmex XE 2100. Its use in the diagnosis and monitoring of
post treatment sideropenic anemia. Am J Clin Pathol. 2004; 121: 489-95. [178]
18. C. Ceylan, M. Miskiog, H. C. Olak, B. Kilic¸ E. O. Zdemir. Evaluation of reticulocyte
parameters in iron deficiency, vitamin b12 deficiency and b-thalassemia patients. Int J Lab Hematol. 2007; 29: 327-334. [181]
19. McClatchey KD. Clinical laboratory medicine. Philadelphia Lippincott Willimas and
Wilkins. 2002; 8: 609-20. [184]
20. d’Onofrio G, Chirillo R, Zini G, Caenaro G, Tommasi M, Micciulli G. Simultaneous
measurement of reticulocyte and red cell indices in healthy subjects and patients with microcytic and microcytic anemia. Blood. 1995; 85: 818-23. [182]
Original Article