doi.org/10.36721/PJPS.2020.33.2.REG.589-595.1 Pak. J. Pharm. Sci., Vol.33, No.2, March 2020, pp.589-595 589 Hematological complications under hyper conditions of glucose and distilled water Hafeez Ullah 1* , Munir Akhtar 2 , Fayyaz Hussain 2 , Ejaz Ahmad 2 , Mukhtar Ahmad 3 , Muhammad Hammad Aziz 3 , Farah Andleeb 4 and Zahida Batool 1 1 Bio-Photonics Research Laboratory Department of Physics, The Islamia University of Bahawalpur, Bahawalpur, Pakistan 2 Department of Physics, Bahauddin Zakariya University, University Campus, Bosan Road, Multan, Pakistan 3 Department of Physics, COMSATS Institute of Information and Technology, Lahore, Pakistan 1,4 Department of Physics, Govt. Sadiq College Women University, Bahawalpur, Pakistan Abstract: This work presents a pilot method of hematological diagnosis about changes in: shape, size and rouleaux formation, cell count of leucocytes and platelet cells in the presence of different glucose [C 6 H 12 O 6 ] and water [H 2 O] concentrations. The 2D microscopic images after addition of ten different glucose concentrations to normal blood (0 mM- 450 mM) revealed the lyses (disintegration) of white blood cells (WBCs). This work provides a baseline to diagnose blood disorders and complications at labs and clinical environment. Keywords: Glycolysis, hyperglycemia, glucometery, CBC (complete blood count). INTRODUCTION Blood is a fluid in our body which is not only responsible for transportation of nutrients within the body (proteins, fats, minerals and carbohydrates, etc.), but also waste materials (e.g. carbon-dioxide) out of body (Krieglstein et al., 2007). Usually 2-3 drops of blood contains 1 billion red blood cells (RBCs) and for every 600 RBCs there are only 1 WBC and 40 platelets (Ross et al., 1988). Normal glucose level after fasting is typically between 70 and 99mg/dL and 2 hours after eating a meal is less than 140mg/dL (Engelgau et al., 2000). Hyperglycemia is a condition where the blood contains more glucose than normal, and hypoglycemia, the blood contains less glucose than normal (Sperelakis, 2012). We have investigated three parameters of leucocytes i.e. (i) shape change, (ii) change in size and (iii) cell count under hyperglycemia and hypotonic solution of distilled water by using optical microscopy. The most prominent feature is that it does not use harmful ionizing radiation. In this work, we have used white light and dark field digital microscopy along with a CCD camera to record images in transmission mode. MATERIALS AND METHODS In this work, we have investigated two types of samples to diagnose blood parameters and blood cells. Sample set (I) Our sample set (I) consists of leucocytes with ten different concentrations (0mM, 50mM, 100mM, 150mM, 200mM, 250mM, 300mM, 350mM, 400mM and 450mM of glucose and distilled water. We took 10 samples of 2 ml of fresh blood for glucose and 4 ml for distilled water from healthy volunteer (blood type AB+) by vein puncturing. Samples were stored in Ethylenediaminetetra acetic acid (EDTA) tubes. We added the aforementioned concentrations of glucose and distilled water in each heparin tube 0 mM means normal blood i.e. no extra glucose or water addition. We used ethyl alcohol for fixing and field stain (A, B) for staining. Slides for each concentration were then examined using a white light microscope (Olympus CX41) at 100X after putting one drop of cedar wood emersion oil. This oil enhances the capability of lens. The resultant changes in shape and size of leucocytes were then captured using a CCD (Canon EOS 600D, Japan). The experiment was performed at room temperature (21 o C). Sample set (II) Our sample set (II) consists of platelets obtained by centrifuging the blood from same healthy male volunteer by vein puncturing. The blood was taken into ten Ethylenediaminetetraacetic acid (EDTA) anticoagulant tubes. Each tube contained 2 ml of blood for sugar and 4ml for water analyte. We mixed the nine different measured concentrations into nine different EDTA tubes. Ethical approval It is stated that all procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards. *Corresponding author: e-mail: hafeezullah79@gmail.com