Instructions: Need help with lab reports according to the given writing lab report writing manual. Experiment name is "Counting Cells" Table 1: Cell Counting Number of Living Cell (Unstained) Number of Dead Cell (Stained) Area 1 Area 2 Area 3 Area 4 Average Average number of living cells Average number of dead cells In order to fill this table, we have to count from the attached pictures./n İstanbul Bilgi Üniversitesi BIOE 222 CELL BIOLOGY LABORATORY Experiment 3 2023-2024 SPRING Dr. Tuğba KÖSE Ar. Gör. Betül BUDAK Page 1 EXPERIMENT 3 : COUNTING CELLS 0,100mm NEUBAJER Tiefe Depth Profondeur IMPROVED CE Optik abor 0,0025mm² Figure 1:Hemocytometer 1mm 1mm 1mm Identification of Dead and Living Cells Live cells contain intact cell membranes that eliminate certain dyes. In dead cells, the dye enters the cytoplasm and the cells take on the stain.(Figure 2) о O о Figure 3: Counting cell о Q Include Hemocytometer is a special type of microscope slide used to count blood cells and consists of two chambers, which is divided into nine (1.0mm x 1.0mm) large squares that are separated from one another by triple lines. The central counting area of the hemocytometer contains 25 large squares and each large square has 16 smaller squares. The area of each is 1mm² (Figure 1). C 60 0 0 CodonAug Figure 2: Live cells appear colourless and bright under phase contrast, while dead cells stain blue. cells touching middle on top and left Exclude cells touching middle line on bottom and right O ○ How do we count the cells? Counting cells on the hemocytometer can be confusing. You need to make sure you don't count the same cell twice. Otherwise, you will get the wrong cell count. You have to set a rule and strictly follow it. PROCEDURE Obtain a uniform suspension of cells: Suspension cell culture will be used. Place 100 microliters of cell suspension in a suitably-sized centrifuge tube. For an accurate cell count to be obtained, a uniform suspension containing single cells is necessary. Page 2 Prepare a 1:1 dilution of the cell suspension in trypan blue: Approximately 10 microliters of cell suspension will be required to charge one chamber of the hemocytometer. Combine the 100 microliters of cell suspension with the 100 microliters of trypan blue. Pipette up and down several times to ensure a uniform cell suspension. Prepare the counting chamber: The mirror-like polished surface is carefully cleaned with 70% ethanol and the cover slip is also cleaned. Moisten and affix cover slip to the hemocytometer. Load the hemocytometer: 10 microliters of trypan blue-cell suspension is transferred to one of the chambers of the hemocytometer by carefully touching the cover slip at its edge with the pipette tip and allowing each chamber to fill by capillary action. The chamber should not be overfilled or underfilled. Determine the number of cells (total and viable): View the cells under a microscope at 100x magnification. Under the microscope, focus on the corner squares. On each corner square, there are 16 small squares. You need to count the cells on each corner square (4X4). Keep a separate count of viable and non-viable cells. Please fill the Table 1. Table 1: Cell Counting Number of Living Cell (Unstained) Number of Dead Cell (Stained) Area 1 Area 2 Area 3 Area 4 Average Average number of living cells Average number of dead cells For your lab report you need to perform the following calculations. Calculation: % Cell Viability = [Total Viable cells (Unstained) / Total cells (Viable+Dead)] X 100 • Dilution Factor = *Total Volume / Volume of sample. *Total Volume= Volume of cell sample + Volume of diluting liquid (trypan blue) Viable Cells/ml = Viable cell count x Dilution Factor x 104 Page 3/n • Cover Page How to write? • Name of Experiment • Aim of Experiment • Introduction Materials and Methods • Results • Discussion 1 Table of Content Introduction......... • Materials & Method......... ● Results.......... 1 .3 ........7 2 TABLE OF FIGURES ? 3 TABLE OF FIGURES Figure 5.1: The name of the figure DO NOT PUT «FIGURES» ON THIS PAGE! 4 St TABLE OF FIGURES Figure 1.1: The name of the figure….... ● Figure 1.2: ……………………. ● Figure 3.1: !Each figure has to have a name!!! Figure 1.1: The name of the figure…………………………………..1 Number of Chapter Number of Figure for that Chapter .1 18 5

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Instructions: Need help with lab reports according to the given writing lab report writing manual. Experiment name is "Counting Cells" Table 1: Cell Counting Number of Living Cell (Unstained) Number of Dead Cell (Stained) Area 1 Area 2 Area 3 Area 4 Average Average number of living cells Average number of dead cells I