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)
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O
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Figure 3: Counting cell
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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
