Chapter 3S
Viable Cell Enumeration
Cell counting is necessary not only to obtain data on the growth curve of
a culture, but also to determine the cell density of a sample to be used in
experiments. There are several methods for counting cell numbers.
Subprotocol 1
Hemocytometer Count
• • Materials
- Hemocytometer (Thoma, Neubauer, or Biirker-TUrk; Fig. 1)
- Biologic microscope
- Pasteur pipettes
- Micropipettes
- Hand-held tally counter
- Physiological solution
- Culture media
- 0.3% Trypan blue in physiologic solution or culture media
Burker-Turk type (depth: 0.1 mm)
Thoma type (depth: 0.1 mm)
Improved Neubauer type
(depth: 0.1 mm)
TATAI type (depth: 0.2 mm)
Fig. 1. Various types of hemocytometers Reproduced from Onishi K. (1996) with
permission of Asakura Shoten
PROTOCOL
Equipment
Solutions
Viable Cell Enumeration
Cell counting is necessary not only to obtain data on the growth curve of
a culture, but also to determine the cell density of a sample to be used in
experiments. There are several methods for counting cell numbers.
Subprotocol 1
Hemocytometer Count
• • Materials
- Hemocytometer (Thoma, Neubauer, or Biirker-TUrk; Fig. 1)
- Biologic microscope
- Pasteur pipettes
- Micropipettes
- Hand-held tally counter
- Physiological solution
- Culture media
- 0.3% Trypan blue in physiologic solution or culture media
Burker-Turk type (depth: 0.1 mm)
Thoma type (depth: 0.1 mm)
Improved Neubauer type
(depth: 0.1 mm)
TATAI type (depth: 0.2 mm)
Fig. 1. Various types of hemocytometers Reproduced from Onishi K. (1996) with
permission of Asakura Shoten
PROTOCOL
Equipment
Solutions