i am working on a project where i need to find a protocol that shows how to quantify cultured cells inside a stirred tank bioreactor but every where i searched i found nothing
You could measure the increment of the cell number or the increment in the biomass of the cells.
Methods measuring the number of cells: Manual microscopy, automatized microscopy, Coulter counter, viable count (if you are working with microorganisms).
Methods measuring the biomass: Dry-weight, wet-weight, turbidimetry.
Methods measuring components of the cell: Protein, DNA, ATP, fatty acids.
Methods measuring products of cellular metabolism: Organic carbon (carbon dioxide), electric conductivity.
All methods have pros and cons and should be selected based on what kind of cells you are working with and the characteristics of the culture media (viscosity, density, turbidity). Also, many Bioreactor fabricants sell probes customized for different purposes, and one of the mentioned methods should be the fundament of the measuring.
I hope I've helped.
I do not know where you searched cuz that's an easy one, most common of the tasks in microbiology and related fields.
The best method depends on several factors, including:
- Cell type: Different cell types have different characteristics that may influence the chosen method.
- Culture conditions: Factors like cell density and media composition can affect the suitability of specific methods.
- Desired information: Are you interested in total cell number, viable cell count, or another parameter?
Here are some common methods for quantifying cells in stirred tank bioreactors:
Direct cell counting:
- Hemocytometer: This is a classic method using a microscope and a specialized counting chamber. It's most suitable for low-density cultures and requires aseptic technique for sampling.
- Automated cell counters: These instruments use various principles like electrical impedance or fluorescence to count cells. They are faster than hemocytometers but may require specific sample preparation and expensive equipment.
Indirect methods:
- Turbidity: Cell density can be estimated by measuring the culture's optical density (OD) at a specific wavelength, usually 600nm. This method is simple but not very accurate for dense cultures or cultures with debris.
- Biomass measurement: This involves drying or filtering a sample to determine the total cell mass. It's not specific to cells but can be useful for monitoring overall growth.
- Metabolic activity assays: These assays measure cellular activity by quantifying products like ATP or lactate. They don't directly measure cell number but can be indicative of viable cell mass.
Next time, you might need to start by reading your Microbiology Textbook
what type of this culture? what type of reactant ? these things determined the quantity of culture.
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