145-0010
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Our lab uses insect cell cultures to produce recombinant mammalian proteins. For each protein that is produced using this method, considerable attention must be paid to the density of the cell cultures at various stages in the protocol. A failure to accurately document the density of cells in a culture will lead to a reduction in the efficiency of protein production or worse, complete cell death. When generating proteins using this method we often have 5-6 individual cell cultures incubating at a time. As a result, manual counting of cells in each culture can be needlessly time consuming. We bought the TC-10 to cut down the time spent counting cells and to standardize our protocols such that different users were able to count cells without user bias.
We use the TC-10 cell counter to (a) count and (b) determine viability of insect cells (Sf9 or High-Five) used for recombinant protein production. We use dual chamber disposable counting slides (Bio-Rad, #145-0011) in conjunction with the cell counter.
Cells in culture and a live/dead marker, e.g trypan blue.
To count insect cells, we remove our cultures from the incubator and transfer them to a tissue culture hood before removing 10ul of well-mixed culture in a sterile manner. This volume is mixed with an equal volume of trypan blue stain. This dye is absorbed by dead cells and is used to determine the viability of the cells under examination. When the slide and chamber are inserted into the instrument, the instrument automatically detects the presence of a slide and initiates a count. It will also automatically recognize that you have added trypan blue to the chamber and proceed to determine the viability of the cells. The instrument provides the number of cells/ml and the total number of live cells/ml – a number also displayed as a percentage (e.g. 98% live cells). Additionally, the user can view the cells at three different magnifications although the field of view cannot be changed. The instrument will automatically store previous counts which can be reviewed at a later date. Images associated with the counts are not stored. Cell counts and viability data can be exported via two USB ports located on the side of the instrument.
When insect cells are clumped together, the instrument determines them to be just one cell. As a result, it is worth viewing the cells on the screen to make sure that they are not clumped and therefore skewing the count. Additionally, if there is a sudden and unexpected drop in the viability of the cells, it is worth repeating the count. Occasionally, the focus function of the machine is slightly off, leading the instrument to interpret out-of-focus cells as being dead cells.
This instrument is extremely simple to use and accurate for use with insect cells. We have done several side by side comparisons with standard manual cell counting methods and results are consistent between the two platforms. The TC-10 is also accurate at determining the viability of insect cells in culture. Results, but not images, are readily exported via a USB connection.
Automatic slide detection, viability determination, visual inspection of cells at three magnifications, compact instrument, USB export, previous count storage
The TC-10 is capable of counting cells either with or without the addition of trypan blue (for viability determination). The machine is setup to automatically detect the presence of a dye added to the chamber in a slide.
Product Name: TC-10 Automated Cell CounterSupplier Page