Invitrogen™ Countess™ Cell Counting Chamber Slides and Holder, disposable

Catalog No.	Type	Quantity
T10282	Trypan Blue Stain	2 x 1 mL
C10228	Cell Counting Chamber Slide	50 Slides
C10312	Cell Counting Chamber Slide	500 Slides
C10313	Cell Counting Chamber Slide	1250 Slides
C10314	Cell Counting Chamber Slide	2500 Slides
C10315	Cell Counting Chamber Slide	5000 Slides
AMEP4745	Slide Holder	1 unit
C10283	Cell Counting Chamber Slide	50 Slides

Catalog No. T10282 Supplier Invitrogen™ Supplier No. T10282

Description

Includes

Sildes and 20 × 1mL vials of trypan blue (0.4%)

Countess Cell Counting Chamber Slides are conveniently packaged for use with the Countess Automated Cell Counter. Each disposable slide features two enclosed chambers. All Countess Cell Counting Chamber Slides (except Cat. No. C10283) are available with vials of trypan blue.

Countess Cell Counting Chamber Slides are designed exclusively for automated cell counting on the Countess Automated Cell Counter. These individually packaged slides feature two enclosed chambers and are disposable, offering convenience when performing cell culture. The Countess family of automated cell counters employs the widely used trypan blue staining method for cell counting to identify dead cells and measure cellular viability using brightfield.

The disposable Countess Cell Counting Chamber Slides offer convenience in automated cell counting for processing multiple cell samples, busy cell culture facilities, and easy disposal of contaminated or hazardous samples. They also eliminate the risk of contamination from residual samples. After use, Countess slides can be disposed of as standard biohazard waste. Unlike a hemocytometer, there is no need to clean Countess slides before or after use.

Each Countess Cell Counting Chamber Slide has two separate, enclosed chambers for counting one sample in duplicate or two different samples. Cell counting occurs in a central location of the counting chamber. The volume counted is 0.4 μL, the same as counting four 1-mm² squares in a standard hemocytometer.

The Countess Automated Cell Counter works in three simple steps:

• Mix 10 μL of sample with 10 μL of trypan blue stain or other desired reagent, including ReadyCount™ stains, and pipet into a disposable Countess chamber slide
• Insert the slide into the instrument
• Press 'Count'

That's it—get the total cell count, percent viable cells, and average cell size in as little as 30 seconds.

The Countess Cell Counting Chamber Slides include two vials of trypan blue for brightfield automated cell counting and viability analysis (Cat. Nos. C10228, C10312, C10314, C10313, and C10315). The slides can also be purchased as 1 box of 50 slides without trypan blue (Cat. No. C10283) for use with other stains such as the ReadyCount fluorescent stains. The trypan blue vials are also available for purchase without slides (Cat. No. T10282).

Trypan blue staining is a long-standing and widely used method to measure cellular viability by identifying dead cells. Live cells with intact membranes exclude the dye, while dead cells with compromised membranes become stained, allowing for the differentiation and counting of live and dead cells to assess cellular viability. Trypan blue staining is the method employed by the Countess family of automated cell counters for cell counting, and this 0.4% trypan blue solution is the correct concentration for use on these instruments.

Note: Precipitates are quite common in trypan blue, and typically increase with storage time and freezing. The three approaches below may help minimize trypan blue precipitate in your cell counting samples:

• Sterile filtration of trypan blue using 0.3-micron filter
• Centrifugation or passive settling (avoid mixing or vortexing trypan stock solutions)
• Gentle warming of the 1-mL trypan solution to 37°C for 10 minutes

The Countess Disposable Slide Holder is used to hold the Countess disposable slides. It is designed for use with Countess II FL, Countess 3, and Countess 3 FL automated cell counters and is included with these cell counters. A replacement Countess Disposable Slide Holder (Cat. No. AMEP4745 ) is available for separate purchase. This Countess Disposable Slide Holder is not compatible with the Countess Reusable Slide (Cat. No. A25750).

Specifications

• Type: Trypan Blue Stain
• For Use With (Equipment): Countess Automated Cell Counter
• Color: Blue
• Detection Method: Colorimetric
• Label or Dye: Trypan Blue
• Content And Storage: Trypan blue stain, 0.4% solution, 2 x 1 mL vials
  
  Store at room temperature.
• Quantity: 2 x 1 mL
• Shipping Condition: Room Temperature

Frequently Asked Questions (FAQs)

Why am I getting a much higher bead count on the Countess 3 Standard Slide (Cat. No. A51876)?

When counting cells that have been mixed with Trypan Blue, the Countess accounts for this 1:1 dilution and multiplies the count by 2.
When using the Countess 3 Standard Slide, the Trypan Blue correction box needs to be unchecked, since a 1:1 dilution was not made with Trypan Blue.

Is there a limit to the number of times I can warm up Trypan Blue Stain (0.4%) for use with the Countess Automated Cell Counter (Cat. No. T10282)? What is the importance of warming up in aliquots of 1 mL?

The Trypan Blue Stain (0.4%) can be warmed up to 37 degrees C numerous times. The entire 1 mL volume of the stain should be warmed up to ensure that you are getting the entire amount at the proper concentration of 0.4%; if you took out an aliquot to warm up, and left behind some precipitate, the aliquot that is warmed up may not contain the complete amount of dye and the final concentration may be less than 0.4%.

What is the shelf life of Trypan Blue Stain (0.4%) for use with the Countess Automated Cell Counter?

The shelf life of the product is 36 months from the date of production.

How long can cells be exposed to trypan blue before all cells are stained with the dye?

Unfortunately, there is no definitive answer to this question. The rate at which the cell-impermeant dye is absorbed depends on the cell type, their state of health, nourishment, engulfment activity, etc.

What causes trypan blue to form a precipitate?

Exposure to light may degrade the dye and these contaminants may promote precipitation. Trypan blue can also form orange/red fibrous aggregates if exposed to refrigeration or freezing temperatures.

How does trypan blue solution stain viable cells?

Trypan blue is a cell impermeant stain used to estimate the number of dead cells in a viable population. Its utility is based on the fact that it is a charged dye and does not enter cells unless the membrane is compromised. Live (viable) cells exclude the dye but dead (non-viable) cells or cells with a compromised membrane are stained an intense blue.

Prior to staining with trypan blue solution, do the cells need to be washed to remove the culture medium?

Trypan blue will bind to serum proteins as well as cellular proteins, which may result a high level of background staining. If the background is too dark, cells should be pelleted and resuspended in protein-free medium, buffer, or normal saline prior to counting.

Can I combine trypan blue staining with fluorescence staining?

It depends on the type of fluorescent stain used on the cells. Trypan blue is a cell-impermeant chromophore that can quench fluorescence. It may quench fluorescent staining on the surface of live cells or internal fluorescent staining in dead cells.

Can trypan blue solution be used to differentiate between apoptotic and necrotic cells?

Trypan blue will stain cells that have a compromised membrane. It cannot differentiate between compromised membranes caused by apoptosis or necrosis.

Do you have a protocol for cell counting via trypan blue exclusion?

Yes, you can find the protocol at the following link (https://www.thermofisher.com/us/en/home/references/gibco-cell-culture-basics/cell-culture-protocols/trypan-blue-exclusion.html)

How much Trypan Blue solution should I add to my cell suspension before counting?

We recommend adding 0.1 mL of 0.4% Trypan Blue stock solution to 0.1 mL of cells. However, depending upon the sample and instrumentation, the ratio of 0.4% Trypan Blue solution to the volume of cells can be varied from 1: 10 to 1:1 (v/v) so that the final concentration of Trypan Blue is 0.04% - 0.2%.

For Research Use Only. Not for use in diagnostic procedures.