Full spectrum of fluorescence detection—not limited to GFP and RFP
The Countess® II FL Automated Cell Counter has two fluorescence channels. The fluorescent colors collected are determined by the insertion of individual light cubes. With over 15 light cubes to choose from, the Countess® II FL Automated Cell Counter is not limited to detection of GFP- and RFP-expressing cells. Looking to measure the transfection efficiency of cells transduced with an RFP-expression construct with a blue nuclear counterstain? No problem. Simply add the RFP and DAPI light cubes, and you can effectively count the number of cells expressing the RFP fusion within the total population and accurately determine the transfection efficiency.
Two-color fluorescent cell counting.10 μL of HeLa cells transduced withCellLight® Histone 2B-RFPand stained withNucBlue® Live ReadyProbes® reagentwere added to achamber slideand inserted into a Countess® II FL Automated Cell Counter equipped withDAPIandRFPlight cubes. The segmentation and counting algorithms of the new Countess® II FL counter identifies and counts the cells based on their fluorescence properties. In this experiment, 86% of the HeLa cells within the population were transduced with the CellLight® Histone 2B-RFP construct.
EVOS® light cubes for the Countess® II FL Automated Cell Counter
Central to the fluorescence detection technology inside the Countess® II FL Counter are the proprietary EVOS® LED light cubes. Each cube contains an LED, illuminating optics, and filters. Light cubes are user interchangeable, auto-configured by the Countess® system with plug-and-play capability. The wide variety of light cubes available provides flexibility for multiple fluorescence research applications, such as cell viability analysis.
The mounting costs of disposable slides and tips have deterred many labs from giving up tedious manual cell counting. In response to this, the Countess® II FL Automated Cell Counter was designed to work with a reusable glass slide, which significantly reduces the long-term consumable costs of automated counting. For researchers who want the convenience of disposable slides, the Countess® II FL Automated Cell Counter can also use the same PMMA plastic slides developed for the original Countess® Cell Counter.
The Countess® II FL Automated Cell Counter contains advanced autofocusing and counting algorithms to allow you to quickly and accurately identify and count cells within a population. Upon insertion of the slide, the Countess® II FL counter will automatically focus on cells to help remove user variation associated with manual focusing mechanisms. Cells between 10 µm and 60 µm and in suspension at a density of 1 x 104to 1 x 107cells/mL are optimal for counting.
Precise cell counts with the Countess® II FL Automated Cell Counter.NIH/3T3, HeLa, and HL-60 cells were mixed 1:1 in suspension with a 0.4% solution ofTrypan Blue Stainand counted using the trypan blue exclusion method. A 10 μL sample was added to achamber slideand inserted into a Countess® II FL Automated Cell Counter (blue bar) or counted manually under a microscope using a hemocytometer (purple bar). Each bar is an average of 10 counts, with standard deviation as shown. The autofocusing and advanced counting algorithms of the Countess® II FL counter give more precise cell counts as compared with those derived from manual counting.
Not a problem with the Countess® II FL Automated Cell Counter. Whether you use automated or manual cell counting methods, obtaining accurate cell counts in samples containing clumpy cells is difficult to achieve. The advanced counting algorithms of the Countess® II FL Automated Cell Counter can identify clear delineations of cell boundaries within clumps of cells, thereby giving precise, accurate cell counts even with clumpy samples.
Accurate cell counts—even with clumpy samples.NIH/3T3 cells were mixed 1:1 in suspension with a 0.4% solution ofTrypan Blue Stain, and a 10 μL sample was added to achamber slideand inserted into a Countess® II FL Automated Cell Counter. The segmentation and counting algorithms of the Countess® II FL counter are able to resolve clumps of cells and score them as live or dead using the trypan blue exclusion method. In this sample, 3.09 x 106live cells (which excluded trypan blue stain) and 5.86 x 103dead cells (which accumulated trypan blue stain) were counted.
Updating software & registering your Countess® II or Countess® II FL
Learn how to update the software on your instrument. Also, use the button below to register your instrument.