Learn the fundamentals of fluorescence imaging.
Countess® II FL Automated Cell Counter
Fast, easy, affordable cell counting
With fluorescence capabilities—bright-field and two optional fluorescence channels—the Countess® II FL can count cells, monitor fluorescent protein expression, and measure cell viability.
Watch the video: Using the Countess® II FL Counter
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 with CellLight® Histone 2B-RFP and stained with NucBlue® Live ReadyProbes® reagent were added to a chamber slide and inserted into a Countess® II FL Automated Cell Counter equipped with DAPI and RFP light 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.|
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.
- See the entire line of compatible EVOS® light cubes
- Watch the video: How Simple it is to Change EVOS Light Cubes
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 104 to 1 x 107 cells/mL are optimal for counting.
Download the application note: Fluorescent viability assays on the Countess® II FL Automated Cell Counter ›
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 of Trypan Blue Stain and counted using the trypan blue exclusion method. A 10 μL sample was added to a chamber slide and 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.
|How to focus||Performing a simple brightfield count|
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 of Trypan Blue Stain, and a 10 μL sample was added to a chamber slide and 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 106 live cells (which excluded trypan blue stain) and 5.86 x 103 dead cells (which accumulated trypan blue stain) were counted.
|Feature||Countess® II||Countess® II FL
|Upgradable to fluorescence|
|Reusable analysis slide
|Disposable analysis slide||
|Included in the box||
|Adipocytes||Human||Adipose-derived stem cells|
|Aortic Smooth Muscle||Human||Smooth muscle|
|Pulmonary artery endothelial||Human||Blood vessel|
|Pulmonary artery smooth muscle||Human||Smooth muscle|
|Umbilical vein endothelial||Human||Blood vessel|
|Whole lysed blood||Human||Blood|
Countess® II/II FL software download
Original Countess® software download
Comprehensive guide to fluorescent probes and labeling technologies
Compare multiple dyes and design and select a combination of cell structure probes.