SALES BROCHURE. better count fast. easy and reliable cell counting. Countess Automated Cell Counter

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1 SALES BROCHURE better count fast easy and reliable cell counting Countess Automated Cell Counter

2 Consistent. Fast. Easy. The Countess Automated Cell Counter takes the subjectivity and tedium out of one of the fundamental steps of cell culture counting live and dead cells. By reducing user error and speeding up cell counting, the Countess Automated Cell Counter has revolutionized how researchers have counted cells with less sample and less hassle. Why use the Countess Automated Cell Counter? With the Countess Automated Cell Counter, producing quality reproducible data is fast and easy. It is also inexpensive and small, and can easily fit into a lab s budget and workspace. Accurate eliminate the subjectivity of manual cell counting; minimize guessing and user-to-user variability Fast counts live and dead cells, measures viability and average cell size in 30 seconds with just 10 μl of sample Convenient no setup, cleaning, or service required Adaptable optimize counting protocols for your specific cell line Reliable gating allows counting of specific cell types in mixed populations The small footprint (27 cm (w) x 20 cm (d) x 19 cm (h)), and data archiving function of the Countess Automated Cell Counter make sharing between labs and users easy. In addition, the disposable cell counting chamber slides minimize exposure of users to biohazardous samples. The Countess Automated Cell Counter is ideal for: Cell culture labs Flow cytometry core facilities HTS labs HIV and other infectious disease labs Stem cell research labs The Countess Automated Cell Counter is easy to use (Figure 1). Simply pipet the sample into the counting slide (A), insert the slide into the Countess Automated Cell Counter (B), then press Count cells (C); results are displayed in 30 seconds (D). A B C D Figure 1. The Countess Automated Cell Counter workflow. Pipet the sample into the counting slide (A); insert the slide into the Countess Automated Cell Counter (B); press Count cells (C); results are displayed in 30 seconds (D).

3 Measured concentration (cells/ml) How does the Countess Automated Cell Counter work? The Countess Automated Cell Counter uses trypan blue staining combined with a sophisticated image analysis algorithm to produce accurate cell and viability counts in as little as 30 seconds Countess Automated Cell Counter Hemocytometer The algorithm also measures the average size of live, dead, and total cells, to give you all the data you need to proceed with your experiments. The measurement range extends from 1 x 10 4 to 1 x 10 7 cells/ml, with an optimal range from 1 x 10 5 to 4 x 10 6 cells/ml, broader than that of a hemocytometer (Figure 2). The optimal cell size is between 5 μm and 60 μm. For accurate viability counts, ensure the counting area is covered with cell suspension and count cells within 2 minutes of mixing the cells with trypan blue solution. We highly recommend that you use the default setting to begin with for your cell counting, unless you know the default setting is not suited to your particular cell sample. For the best data with biological samples, we recommend counting at least two samples and taking an average. A handy dilution calculator helps you determine how to prepare your sample for your next passage or experiment. Adjust the focus knob to optimize the image for analysis as follows (Figure 3): Live cells have bright centers and dark edges Dead cells have a uniform blue color throughout the cell with no bright centers Cell suspension dilution (cells/ml) Figure 2. Data from the Countess Automated Cell Counter extend further into the high concentration range than hemocytometer readings. A B Dead Live Dead Live C Dead Live Figure 3. Correct and incorrect images. (A) Correct image: live cells have bright centers and dark edges. (B) Incorrect image: dead cells have bright, blue centers and are counted as live. (C) Incorrect image: live cells have dark centers and are counted as dead.

4 How accurate is the Countess Automated Cell Counter? The Countess Automated Cell Counter was compared with the hemocytometer and Coulter Cell and Particle Counter methods (Table 1, Figures 4 and 5). Download the complete technical notes including protocols and cell counting tips at Table 1. Comparison between the Countess Automated Cell Counter and Coulter Cell and Particle Counter method of cell counting. Feature Countess Automated Cell Counter Coulter Cell and Particle Counter Viability data Provided None provided Cell size data Provided None provided Results visualization Provided None provided Setup Easy Difficult Calibration/maintenance None required Required daily/monthly 1.6 x x x x x x 10 6 Cells/mL 1.0 x x x 10 4 Beads/mL 8 x x x x x x Z1 Series Coulter Counter Countess Automated Cell Counter Cell counting method 0 Countess Automated Cell Counter Glass hemocytometer Disposable hemocytometer Cell counting method Bead standard Figure 4. Equivalent results between Countess Automated Cell Counter and Coulter Cell and Particle Counter methods of cell counting. K562 cells were counted using the Z1 Series Coulter Counter Cell and Particle Counter and the Countess Automated Cell Counter. Cell counts from replicate samples were obtained on the Countess Automated Cell Counter according to the manufacturer s instructions. Replicate samples were prepared for analysis on the Coulter Cell and Particle Counter instrument by diluting 100 μl of K562 cells into 10 ml of Isoton II Diluent. Figure 5. Accuracy and precision of the Countess Automated Cell Counter compared to glass and disposable hemocytometer methods. A standard bead solution of known concentration (9.57 x 10 5 beads/ml, Beckman Coulter) was used as the starting material for counting measurements using the Countess Automated Cell Counter, Bright-Line glass hemocytometer (Hausser Scientific), and C-Chip disposable hemocytometer (Digital Bio). The beads were mixed 1:1 with 0.4% trypan blue, and triplicate samples were measured according to each manufacturer s instructions. Accuracy was evaluated by comparing the mean concentration (in cells/ml) measured by each method to the expected concentration of the standardized bead solution x 10 5 ± 10% beads/ml). Precision is indicated by the standard deviations; error bars represent one standard deviation. Accuracy is comparable between the Countess Automated Cell Counter and the disposable hemocytometer, and the Countess Automated Cell Counter is more precise than the hemocytometers.

5 What can be counted with the Countess Automated Cell Counter? Table 2 lists some of the cell lines validated on the Countess Automated Cell Counter. Some other counting capabilities include: White blood cells The Countess Automated Cell Counter is able to count white blood cells from lysed whole blood and Ficoll cell preparations. Whole blood cells containing non-lysed cells Dilute the blood sample approximately 1:10,000 and count in bead mode. The Countess Automated Cell Counter cannot assess the viability of cells in a whole blood sample. PBMCs (peripheral blood mononuclear cells) The Countess Automated Cell Counter can count PBMCs. However, it cannot differentiate white blood cell types. RBCs (red blood cells) The Countess Automated Cell Counter can count RBCs. Dilute the blood sample approximately 1:10,000 and count in bead mode. The instrument cannot assess the viability of red blood cells. In addition, the instrument cannot distinguish red blood cells from white blood cells. Read about how to count blood cells and download this application note at Bookmark this page, as additional application notes are added regularly. The Countess Automated Cell Counter can accurately count clumps of cells. In some cases, the actual number of cells in a clump will be underestimated and the cell size will sometimes be overestimated. However, we have found that the Countess Automated Cell Counter provides data at least as accurate as counting with a hemocytometer. Yeast cells We have successfully counted Saccharomyces cerevisiae (Fleischmann s baker s yeast), a consumer product. However, the instrument cannot distinguish yeast viability. Plant cells Plant cells have not been tested. If the plant cells fall within the optimal size range for the instrument and are not extremely clumpy, the instrument should be able to count plant cells. Elongated cells We have not tested very irregularly shaped or elongated cells. If the cells are very irregular or elongated, try the Parameters function under the Settings menu to vary the circularity. This function alters the way that the image analysis software recognizes cells. You may need to experiment with several circularity settings until you find the one that is perfect for your cell type. Bacterial cells Bacteria are too small to be distinguished from non-cell debris in the Countess Automated Cell Counter. Sperm and other fast-moving cells It is difficult to count cells that are moving quickly. The Countess Automated Cell Counter does not count live sperm and fast-moving protozoa. Hepatocytes The instrument is able to calculate total concentration, but cannot provide viability information on the cells. Set the instrument to bead mode, then you can perform the cell counting.

6 Table 2. Cell lines validated on the Countess Automated Cell Counter. For more information on our findings, visit Cell type Vendor Cat. No. Animal Organ Cell size (diameter) A431 ATCC CRL-2592 Human Skin 15.5 μm ADSC Invitrogen R Human Adipose-derive stem cells 13 μm Aortic smooth muscle Invitrogen C-007-5C Human Smooth muscle 20 μm Blood, whole lysed Donor NA Human Blood NA CHO-M1WT2 ATCC CRL-1984 Chinese hamster Ovary NA CHSE ATCC CRL-1861 Chinook salmon Embryo μm COLO-205 ATCC CCL-243 Human Colon NA COS-7 ATCC CRK-1651 African monkey Kidney NA ESC Millipore CMTI-1 Mouse Embryo 10 μm HEK-293 ATCC CRL-1573 Human Kidney 13 μm HEKn Invitrogen C-001-5C Human Neonatal foreskin 12.5 μm HeLa ATCC CCL-2 Human Cervix NA HepG2 ATCC CRL Human Liver 18 μm HESC WiCell NA Human Enbryo μm HL-60 ATCC CCL-240 Human Blood NA J774A.1 ATCC TIB-67 Mouse Blood μm Jurkat ATCC TIB-152 Human Blood 12 μm K-562 ATCC CCL-243 Human Bone marrow NA MCF7 ATCC HTB-22 Human Breast μm MRC-5 ATCC CCL-171 Human Lung 18 μm NIH/3T3 ATCC CRL-1658 Human Embryo 18 μm NSC Invitrogen N Rat Brain 11 μm PC-12 ATCC CRL-1721 Rat Adrenal gland μm Pulmonary artery endothelial Invitrogen C-008-5C Human Blood vessel 13 μm Pulmonary artery smooth muscle Invitrogen C-009-5C Human Smooth muscle 20 μm SF-21 Invitrogen Insect Ovary NA U266 ATCC TIB-196 Human Blood μm U-2 OS ATCC HTB-96 Human Bone NA Umbilical vein endothelial Invitrogen C-015-5C Human Blood vessel 17 μm

7 Has the Countess Automated Cell Counter been referenced in peer-reviewed publications? The Countess Automated Counter has been referenced in over 100 publications using over 120 different cell types from 18 different species. For a complete list of references, visit References Bovine chondrocytes, primary Mhanna RF et al. (2011) Layer-by-Layer Films Made from Extracellular Matrix Macromolecules on Silicone Substrates. Biomacromolecules. 12: Canine MDCK kidney epithelial cells Kakiashvili E et al. (2011) The epidermal growth factor receptor mediates Tumor Necrosis Factor-α induced activation of the ERK/GEF-H1/ RhoA pathway in tubular epithelium. JBC. 286: Dictyostelium discoidum, amoebae Fumanelli L et al. (2011) Mathematical modeling of bacterial virulence and host pathogen interactions in the Dictyostelium/Pseudomonas system. J Theor. Bio. 270: Human A549 Adenocarcinomic alveolar basal epithelial cells García-Álvarez I et al. (2011) Lipid and ganglioside alterations in tumor cells treated with antimitotic oleyl glycoside. Mol. Biosys. 7: Human colorectal carcinoma cells Williams BS et al. (2010) Mediated Decay Resistant Mutations Are a Source of Expressed Mutant Proteins in Colon Cancer Cell Lines with Microsatellite Instability. PLoS ONE doi: /journal.pone Human DU145 prostate cancer cells Chen H-M et al. (2011) A novel synthetic protoapigenone analogue, WYC02-9, induces DNA damage and apoptosis in DU145 prostate cancer cells through generation of reactive oxygen species. Free Rad Bio Med. 50: Human EOL-1 eosinophilic leukemic cell line Kahn J-E et al. (2011) Comparative Proteomic Analysis of Blood Eosinophils Reveals Redox Signaling Modifications in Patients with FIP1L1-PDGFRA-Associated Chronic Eosinophilic Leukemia. J. Proteome Res. 10: Human embryonic stem cells Wilson KD et al. (2010) Effects of Ionizing Radiation on Self-Renewal and Pluripotency of Human Embryonic Stem Cells. Cancer Res. 70: Human fibroblasts, primary Churko JM et al. (2011) Human dermal fibroblasts derived from oculodentodigital dysplasia patients suggest that patients may have wound-healing defects. Human Maturation. 32: Human H1299 lung carcinoma cell line Chen JY et al. (2010) Additive effects of C2-ceramide on paclitaxelinduced premature senescence of human lung cancer cells. Life Sciences 87: Human mesenchymal stem cells Genetos DC (2010) Betacellulin inhibits osteogenic differentiation and stimulates proliferation through HIF-1α. Cell Tissue Res 340: Human neuroblastoma cell lines Brignole C et al. (2010) Therapeutic targeting of TLR9 inhibits cell growth and induces apoptosis in neuroblastoma. Cancer Res. 70: Human whole blood Koutna I et al. (2011) Proliferation and differentiation potential of CD133+ and CD34+ populations from the bone marrow and mobilized peripheral blood. Ann Hematol 90: Human U-2 osteosarcoma cell line Beacham DW et al. (2010) Cell-based potassium ion channel screening using the FluxOR assay. J Biomol Screen. 15: Human U251 glioblastoma cells Aka A (2010) Label-Free Determination of the Number of Biomolecules Attached to Cells by Measurement of the Cell s Electrophoretic Mobility in a Microchannel. PLoS ONE 5(12): e Human WI-38T embryonic lung fibroblasts Buganim Y et al (2010) A Novel Translocation Breakpoint within the BPTF Gene Is Associated with a Pre-Malignant Phenotype. PLoS ONE 5(3): e9657. Mouse Ba/F3 pro-b cells Wasag B et al (2011) The kinase inhibitor TKI258 is active against the novel CUX1-FGFR1 fusion detected in a patient with T-lymphoblastic leukemia/lymphoma and t(7;8)(q22;p11) Haematologica / haematol Mouse C2C12 myoblasts, human mesenchymal stem cells, bovine chondrocytes Guillaume-Gentil O et al. (2011) Electrochemically switchable platform for the micro-patterning and release of heterotypic cell sheets. Biomed Microdevices 13: Mouse embryonic cardiomyocytes, primary Lai D et al. (2010) Neuregulin 1 sustains the gene regulatory network in both trabecular and nontrabecular myocardium. Circ. Res. 107: Mouse induced pluripotent stem cells (ipsc) Polo JM, et al. (2010) Cell type of origin influences the molecular and functional properties of mouse induced pluripotent stem cells. Nat. Biotech. 28: Mouse melanocytes, primary Mishra PJ et al. (2010) Dissection of RAS downstream pathways in melanomagenesis: a role for Ral in transformation. Oncogene 29: Rainbow trout enterocytes Kwong RWM et al (2010) Molecular evidence and physiological characterization of iron absorption in isolated enterocytes of rainbow trout (Oncorhynchus mykiss): Implications for dietary cadmium and lead absorption. Aquatic Toxicology. 99: Rat INS-1 832/13 pancreatic cells Altirribe J et al. (2010) The role of transmembrane protein 27 (TMEM27) in islet physiology and its potential use as a beta cell mass biomarker. Diabetologia 53: Rat olfactory ensheathing cells Dai C et al. (2010) Survival of retinal ganglion cells in slice culture provides a rapid screen for olfactory ensheathing cell preparations. Brain Research. 1354: Rat PC12 adrenal pheochromocytoma cell line Maheshwari M et al. (2010) Hydralazine Modifies Aβ Fibril Formation and Prevents Modification by Lipids in Vitro. Biochemistry. 49: Zebrafish ZEB2J blastula cells Monaghan SR et al. (2011) In vitro growth of microsporidia Anncaliia algerae in cell lines from warm water fish. In Vitro Cell Dev Biol Anim 47:

8 See what Countess users have to say: The Countess Automated Cell Counter makes tissue culture work much easier and more productive. I wish I had one years ago! John McGrath, Dana-Farber Cancer Institute I am using the Countess Automated Cell Counter to verify cell count and viability following cell sorting by flow cytometry. I have found the output to be very useful as verification of a successful sort. Users have found the output to be a time-saving tool as they prepare their cells for downstream processing. S. Schloemann, Washington University The Countess Automated Cell Counter is very easy to set up and use. Great cell counting instrument for a small lab or a large group of users. No mess left behind when counting is done, unlike Coulter Counter where you have to blank after each use. Alexandra Lin, National Institutes of Health The Countess Automated Cell Counter has worked out well for our lab. The Countess intrument cell counts agree with our manual cell counts, and the Countess Automated Cell Counter is much faster. Danielle Krebs, UBC Life Sciences Centre We found the Countess Automated Cell Counter very helpful for our migration assays. Instead of spending hours counting cells to get results, we are able to quickly quantify our data! A definite time saver and well worth the cost! Holly, University of Rochester The Countess Automated Cell Counter has proven to be a huge time saver. It increases productivity at all levels. John Coburn, Massachusetts General Hospital The Countess Automated Cell Counter saves us hours of time during experimentally intense workdays. We also appreciate the consistency of counts even with different users. Sarah, University of Illinois My experiences thus far have been pleasant. The Countess Automated Cell Counter rivals the precision of my Coulter Counter. Peter, Louisiana State University Read more customer commentary at Ordering information Description Quantity Cat. No. Countess Automated Cell Counter each C10227 Countess Automated Cell Counter Starter Kit 1 kit C10310 (includes 1 cell counter and 11 boxes of slides) Countess Automated Cell Counter Lab Starter Kit 1 kit C10311 (includes 1 cell counter and 101 boxes of slides) Countess Cell Counting Chamber Slides 50 slides (100 counts) C slides (1,000 counts) C ,250 slides (2,500 counts) C ,500 slides (5,000 counts) C ,000 slides (10,000 counts) C10315 Life Technologies offers a breadth of products DNA RNA PROTEIN CELL CULTURE INSTRUMENTS For Research Use Only. Not intended for any animal or human therapeutic or diagnostic use Life Technologies Corporation. All rights reserved. The trademarks mentioned herein are the property of Life Technologies Corporation or their respective owners. Printed in the USA. CO Headquarters 5791 Van Allen Way Carlsbad, CA USA Phone Toll Free in the USA